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1. Muscular Dystrophy Mutations Impair the Nuclear Envelope Emerin Self-assembly Properties

Author: Brigitte Buendia, Dmytro Puchkov, Sophie Zinn-Justin, Christophe Velours, Camille Samson, Louis Renault, Howard J. Worman, Pierre Chervy, Cecilia Östlund, Isaline Herrada, Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Department of Medicine, Columbia University [New York]-College of Physicians and Surgeons, Department of Pathology and Cell Biology, Department of Molecular Pharmacology and Cell Biology, Leibniz Institute of Molecular Pharmacology, FMP Berlin, Unité de Biologie Fonctionnelle et Adaptative (BFA (UMR_8251 / U1133)), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Leibniz Forschungsinstitut für Molekulare Pharmakolgie = Leibniz Institute for Molecular Pharmacology [Berlin, Allemagne] (FMP), Leibniz Association-Leibniz Association, Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Biologie Fonctionnelle et Adaptative ( BFA ), and Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS )
Subjects: Magnetic Resonance Spectroscopy, Nuclear Envelope, [SDV]Life Sciences [q-bio], Emerin, Biochemistry, Muscular Dystrophies, Article, Frameshift mutation, Spectroscopy, Fourier Transform Infrared, medicine, Humans, Missense mutation, Proteostasis Deficiencies, Muscular dystrophy, Nuclear protein, Gene, Loss function, [ SDV ] Life Sciences [q-bio], Chemistry, Genetic Variation, Membrane Proteins, Nuclear Proteins, General Medicine, medicine.disease, Cell biology, Membrane protein, Molecular Medicine, Hydrophobic and Hydrophilic Interactions, HeLa Cells
Abstract: International audience; More than 100 genetic mutations causing X-linked Emery-Dreifuss muscular dystrophy have been identified in the gene encoding the integral inner nuclear membrane protein emerin. Most mutations are nonsense or frameshift mutations that lead to the absence of emerin in cells. Only very few cases are due to missense or short in-frame deletions. Molecular mechanisms explaining the corresponding emerin variants' loss of function are particularly difficult to identify because of the mostly intrinsically disordered state of the emerin nucleoplasmic region. We now demonstrate that this EmN region can be produced as a disordered monomer, as revealed by nuclear magnetic resonance, but rapidly self-assembles in vitro. Increases in concentration and temperature favor the formation of long curvilinear filaments with diameters of approximately 10 nm, as observed by electron microscopy. Assembly of these filaments can be followed by fluorescence through Thioflavin-T binding and by Fourier-transform Infrared spectrometry through formation of β-structures. Analysis of the assembly properties of five EmN variants reveals that del95-99 and Q133H impact filament assembly capacities. In cells, these variants are located at the nuclear envelope, but the corresponding quantities of emerin-emerin and emerin-lamin proximities are decreased compared to wild-type protein. Furthermore, variant P183H favors EmN aggregation in vitro, and variant P183T provokes emerin accumulation in cytoplasmic foci in cells. Substitution of residue Pro183 might systematically favor oligomerization, leading to emerin aggregation and mislocalization in cells. Our results suggest that emerin self-assembly is necessary for its proper function and that a loss of either the protein itself or its ability to self-assemble causes muscular dystrophy.
Published: 2015
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2. Predictive factors of long-term outcomes of surgery for mesial temporal lobe epilepsy associated with hippocampal sclerosis

Author: Stéphane Clemenceau, Anne Bertrand, Vi-Huong Nguyen-Michel, Sophie Dupont, Franck Bielle, Ana Laura Calderón-Garcidueñas, Bertrand Mathon, Michel Baulac, Alexandre Carpentier, Vincent Navarro, Odile Plaisant, Philippe Cornu, Charles Duyckaerts, Séverine Samson, Virginie Lambrecq, Richard B. Miles, Claude Adam, Service de Neurochirurgie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute ( ICM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -CHU Pitié-Salpêtrière [APHP]-Centre National de la Recherche Scientifique ( CNRS ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ), Institut de Biologie et de Technologies de Saclay ( IBITECS ), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), CHU Pitié-Salpêtrière [APHP], Algorithms, models and methods for images and signals of the human brain ( ARAMIS ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute ( ICM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -CHU Pitié-Salpêtrière [APHP]-Centre National de la Recherche Scientifique ( CNRS ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -CHU Pitié-Salpêtrière [APHP]-Centre National de la Recherche Scientifique ( CNRS ) -Inria de Paris, Institut National de Recherche en Informatique et en Automatique ( Inria ) -Institut National de Recherche en Informatique et en Automatique ( Inria ), Service de neuroradiologie diagnostique et fonctionnelle [CHU Pitié-Salpêtrière], Université Pierre et Marie Curie - Paris 6 ( UPMC ), Institut Armand Frappier ( INRS-IAF ), Institut National de la Recherche Scientifique [Québec] ( INRS ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut Armand Frappier, CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Algorithms, models and methods for images and signals of the human brain (ARAMIS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS)-Inria de Paris, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria), Service de Neuroradiologie [CHU Pitié-Salpêtrière], Université Pierre et Marie Curie - Paris 6 (UPMC), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Inria de Paris, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Inria de Paris, and Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS)
Subjects: Adult, Male, medicine.medical_specialty, Adolescent, medicine.medical_treatment, [SDV]Life Sciences [q-bio], Neuropathology, Status epilepticus, Hippocampus, 03 medical and health sciences, Young Adult, 0302 clinical medicine, Psychiatric history, Postoperative Complications, Predictive Value of Tests, medicine, Humans, Epilepsy surgery, Longitudinal Studies, Anterior temporal lobectomy, Aged, Retrospective Studies, Hippocampal sclerosis, Sclerosis, [ SDV ] Life Sciences [q-bio], business.industry, Retrospective cohort study, Cognition, Middle Aged, medicine.disease, Anterior Temporal Lobectomy, 3. Good health, Surgery, nervous system diseases, Treatment Outcome, Neurology, Epilepsy, Temporal Lobe, 030220 oncology & carcinogenesis, Anesthesia, Female, Neurology (clinical), medicine.symptom, business, Cognition Disorders, 030217 neurology & neurosurgery
Abstract: International audience; OBJECTIVE:The reasons for failure of surgical treatment for mesial temporal lobe epilepsy (MTLE) associated with hippocampal sclerosis (HS) remain unclear. This retrospective study analyzed seizure, cognitive, and psychiatric outcomes, searching for factors associated with seizure relapse or cognitive and psychiatric deterioration after MTLE-HS surgery.METHODS:Seizure, cognitive, and psychiatric outcomes were reviewed after 389 surgeries performed between 1990 and 2015 on patients aged 15-67 years at a tertiary center. Three surgical approaches were used: anterior temporal lobectomy (ATL; n = 209), transcortical selective amygdalohippocampectomy (SAH; n = 144), and transsylvian SAH (n = 36).RESULTS:With an average follow-up of 8.7 years (range = 1.0-25.2), seizure outcome was classified as Engel I in 83.7% and Engel Ia in 57.1% of patients. The histological classification of HS was type 1 for 75.3% of patients, type 2 for 18.7%, and type 3 for 1.2%. Two factors were significantly associated with seizure recurrence: past history of status epilepticus and preoperative intracranial electroencephalographic recording. In contrast, neither HS type, the presence of a dual pathology, nor surgical approach was associated with seizure outcome. Risk of cognitive impairment was 3.12 (95% confidence interval = 1.27-7.70), greater in patients after ATL than in patients after transcortical SAH. A presurgical psychiatric history and postoperative cognitive impairment were associated with poor psychiatric outcome.SIGNIFICANCE:The SAH and ATL approaches have similar beneficial effects on seizure control, whereas transcortical SAH tends to minimize cognitive deterioration after surgery. Variation in postsurgical outcome with the class of HS should be investigated further.
Published: 2017
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3. Emerin self-assembly mechanism: role of the LEM domain

Author: Ana-Andreea Arteni, Isaline Herrada, François-Xavier Theillet, Catherine Coirault, Florian Celli, Camille Samson, Sophie Zinn-Justin, Kitty Hendriks, Adam Lange, Béatrice Alpha-Bazin, Nada Essawy, Jean Armengaud, Maximilian Zinke, Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Leibniz-Institut für Molekulare Pharmakologie, Centre de recherche en myologie, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Association française contre les myopathies ( AFM-Téléthon ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Biologie et de Technologies de Saclay ( IBITECS ), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Laboratoire de Biochimie des Systèmes Perturbés ( LBSP ), Institut für Biologie/Mikrobiologie, Humboldt Universität zu Berlin, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Leibniz Forschungsinstitut für Molekulare Pharmakolgie = Leibniz Institute for Molecular Pharmacology [Berlin, Allemagne] (FMP), Leibniz Association, Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire de Biochimie des Systèmes Perturbés (LBSP), Humboldt University Of Berlin, Coirault, Catherine, Université Pierre et Marie Curie - Paris 6 (UPMC)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Humboldt-Universität zu Berlin, and Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Université Pierre et Marie Curie - Paris 6 (UPMC)
Subjects: 0301 basic medicine, Models, Molecular, Protein Conformation, alpha-Helical, folding, Conformational change, [SDV]Life Sciences [q-bio], Mutant, Gene Expression, medicine.disease_cause, Biochemistry, lamin, Cloning, Molecular, chemistry.chemical_classification, Mutation, Nuclear Proteins, nuclear envelope, oligomerization Correspondence, intrinsically disordered region, Chromatin, Amino acid, [SDV] Life Sciences [q-bio], DNA-Binding Proteins, Protein Binding, nucleoskeleton, Recombinant Fusion Proteins, Emerin, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, oligomerization, 03 medical and health sciences, medicine, Escherichia coli, Inner membrane, Humans, Protein Interaction Domains and Motifs, Amino Acid Sequence, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Binding Sites, [ SDV ] Life Sciences [q-bio], Membrane Proteins, Cell Biology, Kinetics, 030104 developmental biology, chemistry, Biophysics, Protein Conformation, beta-Strand, Protein Multimerization, Sequence Alignment, Lamin
Abstract: International audience; At the nuclear envelope, the inner nuclear membrane protein emerin contributes to the interface between the nucleoskeleton and the chromatin. Emerin is an essential actor of the nuclear response to a mechanical signal. Genetic defects in emerin cause Emery-Dreifuss muscular dystrophy. It was proposed that emerin oligomerization regulates nucleoskeleton binding , and impaired oligomerization contributes to the loss of function of emerin disease-causing mutants. We here report the first structural characterization of emerin oligomers. We identified an N-terminal emerin region from amino acid 1 to amino acid 132 that is necessary and sufficient for formation of long curvilinear filaments. In emerin monomer, this region contains a globular LEM domain and a fragment that is intrinsically disordered. Solid-state nuclear magnetic resonance analysis identifies the LEM b-fragment as part of the oligomeric structural core. However, the LEM domain alone does not self-assemble into filaments. Additional residues forming a b-structure are observed within the filaments that could correspond to the unstructured region in emerin monomer. We show that the delK37 mutation causing muscular dystrophy triggers LEM domain unfolding and increases emerin self-assembly rate. Similarly, inserting a disulfide bridge that stabilizes the LEM folded state impairs emerin N-terminal region self-assembly, whereas reducing this disulfide bridge triggers self-assembly. We conclude that the LEM domain, responsible for binding to the chromatin protein BAF, undergoes a conformational change during self-assembly of emerin N-terminal region. The consequences of these structural rearrangement and self-assembly events on emerin binding properties are discussed. Abbreviations BAF, barrier-to-autointegration factor; DTT, dithiothreitol; EDMD, Emery-Dreifuss muscular dystrophy; LEM, Lap2-emerin-Man1; LINC, linker of nucleoskeleton and cytoskeleton; NMR, nuclear magnetic resonance; SDS/PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; ThT, thioflavin T.
Published: 2017
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4. 1H, 13C and 15N backbone resonance assignment of the intrinsically disordered region of the nuclear envelope protein emerin

Author: Florian Celli, Sophie Zinn-Justin, Isaline Herrada, Camille Samson, Francois-Xavier Theillet, Institut de Biologie et de Technologies de Saclay ( IBITECS ), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Institut de Biologie et de Technologies de Saclay (IBITECS), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay
Subjects: 0301 basic medicine, [SDV]Life Sciences [q-bio], Emerin, 010402 general chemistry, 01 natural sciences, Biochemistry, Nuclear envelope, 03 medical and health sciences, Residue (chemistry), NMR spectroscopy, Structural Biology, Inner membrane, Humans, Urea, Nuclear protein, Nuclear Magnetic Resonance, Biomolecular, Dose-Response Relationship, Drug, [ SDV ] Life Sciences [q-bio], Chemistry, Membrane Proteins, Nuclear Proteins, Nuclear magnetic resonance spectroscopy, Lamin Type A, Muscular dystrophy, 0104 chemical sciences, Chromatin, 030104 developmental biology, Intrinsically disordered protein, Biophysics, Lamin, Nuclear localization sequence
Abstract: International audience; Human emerin is an inner nuclear membrane protein involved in the response of the nucleus to mechanical stress. It contributes to the physical connection between the cytoskeleton and the nucleoskeleton. It is also involved in chromatin organization. Its N-terminal region is nucleoplasmic and comprises a globular LEM domain from residue 1 to residue 43. The three-dimensional structure of this LEM domain in complex with the chromatin BAF protein was solved from NMR data. Apart from the LEM domain, the nucleoplasmic region of emerin, from residue 44 to residue 221, is predicted to be intrinsically disordered. Mutations in this region impair binding to several emerin partners as lamin A, actin or HDAC3. However the molecular details of these recognition defects are unknown. Here we report (1)H, (15)N, (13)CO, (13)Cα and (13)Cβ NMR chemical shift assignments of the emerin fragment from residue 67 to residue 170, which is sufficient for nuclear localization and involved in lamin A binding. Chemical shift analysis confirms that this fragment is intrinsically disordered in 0 and 8 M urea.
Published: 2016
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5. Structural rearrangements in the phage head-to-tail interface during assembly and infection

Author: Paulo Tavares, Charlène Cornilleau, Sophie Zinn-Justin, Elena V. Orlova, Rudolf Lurz, S. Brasilès, Matthia A. Karreman, Yuriy Chaban, Institute of Structural & Molecular Biology, Birkbeck College, University College of London [London] ( UCL ), Max-Planck-Institut für Molekulare Genetik ( MPIMG ), Bactériophage T5 ( T5PHAG ), Département Virologie ( Dpt Viro ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ), European Molecular Biology Laboratory [Heidelberg] ( EMBL ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Lab Virol Mol & Struct, Ctr Rech Gif, UPR 3296, Centre National de la Recherche Scientifique ( CNRS ), University College of London [London] (UCL), Max-Planck-Institut für Molekulare Genetik (MPIMG), Max-Planck-Gesellschaft, Bactériophage T5 (T5PHAG), Département Virologie (Dpt Viro), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), European Molecular Biology Laboratory [Heidelberg] (EMBL), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, Protein Conformation, Viral protein, [SDV]Life Sciences [q-bio], viruses, Genome, Viral, Biology, medicine.disease_cause, Genome, Virus, Bacteriophage, Viral Proteins, 03 medical and health sciences, chemistry.chemical_compound, Protein structure, medicine, Bacteriophages, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Genetics, 0303 health sciences, Multidisciplinary, [ SDV ] Life Sciences [q-bio], Virus Assembly, Cryoelectron Microscopy, 030302 biochemistry & molecular biology, Viral Tail Proteins, Biological Sciences, Virus Internalization, biology.organism_classification, 3. Good health, Cell biology, chemistry, Capsid, DNA
Abstract: Many icosahedral viruses use a specialized portal vertex to control genome encapsidation and release from the viral capsid. In tailed bacteriophages, the portal system is connected to a tail structure that provides the pipeline for genome delivery to the host cell. We report the first, to our knowledge, subnanometer structures of the complete portal-phage tail interface that mimic the states before and after DNA release during phage infection. They uncover structural rearrangements associated with intimate protein-DNA interactions. The portal protein gp6 of bacteriophage SPP1 undergoes a concerted reorganization of the structural elements of its central channel during interaction with DNA. A network of protein-protein interactions primes consecutive binding of proteins gp15 and gp16 to extend and close the channel. This critical step that prevents genome leakage from the capsid is achieved by a previously unidentified allosteric mechanism: gp16 binding to two different regions of gp15 drives correct positioning and folding of an inner gp16 loop to interact with equivalent loops of the other gp16 subunits. Together, these loops build a plug that closes the channel. Gp16 then fastens the tail to yield the infectious virion. The gatekeeper system opens for viral genome exit at the beginning of infection but recloses afterward, suggesting a molecular diaphragm-like mechanism to control DNA efflux. The mechanisms described here, controlling the essential steps of phage genome movements during virus assembly and infection, are likely to be conserved among long-tailed phages, the largest group of viruses in the Biosphere.
Published: 2015
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6. Bacteriophage SPP1 tail tube protein self-assembles into β-structure-rich tubes

Author: Langlois, Chantal, Ramboarina, Stéphanie, Cukkemane, Abhishek, Auzat, Isabelle, Chagot, Benjamin, Gilquin, Bernard, Ignatiou, Athanasios, Petitpas, Isabelle, Kasotakis, Emmanouil, Paternostre, Maïté, White, Helen E., Orlova, Elena V., Baldus, Marc, Tavares, Paulo, Zinn-Justin, Sophie, Sub NMR Spectroscopy, NMR Spectroscopy, Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Department of Chemistry, Faculty of Science, Utrecht University, 3584 CH Utrecht, The Netherlands, Laboratoire de Virologie Moléculaire et Structurale, Centre National de la Recherche Scientifique (CNRS), Institute of Structural & Molecular Biology, Birkbeck College, University College of London [London] (UCL), Interactions et mécanismes d’assemblage des protéines et des peptides (IMAPP), Département Biochimie, Biophysique et Biologie Structurale (B3S), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie et de Technologies de Saclay ( IBITECS ), Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Centre National de la Recherche Scientifique ( CNRS ), University College of London [London] ( UCL ), Interactions et mécanismes d’assemblage des protéines et des peptides ( IMAPP ), Département Biochimie, Biophysique et Biologie Structurale ( B3S ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ), Sub NMR Spectroscopy, and NMR Spectroscopy
Subjects: Protein Folding, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Fourier Transform IR (FTIR), Biology, Biochemistry, Bacteriophage, Viral Proteins, Tube (fluid conveyance), Bacteriophages, Electron Microscopy, cardiovascular diseases, Amino Acid Sequence, Peptide sequence, Molecular Biology, [ SDV ] Life Sciences [q-bio], Tertiary Structure, Virus Assembly, Virion, Computational Biology, Nuclear magnetic resonance spectroscopy, Cell Biology, biology.organism_classification, Solid State NMR, Protein tertiary structure, In vitro, Protein Structure, Tertiary, Crystallography, Capsid, Tail Tube, cardiovascular system, Protein folding
Abstract: International audience; The majority of known bacteriophages have long tails that serve for bacterial target recognition and viral DNA delivery into the host. These structures form a tube from the viral capsid to the bacterial cell. The tube is formed primarily by a helical array of tail tube protein (TTP) subunits. In phages with a contractile tail, the TTP tube is surrounded by a sheath structure. Here, we report the first evidence that a phage TTP, gp17.1 of siphophage SPP1, self-assembles into long tubes in the absence of other viral proteins. gp17.1 does not exhibit a stable globular structure when monomeric in solution, even if it was confidently predicted to adopt the β-sandwich fold of phage λ TTP. However, Fourier transform infrared and nuclear magnetic resonance spectroscopy analyses showed that its β-sheet content increases significantly during tube assembly, suggesting that gp17.1 acquires a stable β-sandwich fold only after self-assembly. EM analyses revealed that the tube is formed by hexameric rings stacked helicoidally with the same organization and helical parameters found for the tail of SPP1 virions. These parameters were used to build a pseudo-atomic model of the TTP tube. The large loop spanning residues 40-56 is located on the inner surface of the tube, at the interface between adjacent monomers and hexamers. In line with our structural predictions, deletion of this loop hinders gp17.1 tube assembly in vitro and interferes with SPP1 tail assembly during phage particle morphogenesis in bacteria.
Published: 2015
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7. Structural characterization of filaments formed by human Xrcc4-Cernunnos/XLF complex involved in nonhomologous DNA end-joining

Author: Jean-Pierre de Villartay, Pierre Legrand, José A. Márquez, Eric Le Cam, Isabelle Callebaut, Guilhem Faure, Raphael Guerois, Olivier Piétrement, Jean-Baptiste Charbonnier, Sonia Baconnais, Pascal Drevet, Virginie Ropars, Jeremy Amram, Patrick Revy, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Hôpital Henri Mondor, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Interfaces biomatériaux/tissus hôtes, Université de Reims Champagne-Ardenne (URCA)-IFR53-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Gustave Roussy (IGR), CCADET, Universidad Nacional Autónoma de México (UNAM), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Système membranaires, photobiologie, stress et détoxication (SMPSD), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM), Institut National de la Santé et de la Recherche Médicale (INSERM), Universidad Nacional Autónoma de México = National Autonomous University of Mexico (UNAM), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Maintenance des génomes, Microscopies Moléculaire et Bionanosciences, Centre National de la Recherche Scientifique-Université Paris Sud, Maintenance des génomes, High Throughput Crystallization Laboratory, European Molecular Biology Laboratory Grenoble Outstation, European Molecular Biology Laboratory, Institut de minéralogie et de physique des milieux condensés (IMPMC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Institut de Physique du Globe de Paris (IPG Paris)-Centre National de la Recherche Scientifique (CNRS), Developpement Normal et Pathologique du Système Immunitaire, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie ( LBSR ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Synchrotron SOLEIL ( SSOLEIL ), Centre National de la Recherche Scientifique ( CNRS ), Institut de minéralogie et de physique des milieux condensés ( IMPMC ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -IPG PARIS-Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Université Pierre et Marie Curie - Paris 6 (UPMC)-IPG PARIS-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, DNA Repair, DNA repair, Phenylalanine, [SDV]Life Sciences [q-bio], Blotting, Western, Molecular Sequence Data, Glutamic Acid, Calorimetry, Biology, LIG4, Crystallography, X-Ray, medicine.disease_cause, DNA-binding protein, 03 medical and health sciences, chemistry.chemical_compound, Methionine, 0302 clinical medicine, Microscopy, Electron, Transmission, medicine, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, chemistry.chemical_classification, Aspartic Acid, 0303 health sciences, Mutation, DNA ligase, Binding Sites, Multidisciplinary, Sequence Homology, Amino Acid, Mutagenesis, Biological Sciences, DNA repair protein XRCC4, Peptide Fragments, Protein Structure, Tertiary, DNA-Binding Proteins, DNA Repair Enzymes, chemistry, Biochemistry, Biophysics, Protein Multimerization, 030217 neurology & neurosurgery, DNA, Protein Binding
Abstract: Cernunnos/XLF is a core protein of the nonhomologous DNA end-joining (NHEJ) pathway that processes the majority of DNA double-strand breaks in mammals. Cernunnos stimulates the final ligation step catalyzed by the complex between DNA ligase IV and Xrcc4 (X4). Here we present the crystal structure of the X 4 1–157 -Cernunnos 1–224 complex at 5.5-Å resolution and identify the relative positions of the two factors and their binding sites. The X-ray structure reveals a filament arrangement for X 4 1–157 and Cernunnos 1–224 homodimers mediated by repeated interactions through their N-terminal head domains. A filament arrangement of the X4–Cernunnos complex was confirmed by transmission electron microscopy analyses both with truncated and full-length proteins. We further modeled the interface and used structure-based site-directed mutagenesis and calorimetry to characterize the roles of various residues at the X4–Cernunnos interface. We identified four X4 residues (Glu 55 , Asp 58 , Met 61 , and Phe 106 ) essential for the interaction with Cernunnos. These findings provide new insights into the molecular bases for stimulatory and bridging roles of Cernunnos in the final DNA ligation step.
Published: 2011
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8. Structural and functional insights into Archaeoglobus fulgidus m2G10 tRNA methyltransferase Trm11 and its Trm112 activator

Author: Can Wang, Nhan van Tran, Marc Graille, Vincent Guérineau, Vincent Jactel, Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de synthèse organique (DCSO), École polytechnique (X)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Substances Naturelles (ICSN), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), ANR-14-CE09-0016,TrMTases,Trm112, un activateur de methyltransférases à l'interface entre la biogenèse du ribosome et sa fonction(2014), Laboratoire Stockage Optique (LSO), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: Models, Molecular, Protein Conformation, AcademicSubjects/SCI00010, Archaeal Proteins, [SDV]Life Sciences [q-bio], RNA, Archaeal, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, 03 medical and health sciences, Protein structure, RNA, Transfer, Genetics, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Gene, 030304 developmental biology, tRNA Methyltransferases, 0303 health sciences, Molecular Structure, Nucleic Acid Enzymes, Activator (genetics), 030302 biochemistry & molecular biology, Archaeoglobus fulgidus, TRNA Methyltransferase, Translation (biology), TRNA Methyltransferases, Biochemistry, Transfer RNA, Protein Processing, Post-Translational, Protein Binding
Abstract: tRNAs play a central role during the translation process and are heavily post-transcriptionally modified to ensure optimal and faithful mRNA decoding. These epitranscriptomics marks are added by largely conserved proteins and defects in the function of some of these enzymes are responsible for neurodevelopmental disorders and cancers. Here, we focus on the Trm11 enzyme, which forms N2-methylguanosine (m2G) at position 10 of several tRNAs in both archaea and eukaryotes. While eukaryotic Trm11 enzyme is only active as a complex with Trm112, an allosteric activator of methyltransferases modifying factors (RNAs and proteins) involved in mRNA translation, former studies have shown that some archaeal Trm11 proteins are active on their own. As these studies were performed on Trm11 enzymes originating from archaeal organisms lacking TRM112 gene, we have characterized Trm11 (AfTrm11) from the Archaeoglobus fulgidus archaeon, which genome encodes for a Trm112 protein (AfTrm112). We show that AfTrm11 interacts directly with AfTrm112 similarly to eukaryotic enzymes and that although AfTrm11 is active as a single protein, its enzymatic activity is strongly enhanced by AfTrm112. We finally describe the first crystal structures of the AfTrm11-Trm112 complex and of Trm11, alone or bound to the methyltransferase inhibitor sinefungin.
Published: 2020
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9. Division of labor in epitranscriptomics: What have we learnt from the structures of eukaryotic and viral multimeric RNA methyltransferases?

Author: Marc Graille, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0303 health sciences, Messenger RNA, [SDV]Life Sciences [q-bio], 030302 biochemistry & molecular biology, Eukaryota, RNA, Translation (biology), Methyltransferases, Computational biology, Biology, Biochemistry, Ribosome, 03 medical and health sciences, RNA, Transfer, Holoenzymes, RNA editing, Epitranscriptomics, Humans, RNA, Viral, RNA Processing, Post-Transcriptional, Molecular Biology, Gene, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology
Abstract: The translation of an mRNA template into the corresponding protein is a highly complex and regulated choreography performed by ribosomes, tRNAs, and translation factors. Most RNAs involved in this process are decorated by multiple chemical modifications (known as epitranscriptomic marks) contributing to the efficiency, the fidelity, and the regulation of the mRNA translation process. Many of these epitranscriptomic marks are written by holoenzymes made of a catalytic subunit associated with an activating subunit. These holoenzymes play critical roles in cell development. Indeed, several mutations being identified in the genes encoding for those proteins are linked to human pathologies such as cancers and intellectual disorders for instance. This review describes the structural and functional properties of RNA methyltransferase holoenzymes, which when mutated often result in brain development pathologies. It illustrates how structurally different activating subunits contribute to the catalytic activity of these holoenzymes through common mechanistic trends that most likely apply to other classes of holoenzymes. This article is categorized under: RNA Processing > RNA Editing and Modification RNA Processing > Capping and 5' End Modifications.
Published: 2021
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10. Exposure to the Methylselenol Precursor Dimethyldiselenide Induces a Reductive Endoplasmic Reticulum Stress in Saccharomyces cerevisiae

Author: Pierre Plateau, Marc Dauplais, Myriam Lazard, Pierre Mahou, Laboratoire de Biochimie de l'Ecole polytechnique (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Optique et Biosciences (LOB), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École polytechnique (X), Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Lazard, Myriam
Subjects: QH301-705.5, [SDV]Life Sciences [q-bio], Saccharomyces cerevisiae, Mutant, Catalysis, Inorganic Chemistry, 03 medical and health sciences, 0302 clinical medicine, Oxidoreductase, Physical and Theoretical Chemistry, Biology (General), selenium, Molecular Biology, QD1-999, Spectroscopy, 030304 developmental biology, YAP1, chemistry.chemical_classification, 0303 health sciences, biology, Cell growth, Chemistry, Endoplasmic reticulum, Oxidative folding, disulfide bond formation, Organic Chemistry, reductive stress, General Medicine, unfolded protein response, biology.organism_classification, methylselenol, Computer Science Applications, Cell biology, [SDV] Life Sciences [q-bio], dimethyldiselenide, oxidative folding, Unfolded protein response, ER stress, 030217 neurology & neurosurgery
Abstract: Methylselenol (MeSeH) is a major cytotoxic metabolite of selenium, causing apoptosis in cancer cells through mechanisms that remain to be fully established. Previously, we demonstrated that, in Saccharomyces cerevisiae, MeSeH toxicity was mediated by its metabolization into selenomethionine by O-acetylhomoserine (OAH)-sulfhydrylase, an enzyme that is absent in higher eukaryotes. In this report, we used a mutant met17 yeast strain, devoid of OAH- sulfhydrylase activity, to identify alternative targets of MeSeH. Exposure to dimethyldiselenide (DMDSe), a direct precursor of MeSeH, caused an endoplasmic reticulum (ER) stress, as evidenced by increased expression of the ER chaperone Kar2p. Mutant strains (∆ire1 and ∆hac1) unable to activate the unfolded protein response were hypersensitive to MeSeH precursors but not to selenomethionine. In contrast, deletion of YAP1 or SKN7, required to activate the oxidative stress response, did not affect cell growth in the presence of DMDSe. ER maturation of newly synthesized carboxypeptidase Y was impaired, indicating that MeSeH/DMDSe caused protein misfolding in the ER. Exposure to DMDSe resulted in induction of the expression of the ER oxidoreductase Ero1p with concomitant reduction of its regulatory disulfide bonds. These results suggest that MeSeH disturbs protein folding in the ER by generating a reductive stress in this compartment.
Published: 2021
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11. A scaffold lncRNA shapes the mitosis to meiosis switch

Author: Mathieu Rougemaille, Sylvie Auxilien, Ditipriya Hazra, Vedrana Andric, Alicia Nevers, Marc Graille, Benoit Palancade, Alexandra Menant, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), University of Oxford [Oxford], Institut Jacques Monod (IJM (UMR_7592)), Université de Paris (UP)-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE11-0003,m6A,La modification m6A des ANRm: biogenèse et rôle dans la stabilité des ARNm(2016), ANR-18-CE12-0003,CisTransRloop,Facteurs contrôlant l'instabilité génétique associée aux R-loops en cis et en trans(2018), ANR-16-CE12-0031,MechaMiMe,Mécanismes de la transition mitose-méiose chez la levure fissipare Schizosaccharomyces pombe(2016), Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Protéines de liaison à l’ARN dans l’expression des gènes et la différenciation cellulaire (GEXDIF), Département Biologie des Génomes (DBG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)
Subjects: 0301 basic medicine, Science, [SDV]Life Sciences [q-bio], RNA Stability, General Physics and Astronomy, Mitosis, Biology, RNA decay, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, 0302 clinical medicine, Downregulation and upregulation, Meiosis, Gene Expression Regulation, Fungal, Gene expression, Schizosaccharomyces, RNA, Messenger, X-ray crystallography, Regulation of gene expression, mRNA Cleavage and Polyadenylation Factors, Multidisciplinary, Effector, Fungal genetics, RNA, RNA-Binding Proteins, RNA, Fungal, General Chemistry, 3. Good health, Cell biology, 030104 developmental biology, Long non-coding RNAs, RNA, Long Noncoding, Schizosaccharomyces pombe Proteins, 030217 neurology & neurosurgery, Protein Binding
Abstract: Long non-coding RNAs (lncRNAs) contribute to the regulation of gene expression in response to intra- or extracellular signals but the underlying molecular mechanisms remain largely unexplored. Here, we identify an uncharacterized lncRNA as a central player in shaping the meiotic gene expression program in fission yeast. We report that this regulatory RNA, termed mamRNA, scaffolds the antagonistic RNA-binding proteins Mmi1 and Mei2 to ensure their reciprocal inhibition and fine tune meiotic mRNA degradation during mitotic growth. Mechanistically, mamRNA allows Mmi1 to target Mei2 for ubiquitin-mediated downregulation, and conversely enables accumulating Mei2 to impede Mmi1 activity, thereby reinforcing the mitosis to meiosis switch. These regulations also occur within a unique Mmi1-containing nuclear body, positioning mamRNA as a spatially-confined sensor of Mei2 levels. Our results thus provide a mechanistic basis for the mutual control of gametogenesis effectors and further expand our vision of the regulatory potential of lncRNAs., In fission yeast, the antagonistic RNA-binding proteins Mmi1 and Mei2 respectively promote and inhibit meiotic mRNA degradation during mitotic growth. Here the authors show that the lncRNA mamRNA scaffolds Mmi1 and Mei2 proteins to enable their mutual controls.
Published: 2021
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12. The Arp1/11 minifilament of dynactin primes the endosomal Arp2/3 complex

Author: Frédéric Saudou, Emmanuel Derivery, Ksenia Oguievetskaia, Andrew P. Carter, Maria-Victoria Hinckelmann, Nathalie Rocques, Christophe Le Clainche, Guillaume Romet-Lemonne, Véronique Henriot, Nicolas Molinie, Artem I. Fokin, Alexis Gautreau, Violaine David, Luyan Cao, Magali Aumont-Nicaise, Caroline E. Stone, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Mesures d'interactions protéine-protéine (PIM), Département Plateforme (PF I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Dynamique du cytosquelette et motilité (ACTIN), Département Biochimie, Biophysique et Biologie Structurale (B3S), Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), MRC Laboratory of Molecular Biology [Cambridge, UK] (LMB), University of Cambridge [UK] (CAM)-Medical Research Council, Institut Jacques Monod (IJM (UMR_7592)), Université de Paris (UP)-Centre National de la Recherche Scientifique (CNRS), [GIN] Grenoble Institut des Neurosciences (GIN), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA), Moscow Institute of Physics and Technology [Moscow] (MIPT), Le Clainche, Christophe, and Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)
Subjects: Endosome, Protein subunit, [SDV]Life Sciences [q-bio], Mutant, Arp2/3 complex, macromolecular substances, Biochemistry, WASH complex, Protein filament, 03 medical and health sciences, 0302 clinical medicine, Research Articles, Actin, 030304 developmental biology, 0303 health sciences, Multidisciplinary, biology, urogenital system, Chemistry, fungi, SciAdv r-articles, 3. Good health, [SDV] Life Sciences [q-bio], Dynactin, biology.protein, Biophysics, 030217 neurology & neurosurgery, Research Article
Abstract: Dendritic actin networks grow in an autocatalytic manner starting from the uncapped minifilament of dynactin., Dendritic actin networks develop from a first actin filament through branching by the Arp2/3 complex. At the surface of endosomes, the WASH complex activates the Arp2/3 complex and interacts with the capping protein for unclear reasons. Here, we show that the WASH complex interacts with dynactin and uncaps it through its FAM21 subunit. In vitro, the uncapped Arp1/11 minifilament elongates an actin filament, which then primes the WASH-induced Arp2/3 branching reaction. In dynactin-depleted cells or in cells where the WASH complex is reconstituted with a FAM21 mutant that cannot uncap dynactin, formation of branched actin at the endosomal surface is impaired. Our results reveal the importance of the WASH complex in coordinating two complexes containing actin-related proteins.
Published: 2021
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13. Selenium metabolism and toxicity in the yeast Saccharomyces cerevisiae

Author: Myriam Lazard, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Selenocysteine, biology, [SDV]Life Sciences [q-bio], Saccharomyces cerevisiae, chemistry.chemical_element, Metabolism, biology.organism_classification, Yeast, chemistry.chemical_compound, chemistry, Biochemistry, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Toxicity, Cancer cell, Selenium metabolism, Selenium
Abstract: International audience; Selenium (Se) is an essential trace element of considerable interest in humans from both a nutritional and a toxicological perspective because of the narrow margin between intakes that result in efficacy and toxicity. It is used as selenocysteine in a few selenoproteins with important physiological functions. Moreover, at supranutritional doses, Se-containing compounds have attracted interest as potential anticancer agents with high efficacy and selectivity against cancer cells. Thus, Se is becoming a widely used dietary supplement. However, accumulating evidence indicate that adverse health effects are associated with excess dietary supplementation. Therefore, characterizing the toxicity of Se metabolic intermediates are important steps to better understand both the beneficial and toxic mechanisms of Se. This review focuses on the metabolism of Se and the biological mechanisms explaining the toxicity of important Se-metabolites in the yeast Saccharomyces cerevisiae, which can be used as a model system to understand the mode of action and the biological effects of supranutritional Se in higher eukaryotes.
Published: 2021
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14. CYFIP2 containing WAVE complexes inhibit cell migration

Author: Polesskaya, Anna, Boutillon, Arthur, Wang, Yanan, Lavielle, Marc, Vacher, Sophie, Schnitzler, Anne, Molinie, Nicolas, Rocques, Nathalie, Fokin, Artem, Bièche, Ivan, David, Nicolas, Gautreau, Alexis, Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Modélisation en pharmacologie de population (XPOP), Centre de Mathématiques Appliquées - Ecole Polytechnique (CMAP), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)-École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)-Inria Saclay - Ile de France, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria), and Institut Curie [Paris]
Subjects: knock-out, Arp2/3 MFS, knock-down, Scar/WAVE, [SDV]Life Sciences [q-bio], metastasis-free survival, KD, multiprotein complexes, KO, Patient cohort, Zebrafish embryo, morpholino, MO
Abstract: Branched actin networks polymerized by the Arp2/3 complex are critical for cell migration. The WAVE complex is the major Arp2/3 activator at the leading edge of migrating cells. However, multiple distinct WAVE complexes can be assembled in a cell, due to the combinatorial complexity of paralogous subunits. When systematically analyzing the contribution of each WAVE complex subunit to the metastasis-free survival of breast cancer patients, we found that overexpression of the CYFIP2 subunit was surprisingly associated with good prognosis. Gain and loss of function experiments in transformed and untransformed mammary epithelial cells revealed that cell migration was always inversely related to CYFIP2 levels. The role of CYFIP2 was systematically opposite to the role of the paralogous subunit CYFIP1 or of the NCKAP1 subunit. The specific CYFIP2 function in inhibiting cell migration was related to its unique ability to down-regulate classical pro-migratory WAVE complexes. The anti-migratory function of CYFIP2 was also revealed in migration of prechordal plate cells during gastrulation of the zebrafish embryo, indicating that the unique function of CYFIP2 is critically important in both physiological and pathophysiological migrations.
Published: 2020

15. Vécu au travail des sages-femmes, une analyse textuelle

Author: Robin, Noluenn, Josso, Nolwenn, Launay, Titouan, Robin Paulard, Isabelle, Pougnet, Laurence, Pougnet, Richard, Laboratoire de Biologie structurale, Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Études et de Recherche en Sociologie (LABERS), Université de Bretagne Sud (UBS)-Université de Brest (UBO)-Institut Brestois des Sciences de l'Homme et de la Société (IBSHS), Université de Brest (UBO), and Hôpital Morvan - CHRU de Brest (CHU - BREST )
Subjects: [SDV]Life Sciences [q-bio]
Published: 2020
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16. microRNA-induced translational control of antiviral immunity by the cap-binding protein 4EHP

Author: Rongtuan Lin, Angela P. Hackett, Sung-Hoon Kim, David L. Dai, Tommy Alain, Seyed Mehdi Jafarnejad, Isabela Fabri Karam, Nahum Sonenberg, Long Yang, Clément Chapat, Jung-Hyun Choi, Shane Wiebe, Jun Luo, Nathaniel Robichaud, Qian Li, Peng Wang, Xu Zhang, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], Mice, Transgenic, Biology, 03 medical and health sciences, Mice, 0302 clinical medicine, RNA Virus Infections, Downregulation and upregulation, microRNA, Gene silencing, Animals, Humans, RNA Viruses, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Messenger RNA, Innate immune system, Binding protein, Translation (biology), Cell Biology, Interferon-beta, Immunity, Innate, 3. Good health, Cell biology, MicroRNAs, Eukaryotic Initiation Factor-4E, HEK293 Cells, Viral replication, Protein Biosynthesis, 030217 neurology & neurosurgery
Abstract: Summary Type I interferons (IFNs) are critical cytokines in the host defense against invading pathogens. Sustained production of IFNs, however, is detrimental to the host, as it provokes autoimmune diseases. Thus, the expression of IFNs is tightly controlled. We report that the mRNA 5′ cap-binding protein 4EHP plays a key role in regulating type I IFN concomitant with controlling virus replication, both in vitro and in vivo. Mechanistically, 4EHP suppresses IFN-β production by effecting the miR-34a-induced translational silencing of Ifnb1 mRNA. miR-34a is upregulated by both RNA virus infection and IFN-β induction, prompting a negative feedback regulatory mechanism that represses IFN-β expression via 4EHP. These findings demonstrate the direct involvement of 4EHP in virus-induced host response, underscoring a critical translational silencing mechanism mediated by 4EHP and miR-34a to impede sustained IFN production. This study highlights an intrinsic regulatory function for miRNA and the translation machinery in maintaining host homeostasis.
Published: 2020
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17. The Long and Winding Road Toward Efficient High-Performance Computing

Author: William Jalby, Michel Masella, David Kuck, Allen D. Malony, Mihail Popov, Abdelhafid Mazouz, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Laboratoire d'Informatique Parallélisme Réseaux Algorithmes Distribués (LI-PaRAD), Computer Science Department [Oregon], University of Oregon [Eugene], Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Bull atos technologies
Subjects: Computer science, [SDV]Life Sciences [q-bio], design, molecular-dynamics, performance evaluation tools, 02 engineering and technology, INTGEN, Software, Autotuning, 0202 electrical engineering, electronic engineering, information engineering, Code (cryptography), benchmarking, Electrical and Electronic Engineering, implementation, business.industry, Benchmarking, Supercomputer, molecular dynamics, 020202 computer hardware & architecture, hardware design, Computer architecture, code, Key (cryptography), Software design, 020201 artificial intelligence & image processing, Analysis tools, business, B3S
Abstract: WOS:000448616200009; The major challenge to Exaflop computing, and more generally, efficient high-end computing, is in finding the best "matches" between advanced hardware capabilities and the software used to program applications, so that top performance will be achieved. Several benchmarks show very disappointing performance progress over the last decade, clearly indicating a mismatch between hardware and software. To remedy this problem, it is important that key performance enablers at the software level-autotuning, performance analysis tools, full application optimization-are understood. For each area, we highlight major limitations and most promising approaches to reaching better performance and energy levels. Finally, we conclude by analyzing hardware and software design, trying to pave the way for more tightly integrated hardware and software codesign.
Published: 2018
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18. Conception sur une base rationnelle de peptides de haute affinité inhibant l'histone chaperon ASF1

Author: Bakail, May, Gaubert, Albane, Andreani, Jessica, Moal, Gwenaëlle, Pinna, Guillaume, Boyarchuk, Ekaterina, Gaillard, Marie-Cécile, Courbeyrette, Régis, Mann, Carl, Thuret, Jean-Yves, Guichard, Berengère, Murciano, Brice, Richet, Nicolas, Poitou, Adeline, Frederic, Claire, Le Du, Marie-Hélène, Agez, Morgane, Roelants, Caroline, Gurard-Levin, Zachary, Almouzni, Geneviève, Cherradi, Nadia, Guérois, Raphaël, Ochsenbein, Françoise, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Assemblage moléculaire et intégrité du génome (AMIG), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), PARi (PARI), Département Plateforme (PF I2BC), Sorbonne Université - Faculté de Médecine (SU FM), Sorbonne Université (SU), Sénescence et stabilité génomique (SEN), Département Biologie des Génomes (DBG), Vaccination Antiparasitaire : Laboratoire de Biologie Cellulaire et Moléculaire (LBCM), Université Montpellier 1 (UM1)-Université de Montpellier (UM), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Invasion mechanisms in angiogenesis and cancer (IMAC), Biologie du Cancer et de l'Infection (BCI ), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM), Dynamique du noyau [Institut Curie], and Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Rosetta Design, Drug Design, [SDV]Life Sciences [q-bio], X-Ray Crystallography, Peptide Inhibitor, Epigenetics, MESH: Amino Acid Sequence, Binding Sites, Histones / metabolism, Neoplasms / drug therapy, Transplantation, Homologous, Cell Penetrating Peptide, Chromatin, Protein-Protein Interaction, Protein Binding, Cancer, [SHS]Humanities and Social Sciences
Abstract: International audience; Anti-silencing function 1 (ASF1) is a conserved H3-H4 histone chaperone involved in histone dynamics during replication, transcription, and DNA repair. Overexpressed in proliferating tissues including many tumors, ASF1 has emerged as a promising therapeutic target. Here, we combine structural, computational, and biochemical approaches to design peptides that inhibit the ASF1-histone interaction. Starting from the structure of the human ASF1-histone complex, we developed a rational design strategy combining epitope tethering and optimization of interface contacts to identify a potent peptide inhibitor with a dissociation constant of 3 nM. When introduced into cultured cells, the inhibitors impair cell proliferation, perturb cell-cycle progression, and reduce cell migration and invasion in a manner commensurate with their affinity for ASF1. Finally, we find that direct injection of the most potent ASF1 peptide inhibitor in mouse allografts reduces tumor growth. Our results open new avenues to use ASF1 inhibitors as promising leads for cancer therapy.; La fonction anti-silencing 1 (ASF1) est un chaperon d'histone H3-H4 conservé, impliqué dans la dynamique des histones pendant la réplication, la transcription et la réparation de l'ADN. Surexprimée dans les tissus en prolifération, y compris dans de nombreuses tumeurs, l'ASF1 est devenue une cible thérapeutique prometteuse. Ici, nous combinons des approches structurelles, informatiques et biochimiques pour concevoir des peptides qui inhibent l'interaction ASF1-histone. En partant de la structure du complexe ASF1-histone humain, nous avons mis au point une stratégie de conception rationnelle combinant la fixation des épitopes et l'optimisation des contacts d'interface pour identifier un puissant inhibiteur peptidique avec une constante de dissociation de 3 nM. Lorsqu'ils sont introduits dans des cellules en culture, les inhibiteurs entravent la prolifération cellulaire, perturbent la progression du cycle cellulaire et réduisent la migration et l'invasion des cellules d'une manière proportionnelle à leur affinité pour l'ASF1. Enfin, nous constatons que l'injection directe du plus puissant inhibiteur du peptide ASF1 dans les allogreffes de souris réduit la croissance des tumeurs. Nos résultats ouvrent de nouvelles voies pour utiliser les inhibiteurs de l'ASF1 comme des pistes prometteuses pour le traitement du cancer.
Published: 2019
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19. Formation of S. pombe Erh1 homodimer mediates gametogenic gene silencing and meiosis progression

Author: Ditipriya Hazra, Vedrana Andric, Marc Graille, Mathieu Rougemaille, Benoit Palancade, Laboratoire de Biochimie de l'Ecole polytechnique (BIOC), Centre National de la Recherche Scientifique (CNRS)-École polytechnique (X), Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Protéines de liaison à l’ARN dans l’expression des gènes et la différenciation cellulaire (GEXDIF), Département Biologie des Génomes (DBG), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale de la Cellule (BIOC), ANR-16-CE11-0003,m6A,La modification m6A des ANRm: biogenèse et rôle dans la stabilité des ARNm(2016), and ANR-16-CE12-0031,MechaMiMe,Mécanismes de la transition mitose-méiose chez la levure fissipare Schizosaccharomyces pombe(2016)
Subjects: Protein Conformation, [SDV]Life Sciences [q-bio], Dimer, Regulator, lcsh:Medicine, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Crystallography, X-Ray, Biochemistry, Article, chemistry.chemical_compound, 03 medical and health sciences, 0302 clinical medicine, Protein structure, Meiosis, Schizosaccharomyces, Gene silencing, Gene Silencing, lcsh:Science, Gene, Mitosis, 030304 developmental biology, RNA metabolism, mRNA Cleavage and Polyadenylation Factors, 0303 health sciences, Multidisciplinary, lcsh:R, RNA-Binding Proteins, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell cycle, Phenotype, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Yeast, Cell biology, chemistry, Multiprotein Complexes, RNA, lcsh:Q, Schizosaccharomyces pombe Proteins, Protein Multimerization, Structural biology, Carrier Proteins, 030217 neurology & neurosurgery
Abstract: Timely and accurate expression of the genetic information relies on the integration of environmental cues and the activation of regulatory networks involving transcriptional and post-transcriptional mechanisms. In fission yeast, meiosis-specific transcripts are selectively targeted for degradation during mitosis by the EMC complex, composed of Erh1, the ortholog of human ERH, and the YTH family RNA-binding protein Mmi1. Here, we present the crystal structure of Erh1 and show that it assembles as a homodimer. Mutations of amino acid residues to disrupt Erh1 homodimer formation result in loss-of-function phenotypes, similar to erh1∆ cells: expression of meiotic genes is derepressed in mitotic cells and meiosis progression is severely compromised. Interestingly, formation of Erh1 homodimer is dispensable for interaction with Mmi1, suggesting that only fully assembled EMC complexes consisting of two Mmi1 molecules bridged by an Erh1 dimer are functionally competent. We also show that Erh1 does not contribute to Mmi1-dependent down-regulation of the meiosis regulator Mei2, supporting the notion that Mmi1 performs additional functions beyond EMC. Overall, our results provide a structural basis for the assembly of the EMC complex and highlight its biological relevance in gametogenic gene silencing and meiosis progression.
Published: 2019
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20. Actin polymerization downstream of integrins: signaling pathways and mechanotransduction

Author: Christophe Le Clainche, Alexis Gautreau, Stéphane Romero, Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Dynamique du cytosquelette et motilité (ACTIN), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Moscow Institute of Physics and Technology [Moscow] (MIPT), ANR-16-CE13-0007,PHAGOMECANO,Mécanotransduction associée à la phagocytose dépendante des intégrines(2016), and ANR-18-CE13-0026,RECAMECA,Etude des liens moléculaires entre la force du cytosquelette d'actomyosine, la régulation des intégrines et les propriétés de la matrice extracellulaire grâce à une reconstitution in vitro(2018)
Subjects: Integrins, [SDV]Life Sciences [q-bio], Integrin, Formins, macromolecular substances, Biochemistry, Mechanotransduction, Cellular, Actin-Related Protein 2-3 Complex, Polymerization, Extracellular matrix, 03 medical and health sciences, Mice, 0302 clinical medicine, Cell Adhesion, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Mechanotransduction, Cell adhesion, Molecular Biology, Actin, 030304 developmental biology, 0303 health sciences, Durotaxis, biology, Chemistry, Cell Biology, VASP, Actin cytoskeleton, Actins, Cell biology, Extracellular Matrix, DNA-Binding Proteins, biology.protein, Arp2/3, actin, 030217 neurology & neurosurgery
Abstract: International audience; A cell constantly adapts to its environment. Cell decisions to survive, to proliferate or to migrate are dictated not only by soluble growth factors, but also through the direct interaction of the cell with the surrounding extracellular matrix (ECM). Integrins and their connections to the actin cytoskeleton are crucial for monitoring cell attachment and the physical properties of the substratum. Cell adhesion dynamics are modulated in complex ways by the polymerization of branched and linear actin arrays, which in turn reinforce ECM-cytoskeleton connection. This review describes the major actin regulators, Ena/VASP proteins, formins and Arp2/3 complexes, in the context of signaling pathways downstream of integrins. We focus on the specific signaling pathways that transduce the rigidity of the substrate and which control durotaxis, i.e. directed migration of cells towards increased ECM rigidity. By doing so, we highlight several recent findings on mechanotransduction and put them into a broad integrative perspective that is the result of decades of intense research on the actin cytoskeleton and its regulation.
Published: 2019
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21. Conception sur une base rationnelle de peptides de haute affinité inhibant l'histone chaperon ASF1

Author: Carl Mann, Raphael Guerois, May Bakail, Ekaterina Boyarchuk, Albane Gaubert, Marie-Cécile Gaillard, Zachary A. Gurard-Levin, Françoise Ochsenbein, Brice Murciano, Claire Frederic, Gwenaëlle Moal, Geneviève Almouzni, Jean-Yves Thuret, Régis Courbeyrette, Morgane Agez, Marie-Hélène Le Du, Nicolas Richet, Jessica Andreani, Adeline Poitou, Nadia Cherradi, Berengère Guichard, Guillaume Pinna, Caroline Roelants, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Assemblage moléculaire et intégrité du génome (AMIG), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), PARi (PARI), Département Plateforme (PF I2BC), Sénescence et stabilité génomique (SEN), Département Biologie des Génomes (DBG), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Sorbonne Université - Faculté de Médecine (SU FM), Sorbonne Université (SU), Vaccination Antiparasitaire : Laboratoire de Biologie Cellulaire et Moléculaire (LBCM), Université Montpellier 1 (UM1)-Université de Montpellier (UM), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Invasion mechanisms in angiogenesis and cancer (IMAC), Biologie du Cancer et de l'Infection (BCI ), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM), Dynamique du noyau [Institut Curie], Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: [SDV]Life Sciences [q-bio], Clinical Biochemistry, Cell Cycle Proteins, 01 natural sciences, Biochemistry, [SHS]Humanities and Social Sciences, Histones, Epitopes, Mice, Cell Movement, Neoplasms, Drug Discovery, Cancer, Mice, Inbred BALB C, biology, Chromatin, Cell biology, Histone, Molecular Medicine, Thermodynamics, Peptide Inhibitor, Female, Epigenetics, Protein Binding, DNA repair, X-Ray Crystallography, Cell Penetrating Peptide, Protein–protein interaction, Rosetta Design, Cell Line, Tumor, Animals, Humans, Transplantation, Homologous, Amino Acid Sequence, Molecular Biology, Cell Proliferation, Pharmacology, Binding Sites, 010405 organic chemistry, Cell growth, Rational design, Cell Cycle Checkpoints, MESH: Amino Acid Sequence, Drug Design, Histones / metabolism, Neoplasms / drug therapy, 0104 chemical sciences, Protein-Protein Interaction, Kinetics, biology.protein, Cell-penetrating peptide, Peptides, Molecular Chaperones
Abstract: International audience; Anti-silencing function 1 (ASF1) is a conserved H3-H4 histone chaperone involved in histone dynamics during replication, transcription, and DNA repair. Overexpressed in proliferating tissues including many tumors, ASF1 has emerged as a promising therapeutic target. Here, we combine structural, computational, and biochemical approaches to design peptides that inhibit the ASF1-histone interaction. Starting from the structure of the human ASF1-histone complex, we developed a rational design strategy combining epitope tethering and optimization of interface contacts to identify a potent peptide inhibitor with a dissociation constant of 3 nM. When introduced into cultured cells, the inhibitors impair cell proliferation, perturb cell-cycle progression, and reduce cell migration and invasion in a manner commensurate with their affinity for ASF1. Finally, we find that direct injection of the most potent ASF1 peptide inhibitor in mouse allografts reduces tumor growth. Our results open new avenues to use ASF1 inhibitors as promising leads for cancer therapy.; La fonction anti-silencing 1 (ASF1) est un chaperon d'histone H3-H4 conservé, impliqué dans la dynamique des histones pendant la réplication, la transcription et la réparation de l'ADN. Surexprimée dans les tissus en prolifération, y compris dans de nombreuses tumeurs, l'ASF1 est devenue une cible thérapeutique prometteuse. Ici, nous combinons des approches structurelles, informatiques et biochimiques pour concevoir des peptides qui inhibent l'interaction ASF1-histone. En partant de la structure du complexe ASF1-histone humain, nous avons mis au point une stratégie de conception rationnelle combinant la fixation des épitopes et l'optimisation des contacts d'interface pour identifier un puissant inhibiteur peptidique avec une constante de dissociation de 3 nM. Lorsqu'ils sont introduits dans des cellules en culture, les inhibiteurs entravent la prolifération cellulaire, perturbent la progression du cycle cellulaire et réduisent la migration et l'invasion des cellules d'une manière proportionnelle à leur affinité pour l'ASF1. Enfin, nous constatons que l'injection directe du plus puissant inhibiteur du peptide ASF1 dans les allogreffes de souris réduit la croissance des tumeurs. Nos résultats ouvrent de nouvelles voies pour utiliser les inhibiteurs de l'ASF1 comme des pistes prometteuses pour le traitement du cancer.
Published: 2019
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22. Arpin Regulates Migration Persistence by Interacting with Both Tankyrases and the Arp2/3 Complex

Author: Artem I. Fokin, Jacqueline Cherfils, Alexis Gautreau, Gleb Simanov, Valérie Campanacci, Ksenia Oguievetskaia, Irene Dang, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: cell migration, [SDV]Life Sciences [q-bio], Arpin Regulates Migration cell migration, Arp2/3 complex, Tankyrase-1, medicine.disease_cause, 0302 clinical medicine, Cell Movement, migration persistence, Tankyrases, Biology (General), ComputingMilieux_MISCELLANEOUS, Spectroscopy, 0303 health sciences, Mutation, biology, Chemistry, Cell migration, General Medicine, Computer Science Applications, Cell biology, 030220 oncology & carcinogenesis, Protein Binding, QH301-705.5, Immunoprecipitation, Two-hybrid screening, macromolecular substances, Arpin, Article, Catalysis, Inorganic Chemistry, 03 medical and health sciences, Two-Hybrid System Techniques, medicine, Humans, Binding site, Tankyrase, Physical and Theoretical Chemistry, QD1-999, Molecular Biology, Actin, 030304 developmental biology, Binding Sites, Organic Chemistry, HEK293 Cells, biology.protein, Ankyrin repeat, Carrier Proteins, Arp2/3, HeLa Cells
Abstract: During cell migration, protrusion of the leading edge is driven by the polymerization of Arp2/3-dependent branched actin networks. Migration persistence is negatively regulated by the Arp2/3 inhibitory protein Arpin. To better understand Arpin regulation in the cell, we looked for interacting partners and identified both Tankyrase 1 and 2 (TNKS) using a yeast two hybrid screen and co-immunoprecipitation with full-length Arpin as a bait. Arpin interacts with ankyrin repeats of TNKS through a C-terminal binding site on its acidic tail overlapping with the Arp2/3 binding site. To uncouple the interactions of Arpin with TNKS and Arp2/3, we introduced point mutations in the Arpin tail and attempted to rescue the increased persistence of the Arpin knock-out using random plasmid integration or compensating knock-in at the ARPIN locus. Arpin mutations impairing either Arp2/3- or TNKS-interaction were insufficient to fully abolish Arpin activity. Only the mutation that affects both interactions rendered Arpin completely inactive, suggesting the existence of two independent pathways, by which Arpin controls migration persistence. Arpin was found to dissolve liquid-liquid phase separation of TNKS upon overexpression. Together these data suggest that TNKS might be mediating the function of Arpin rather than regulating Arpin.
Published: 2021
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23. Structure of the active form of Dcp1–Dcp2 decapping enzyme bound to m7GDP and its Edc3 activator

Author: Bertrand Séraphin, Clément Charenton, Régis Back, Valerio Taverniti, Claudine Gaudon-Plesse, Marc Graille, Laboratoire de Biologie Structurale de la Cellule (BIOC), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), and Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Kluyveromyces lactis, Exonuclease, Messenger RNA, Five-prime cap, Cap binding complex, biology, Chemistry, [SDV]Life Sciences [q-bio], Protein subunit, RNA-binding protein, biology.organism_classification, Ribosome, 03 medical and health sciences, Crystallography, 030104 developmental biology, Structural Biology, Biophysics, biology.protein, Molecular Biology, ComputingMilieux_MISCELLANEOUS
Abstract: The crystal structure of the Kluyveromyces lactis decapping enzyme complex Dcp1–Dcp2 is solved in the presence of activator Edc3 and reaction product m7GDP, revealing an active conformation of the enzyme. Elimination of the 5′ cap of eukaryotic mRNAs, known as decapping, is considered to be a crucial, irreversible and highly regulated step required for the rapid degradation of mRNA by Xrn1, the major cytoplasmic 5′-3′ exonuclease. Decapping is accomplished by the recruitment of a protein complex formed by the Dcp2 catalytic subunit and its Dcp1 cofactor. However, this complex has a low intrinsic enzymatic activity and requires several accessory proteins such as the Lsm1–7 complex, Pat1, Edc1–Edc2 and/or Edc3 to be fully active. Here we present the crystal structure of the active form of the yeast Kluyveromyces lactis Dcp1–Dcp2 enzyme bound to its product (m7GDP) and its potent activator Edc3. This structure of the Dcp1–Dcp2 complex bound to a cap analog further explains previously published data on substrate binding and provides hints as to the mechanism of Edc3-mediated Dcp2 activation.
Published: 2016
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24. H-1, C-13 and N-15 backbone resonance assignment of the lamin C-terminal region specific to prelamin A

Author: Ambre Petitalot, Sophie Zinn-Justin, François-Xavier Theillet, Florian Celli, Camille Samson, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Unité de génétique et biologie des cancers (U830), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris Descartes - Paris 5 (UPD5)-Institut Curie-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)
Subjects: 0301 basic medicine, Premature aging, congenital, hereditary, and neonatal diseases and abnormalities, [SDV]Life Sciences [q-bio], Mutant, Biochemistry, Nuclear envelope, LMNA, 03 medical and health sciences, NMR spectroscopy, Structural Biology, medicine, chemistry.chemical_classification, Progeria, integumentary system, Chemistry, Point mutation, nutritional and metabolic diseases, Nucleoskeleton, Progerin, medicine.disease, mutations, Cell biology, Amino acid, 030104 developmental biology, Intrinsically disordered protein, networks, embryonic structures, Lamin
Abstract: International audience; Lamins are the main components of the nucleoskeleton. They form a protein meshwork that underlies the inner nuclear membrane. Mutations in the LMNA gene coding for A-type lamins (lamins A and C) cause a large panel of human diseases, referred to as laminopathies. These diseases include muscular dystrophies, lipodystrophies and premature aging diseases. Lamin A exhibits a C-terminal region that is different from lamin C and is post-translationally modified. It is produced as prelamin A and it is then farnesylated, cleaved, carboxymethylated and cleaved again in order to become mature lamin A. In patients with the severe Hutchinson-Gilford progeria syndrome, a specific single point mutation in LMNA leads to an aberrant splicing of the LMNA gene preventing the post-translational processing of prelamin A. This leads to the accumulation of a permanently farnesylated lamin A mutant lacking 50 amino acids named progerin. We here report the NMR H-1, N-15, (CO)-C-13, C-13 and C-13 chemical shift assignment of the C-terminal region that is specific to prelamin A, from amino acid 567 to amino acid 664. We also report the NMR H-1, N-15, (CO)-C-13, C-13 and C-13 chemical shift assignment of the C-terminal region of the progerin variant, from amino acid 567 to amino acid 614. Analysis of these chemical shift data confirms that both prelamin A and progerin C-terminal domains are largely disordered and identifies a common partially populated -helix from amino acid 576 to amino acid 585. This helix is well conserved from fishes to mammals.
Published: 2018
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25. Correction: The Translocation Domain of Botulinum Neurotoxin A Moderates the Propensity of the Catalytic Domain to Interact with Membranes at Acidic pH

Author: Anne Araye, Amélie Goudet, Julien Barbier, Sylvain Pichard, Bruno Baron, Patrick England, Javier Pérez, Sophie Zinn-Justin, Alexandre Chenal, Daniel Gillet, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biophysique Moléculaire (Plate-forme), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0303 health sciences, 03 medical and health sciences, Multidisciplinary, [SDV]Life Sciences [q-bio], lcsh:R, 030302 biochemistry & molecular biology, lcsh:Medicine, lcsh:Q, lcsh:Science, 030304 developmental biology
Abstract: International audience; [This corrects the article DOI: 10.1371/journal.pone.0153401.].
Published: 2016
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26. Structural Model of Ligand-G Protein-coupled Receptor (GPCR) Complex Based on Experimental Double Mutant Cycle Data

Author: Denis Servent, Elodie Marcon, Gilles Mourier, André Ménez, Guillaume Letellier, Bernard Gilquin, Carole Fruchart-Gaillard, Sophie Zinn-Justin, Catherine Marquer, Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service de Bioénergétique, Biologie Stucturale, et Mécanismes (SB2SM), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Muséum national d'Histoire naturelle (MNHN), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Institut du Cerveau = Paris Brain Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Allosteric modulator, Stereochemistry, [SDV]Life Sciences [q-bio], Allosteric regulation, Cell Biology, Ligand Binding Protein, Protomer, Biology, Biochemistry, Transmembrane domain, Docking (molecular), Receptor, Molecular Biology, G protein-coupled receptor
Abstract: International audience; The snake toxin MT7 is a potent and specific allosteric modulator of the human M1 muscarinic receptor (hM1). We previously characterized by mutagenesis experiments the functional determinants of the MT7-hM1 receptor interaction (FruchartGaillard, C., Mourier, G., Marquer, C., Stura, E., Birdsall, N. J.,and Servent, D. (2008) Mol. Pharmacol. 74, 1554–1563) and more recently collected evidence indicating that MT7 may bind to a dimeric form of hM1 (Marquer, C., Fruchart-Gaillard, C., Mourier, G., Grandjean, O., Girard, E., le Maire, M., Brown, S., and Servent, D. (2010) Biol. Cell 102, 409– 420). To structurally characterize the MT7-hM1 complex, we adopted a strategy combining double mutant cycle experiments and molecular modeling calculations. First, thirty-three ligand-receptor proximities were identified from the analysis of sixty-one double mutant binding affinities. Several toxin residues that are more than 25 A˚ apart still contact the same residues on the receptor. As a consequence, attempts to satisfy all the restraints by docking the toxin onto a single receptor failed. The toxin was then positioned onto two receptors during five independent flexible docking simulations. The different possible ligand and receptor extracellular loop conformations were described by performing simulations in explicit solvent. All the docking calculations converged to the same conformation of the MT7-hM1 dimer complex, satisfying the experimental restraints and in which (i) the toxin interacts with the extracellular side of the receptor, (ii) the tips of MT7 loops II and III contact one hM1 protomer, whereas the tip of loop I binds to the other protomer, and (iii) the hM1dimeric interface involves the transmembrane helices TM6 and TM7. These results structurally support the high affinity and selectivity of the MT7-hM1 interaction and highlight the atypical mode of interaction of this allosteric ligand on its G proteincoupled receptor target.
Published: 2011
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27. Stable interactions between DNA polymerase δ catalytic and structural subunits are essential for efficient DNA repair

Author: Laurent Maloisel, Serge Gangloff, Jean-Baptiste Charbonnier, Clémentine Brocas, Claudine Dherin, Institut de Radiobiologie Cellulaire et Moléculaire (IRCM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire d'Etudes de la Réparation de l'ADN (LERA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Radiobiologie moléculaire et cellulaire (RMC), Système membranaires, photobiologie, stress et détoxication (SMPSD), and Laboratoire de Radiobiologie de l'ADN (LRD)
Subjects: DNA Repair, DNA polymerase, MESH: DNA Polymerase III, [SDV]Life Sciences [q-bio], MESH: Protein Structure, Secondary, MESH: Catalytic Domain, MESH: DNA Replication, DNA-Directed DNA Polymerase, MESH: Amino Acid Sequence, Biochemistry, DNA polymerase delta, Protein Structure, Secondary, MESH: Saccharomyces cerevisiae Proteins, 0302 clinical medicine, Catalytic Domain, MESH: DNA-Directed DNA Polymerase, Pol3, Recombination, Genetic, MESH: DNA Repair, Genetics, 0303 health sciences, DNA clamp, Molecular Structure, biology, MESH: Saccharomyces cerevisiae, Cell biology, MESH: Mutagenesis, Site-Directed, MESH: Recombination, Genetic, DNA polymerase mu, Protein Binding, DNA Replication, Saccharomyces cerevisiae Proteins, DNA polymerase II, Molecular Sequence Data, MESH: Molecular Structure, Saccharomyces cerevisiae, 03 medical and health sciences, Proliferating Cell Nuclear Antigen, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Molecular Biology, Replication protein A, Alleles, DNA Polymerase III, 030304 developmental biology, MESH: Molecular Sequence Data, MESH: Alleles, DNA replication, Pol32, Cell Biology, Processivity, Pol31, MESH: Proliferating Cell Nuclear Antigen, Mutagenesis, Site-Directed, biology.protein, Genes, Lethal, MESH: Genes, Lethal, 030217 neurology & neurosurgery
Abstract: International audience; Eukaryotic DNA polymerase δ (Pol δ) activity is crucial for chromosome replication and DNA repair and thus, plays an essential role in genome stability. In Saccharomyces cerevisiae, Pol δ is a heterotrimeric complex composed of the catalytic subunit Pol3, the structural B subunit Pol31, and Pol32, an additional auxiliary subunit. Pol3 interacts with Pol31 thanks to its C-terminal domain (CTD) and this interaction is of functional importance both in DNA replication and DNA repair. Interestingly, deletion of the last four C-terminal Pol3 residues, LSKW, in the pol3-ct mutant does not affect DNA replication but leads to defects in homologous recombination and in break-induced replication (BIR) repair pathways. The defect associated with pol3-ct could result from a defective interaction between Pol δ and a protein involved in recombination. However, we show that the LSKW motif is required for the interaction between Pol3 C-terminal end and Pol31. This loss of interaction is relevant in vivo since we found that pol3-ct confers HU sensitivity on its own and synthetic lethality with a POL32 deletion. Moreover, pol3-ct shows genetic interactions, both suppression and synthetic lethality, with POL31 mutant alleles. Structural analyses indicate that the B subunit of Pol δ displays a major conserved region at its surface and that pol31 alleles interacting with pol3-ct, correspond to substitutions of Pol31 amino acids that are situated in this particular region. Superimposition of our Pol31 model on the 3D architecture of the phylogenetically related DNA polymerase α (Pol α) suggests that Pol3 CTD interacts with the conserved region of Pol31, thus providing a molecular basis to understand the defects associated with pol3-ct. Taken together, our data highlight a stringent dependence on Pol δ complex stability in DNA repair.
Published: 2010
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28. Identification of a duplicated V3 domain in NS5A associated with cirrhosis and hepatocellular carcinoma in HCV-1b patients

Author: Elisabeth André-Garnier, J. B. Bour, Jacques Izopet, Christophe Lemaire, S. Vallet, Adeline Pivert, Arielle R. Rosenberg, Pascale Trimoulet, Christopher Payan, E. Bouthry, Pascal Veillon, Sophie Alain, Dominique Bettinger, P. André, Alexandra Ducancelle, Catherine Gaudy-Graffin, Jean-Michel Pawlotsky, Françoise Stoll-Keller, Odile Petsaris, Cécile Henquell, Yazid Baazia, Bruno Pozzetto, Françoise Lunel-Fabiani, Florence Abravanel, Henia Saoudin, V. Thibault, Anne Signori-Schmuck, Sandrine Bertrais, Gisèle Lagathu, H. Le Guillou-Guillemette, Sylvie Larrat, S. Minjolle-Cha, Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM), SFR UA 4208 Interactions Cellulaires et Applications Thérapeutiques (ICAT), Université d'Angers (UA), CHU Limoges, Service de virologie [CHU Pitié-Salpêtrière], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Laboratoire de Virologie [Toulouse], CHU Toulouse [Toulouse], Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), CHU Clermont-Ferrand, Hôtel-Dieu de Nantes, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne (LUBEM), Université de Brest (UBO), CHU Dijon, Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), CHU Avicenne, Service de virologie et d'immunologie biologique, CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Laboratoire de Bactériologie de l'Hôpital de la Croix-Rousse, Hospices Civils de Lyon (HCL), Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH - U1259 Inserm - CHRU Tours ), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), Biologie structurale des interactions entre virus et cellule hôte (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire de Saint-Etienne (CHU de Saint-Etienne), CHU Pontchaillou [Rennes], Interaction virus-hôte et maladies du foie, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Henri Mondor, The authors thank A. Lefebvre, E. Muel, Y. Desille, I. Renard, A. Plages, A. Rodallec and W. Merrouche for their assistance in clinical data recording.Funding: This study was funded by the ANRS (Agence de Nationale de Recherche sur le VIH et les hépatites, appel d’offre Mars 2006)., PRES Université Nantes Angers Le Mans [UNAM], Interactions cellulaires et applications thérapeutiques (ICAT), Université d'Angers (UA) - CHU Angers - CRLCC Paul Papin - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Service de virologie, Université Pierre et Marie Curie - Paris 6 (UPMC) - Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Pitié-Salpêtrière [APHP], Centre de Physiopathologie Toulouse Purpan, Université Paul Sabatier - Toulouse 3 (UPS) - IFR30 - IFR150 - Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Cochin [APHP], Université européenne de Bretagne (UEB) - Université de Bretagne Occidentale (UBO) - IFR148 ScInBioS, Centre Hospitalier Universitaire de Dijon (CHU Dijon), CHU Bordeaux [Bordeaux] - Groupe hospitalier Pellegrin, Hospices Civils de Lyon, Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH), Institut National de la Santé et de la Recherche Médicale (INSERM) - Université François Rabelais - Tours - CHRU Tours, CHU Besançon, Université de Franche-Comté (UFC), Laboratoire de biologie structurale des interactions entre virus et cellule hôte, Université Joseph Fourier - Grenoble 1 (UJF) - Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg - Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire Virologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire de Saint-Etienne [CHU Saint-Etienne] (CHU ST-E), CHU Henri Mondor [Créteil], and Jonchère, Laurent
Subjects: Adult, Liver Cirrhosis, Male, hepatitis C virus, Cirrhosis, Carcinoma, Hepatocellular, Hepatitis C virus, [SDV]Life Sciences [q-bio], Viral quasispecies, Hepacivirus, Biology, Viral Nonstructural Proteins, medicine.disease_cause, NS5A, Liver disease, Virology, Gene Duplication, Gene duplication, medicine, Prevalence, Humans, Aged, Hepatitis, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Sequence Analysis, RNA, cirrhosis, Liver Neoplasms, Hepatitis C, hepatocellular carcinoma, Hepatitis C, Chronic, Middle Aged, medicine.disease, digestive system diseases, 3. Good health, Protein Structure, Tertiary, [SDV] Life Sciences [q-bio], Mutagenesis, Insertional, Infectious Diseases, Cross-Sectional Studies, Hepatocellular carcinoma, RNA, Viral, Female, V3 domain, France, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; Background The NS5A protein of the hepatitis C virus has been shown to be involved in the development of hepatocellular carcinoma. Objectives In a French multicenter study, we investigated the clinical and epidemiological features of a new HCV genotype 1b strain bearing a wide insertion into the V3 domain. Study Design We studied NS5A gene sequences in 821 French patients infected with genotype 1b HCV. Results We identified an uncharacterized V3 insertion without ORF disruption in 3.05% of the HCV sequences. The insertion comprised 31 amino-acids for the majority of patients; 3 patients had 27 amino-acids insertions and 1 had a 12 amino-acids insertion. Sequence identity between the 31 amino-acids insertions and the V3 domain ranged from 48 to 96% with E-values above 4e-5, thus illustrating sequence homology and a partial gene duplication event that to our knowledge has never been reported in HCV. Moreover we showed the presence of the duplication at the time of infection and its persistence at least during 12 years in the entire quasispecies. No association was found with extrahepatic diseases. Conversely, patients with cirrhosis were two times more likely to have HCV with this genetic characteristic (p=0.04). Moreover, its prevalence increased with liver disease severity (from 3.0% in patients without cirrhosis to 9.4% in patients with both cirrhosis and HCC, p for trend=0.045). Conclusions We identified a duplicated V3 domain in the HCV-1b NS5A protein for the first time. The duplication may be associated with unfavorable evolution of liver disease including a possible involvement in liver carcinogenesis
Published: 2015
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29. Trm112, a hub protein interacting with and activating four methyltransferases involved in mRNA translation

Author: Graille, Marc, GRAILLE, Marc, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2015

30. Crystallization and preliminary X-ray crystallographic studies of a complex between theLactococcus lactisFpg DNA-repair enzyme and an abasic site containing DNA

Author: Charles Zelwer, Bertrand Castaing, Karine Pereira de Jésus, Laurence Serre, Nadège Hervouet, Véronique Bouckson-Castaing, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and LEGOUPIL, Laëtitia
Subjects: DNA repair, [SDV]Life Sciences [q-bio], Carbon-Oxygen Lyases, Biology, Crystallography, X-Ray, law.invention, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Structural Biology, law, DNA-(Apurinic or Apyrimidinic Site) Lyase, AP site, Crystallization, Furans, N-Glycosyl Hydrolases, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, DNA ligase, DNA clamp, Oligonucleotide, DNA, General Medicine, Formamidopyrimidine DNA glycosylase, Lactococcus lactis, [SDV] Life Sciences [q-bio], Crystallography, DNA-Formamidopyrimidine Glycosylase, chemistry, Propylene Glycols, 030220 oncology & carcinogenesis
Abstract: International audience; For protein-DNA complex crystallization, the choice of the DNA fragment is crucial. With the aim of crystallizing the 31 kDa Fpg DNA-repair enzyme bound to DNA, oligonucleotide duplexes varying in length, sequence, end type and nature of the specific DNA target site were used. Crystals of several protein-DNA combinations grew from solutions containing both polyethylene glycol and salt. This systematic crystallization screening followed by optimization of the crystallization conditions by microseeding led to crystals of Fpg bound to a 13 base-pair duplex DNA carrying the 1,3-propanediol abasic site analogue which are suitable for crystallographic analysis. Complete native data sets have been collected to 2.1 A resolution.
Published: 2002
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31. Pontin is a critical regulator for AML1-ETO-induced leukemia

Author: Breig, M, Bras, M., Martinez Soria, M, Osman, M., Heidenreich, O, Haenlin, M., Waltzer, L, Breig, O., Bras, S, Osman, O, Cognition, Langues, Langage, Ergonomie (CLLE-ERSS), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Toulouse - Jean Jaurès (UT2J)-Université Bordeaux Montaigne-Centre National de la Recherche Scientifique (CNRS), Cooperative Institute for Mesoscale Meteorological Studies (CIMMS), Laboratoire de Biologie structurale, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon
Subjects: Male, Cancer Research, Oncogene Proteins, Fusion, Chromosomes, Human, Pair 21, [SDV]Life Sciences [q-bio], Blotting, Western, Biology, Real-Time Polymerase Chain Reaction, Translocation, Genetic, RUNX1 Translocation Partner 1 Protein, RNA interference, hemic and lymphatic diseases, Biomarkers, Tumor, Tumor Cells, Cultured, medicine, Animals, Humans, RNA, Messenger, RNA, Small Interfering, Transcription factor, neoplasms, Cell Proliferation, Oligonucleotide Array Sequence Analysis, Regulation of gene expression, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Cell Cycle, DNA Helicases, Myeloid leukemia, Hematology, Cell cycle, medicine.disease, Molecular biology, Fusion protein, 3. Good health, Cell biology, Gene Expression Regulation, Neoplastic, Leukemia, Myeloid, Acute, Leukemia, Drosophila melanogaster, Oncology, Core Binding Factor Alpha 2 Subunit, ATPases Associated with Diverse Cellular Activities, Female, Carrier Proteins, Chromosomes, Human, Pair 8
Abstract: International audience; The oncogenic fusion protein AML1-ETO, also known as RUNX1-RUNX1T1 is generated by the t(8;21)(q22;q22) translocation, one of the most frequent chromosomal rearrangements in acute myeloid leukemia (AML). Identifying the genes that cooperate with or are required for the oncogenic activity of this chimeric transcription factor remains a major challenge. Our previous studies showed that Drosophila provides a genuine model to study how AML1-ETO promotes leukemia. Here, using an in vivo RNA interference screen for suppressors of AML1-ETO activity, we identified pontin/RUVBL1 as a gene required for AML1-ETO-induced lethality and blood cell proliferation in Drosophila. We further show that PONTIN inhibition strongly impaired the growth of human t(8;21)(+) or AML1-ETO-expressing leukemic blood cells. Interestingly, AML1-ETO promoted the transcription of PONTIN. Moreover, transcriptome analysis in Kasumi-1 cells revealed a strong correlation between PONTIN and AML1-ETO gene signatures and demonstrated that PONTIN chiefly regulated the expression of genes implicated in cell cycle progression. Concordantly, PONTIN depletion inhibited leukemic self-renewal and caused cell cycle arrest. All together our data suggest that the upregulation of PONTIN by AML1-ETO participate in the oncogenic growth of t(8;21) cells.
Published: 2014
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32. Structure-function analysis of the Trm9-Trm112 complex, a methyltransferase involved in the mcm5U34 tRNA modification and in response to genotoxic stresses

Author: Graille, Marc, GRAILLE, Marc, Laboratoire de Biologie Structurale de la Cellule (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2014

33. Effect of Rap1 binding on DNA distortion and potassium permanganate hypersensitivity

Author: Eric Gilson, Marie-Josèphe Giraud-Panis, Simona Miron, Eric Le Cam, Olivier Piétrement, Bianca Sclavi, B. Matot, Yann-Vaï Le Bihan, Marie-Hélène Le Du, Sylvaine Gasparini, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Myologie, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université de Reims Champagne-Ardenne (URCA), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Laboratoire de Biologie Moléculaire de la Cellule (LBMC), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Structurale et Radiobiologie (LBSR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Recherche sur le Cancer et le Vieillissement (IRCAN), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Laboratoire de biologie et pharmacologie appliquée (LBPA), Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Ecole Normale Supérieure Paris-Saclay (ENS Paris Saclay), Imagerie intégrative de la molécule à l'organisme, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bioénergétique, Biologie Stucturale, et Mécanismes (SB2SM), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Le Cam, Eric, Université Nice Sophia Antipolis (... - 2019) (UNS), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Centre National de la Recherche Scientifique (CNRS)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Sorbonne Université (SU), Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL)
Subjects: endocrine system, Saccharomyces cerevisiae Proteins, [SDV]Life Sciences [q-bio], Telomere-Binding Proteins, Mutant, Saccharomyces cerevisiae, Arginine, Crystallography, X-Ray, Shelterin Complex, Cytosine, 03 medical and health sciences, chemistry.chemical_compound, Nucleic acid thermodynamics, 0302 clinical medicine, Potassium Permanganate, Structural Biology, Binding site, DNA, Fungal, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Permanganate, Hydrogen Bonding, General Medicine, biology.organism_classification, Solutions, [SDV] Life Sciences [q-bio], enzymes and coenzymes (carbohydrates), Potassium permanganate, chemistry, Biochemistry, Biophysics, Nucleic Acid Conformation, 030217 neurology & neurosurgery, DNA, Protein Binding, Transcription Factors
Abstract: Repressor activator protein 1 (Rap1) is an essential factor involved in transcription and telomere stability in the budding yeast Saccharomyces cerevisiae. Its interaction with DNA causes hypersensitivity to potassium permanganate, suggesting local DNA melting and/or distortion. In this study, various Rap1-DNA crystal forms were obtained using specifically designed crystal screens. Analysis of the DNA conformation showed that its distortion was not sufficient to explain the permanganate reactivity. However, anomalous data collected at the Mn edge using a Rap1-DNA crystal soaked in potassium permanganate solution indicated that the DNA conformation in the crystal was compatible with interaction with permanganate ions. Sequence-conservation analysis revealed that double-Myb-containing Rap1 proteins all carry a fully conserved Arg580 at a position that may favour interaction with permanganate ions, although it is not involved in the hypersensitive cytosine distortion. Permanganate reactivity assays with wild-type Rap1 and the Rap1[R580A] mutant demonstrated that Arg580 is essential for hypersensitivity. AFM experiments showed that wild-type Rap1 and the Rap1[R580A] mutant interact with DNA over 16 successive binding sites, leading to local DNA stiffening but not to accumulation of the observed local distortion. Therefore, Rap1 may cause permanganate hypersensitivity of DNA by forming a pocket between the reactive cytosine and Arg580, driving the permanganate ion towards the C5-C6 bond of the cytosine.
Published: 2013
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34. Self-organization of the vesicular stomatitis virus nucleocapsid into a bullet shape

Author: Danielle Blondel, Irina Gutsche, Marc Jamin, Delphine Guilligay, Rob W.H. Ruigrok, Euripedes A. Ribeiro, Ambroise Desfosses, Guy Schoehn, Biologie Structurale des interactions entre Virus et Cellule Hôte, Laboratoire de biologie structurale des interactions entre virus et cellule hôte, Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), FCTUC/IT, Department of Electrical and Computer Engineering, University of Coimbra [Portugal] (UC), Institut de biologie structurale (IBS - UMR 5075), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS), Virologie moléculaire et structurale (VMS), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Unit of Virus Host Cell Interactions (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Biologie structurale des interactions entre virus et cellule hôte (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), and Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG)
Subjects: Multidisciplinary, Viral matrix protein, viruses, [SDV]Life Sciences [q-bio], Cryoelectron Microscopy, General Physics and Astronomy, General Chemistry, Biology, biology.organism_classification, Virology, General Biochemistry, Genetics and Molecular Biology, Vesicular stomatitis Indiana virus, Folding (chemistry), Viral Matrix Proteins, Nucleoproteins, Vesicular stomatitis virus, Biophysics, Nucleic Acid Conformation, RNA, Viral, Nucleocapsid, Bullet (shape)
Abstract: International audience; The typical bullet shape of Rhabdoviruses is thought to rely on the matrix protein for stabilizing the nucleocapsid coil. Here we scrutinize the morphology of purified and recombinant nucleocapsids of vesicular stomatitis virus in vitro. We elucidate pH and ionic strength conditions for their folding into conical tips and further growth into whole bullets, and provide cryo-electron microscopy reconstructions of the bullet tip and the helical trunk. We address conformational variability of the reconstituted nucleocapsids and the issue of constraints imposed by the binding of matrix protein. Our findings bridge the gap between the isolated nucleoprotein-RNA string in its form of an undulating ribbon, and the tight bullet-shaped virion skeleton.
Published: 2013
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35. Myeloid leukemia factor is a conserved regulator of RUNX transcription factor activity involved in hematopoiesis

Author: Bras, Stéphanie, Martin-Lannerée, Séverine, Gobert, Vanessa, Breig, Osman, Sanial, Matthieu, Yamaguchi, Masamitsu, Haenlin, Marc, Plessis, Anne, Waltzer, Lucas, Martin-Lanneree, S., Augé, Benoit, Toxicité environnementale, cibles thérapeutiques, signalisation cellulaire (T3S - UMR_S 1124), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biotechnologies Végétales appliquées aux Plantes Aromatiques et Médicinales (LBVPAM), Université Jean Monnet [Saint-Étienne] (UJM), Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), National Institute for Materials Science (NIMS), Biologie moléculaire et génie génétique (IGBMC), Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de biologie du développement (CBD), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Institut d'astrophysique spatiale (IAS), Université Paris-Sud - Paris 11 (UP11)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie structurale, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Transcriptional Activation, Embryo, Nonmammalian, Oncogene Proteins, Fusion, [SDV]Life Sciences [q-bio], Biology, Blood cell, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, hemic and lymphatic diseases, medicine, Animals, Drosophila Proteins, Humans, Cell Lineage, Transcription factor, Conserved Sequence, 030304 developmental biology, 0303 health sciences, Multidisciplinary, Protein Stability, Myeloid leukemia, Core Binding Factor alpha Subunits, Biological Sciences, medicine.disease, Fusion protein, Molecular biology, Cell biology, Hematopoiesis, DNA-Binding Proteins, Haematopoiesis, Leukemia, medicine.anatomical_structure, Drosophila melanogaster, Phenotype, RUNX1, chemistry, 030220 oncology & carcinogenesis, Larva, Core Binding Factor Alpha 2 Subunit, Proteolysis, Drosophila Protein, Transcription Factors
Abstract: Defining the function of the genes that, like RUNX1 , are deregulated in blood cell malignancies represents an important challenge. Myeloid leukemia factors (MLFs) constitute a poorly characterized family of conserved proteins whose founding member, MLF1, has been associated with acute myeloid leukemia in humans. To gain insight into the functions of this family, we investigated the role of the Drosophila MLF homolog during blood cell development. Here we report that mlf controls the homeostasis of the Drosophila hematopoietic system. Notably, mlf participates in a positive feedback loop to fine tune the activity of the RUNX transcription factor Lozenge (LZ) during development of the crystal cells, one of the two main blood cell lineages in Drosophila . At the molecular level, our data in cell cultures and in vivo strongly suggest that MLF controls the number of crystal cells by protecting LZ from degradation. Remarkably, it appears that the human MLF1 protein can substitute for MLF in the crystal cell lineage. In addition, MLF stabilizes the human oncogenic fusion protein RUNX1-ETO and is required for RUNX1-ETO–induced blood cell disorders in a Drosophila model of leukemia. Finally, using the human leukemic blood cell line Kasumi-1, we show that MLF1 depletion impairs RUNX1-ETO accumulation and reduces RUNX1-ETO–dependent proliferation. Thus, we propose that the regulation of RUNX protein levels is a conserved feature of MLF family members that could be critical for normal and pathological blood cell development.
Published: 2012
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36. Cyclic GMP-dependent protein kinase potentiates serotonin-induced Egr-1 binding activity in PC12 cells

Author: Pierre G. Lutz, Jean Zwiller, Nathalie Thiriet, Laurent Esteve, Dominique Aunis, Marie-Odile Revel, Biologie de la communication cellulaire, IFR 31 Louis Bugnard (IFR 31), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie structurale, Institut National de la Santé et de la Recherche Médicale (INSERM), Lutz, Pierre, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: MAPK/ERK pathway, Time Factors, 8-bromo-cGMP, [SDV]Life Sciences [q-bio], 5-HT, Mitogen-activated protein kinase kinase, PC12 Cells, 0302 clinical medicine, ASK1, Cloning, Molecular, Enzyme Inhibitors, Phosphorylation, 0303 health sciences, Chemistry, PKG, MEK, Up-Regulation, Cell biology, [SDV] Life Sciences [q-bio], DNA-Binding Proteins, 8pCPT-cGMP, cGMP-dependent protein kinase, cardiovascular system, Mitogen-Activated Protein Kinases, Signal transduction, Protein Binding, Signal Transduction, c-fos, Serotonin, DNA, Complementary, Immunoblotting, 8Br-cGMP, Transfection, 8-p-chlorophenylthio-cGMP, Immediate early protein, Immediate-Early Proteins, 03 medical and health sciences, ERK MAP kinase, cyclic GMP, MBP, Cyclic GMP-Dependent Protein Kinases, Animals, Humans, Protein kinase A, Early Growth Response Protein 1, 030304 developmental biology, Cell Nucleus, Mitogen-Activated Protein Kinase Kinases, MAP kinase kinase kinase, egr-1, PC12 cells cGMP, Cell Biology, Blotting, Northern, myelin basic protein, Molecular biology, Rats, cGMP, Egr-1 transcription factor, MAP or ERK kinase, 5-hydroxytryptamine or serotonin, 030217 neurology & neurosurgery, Transcription Factors
Abstract: The NO/cyclic GMP (cGMP) signal transduction pathway, which involves the cGMP-dependent protein kinase (PKG), regulates transcription of several genes, including immediate early genes. Using transfection experiments with the PKG-Ialpha cDNA cloned from human aorta, we show here that addition of membrane-permeable cGMP analogues to PC12 cells slightly upregulated ERK MAP (mitogen-activated protein) kinase. Likewise, PKG-Ialpha was found to activate weakly DNA binding activity of the Egr-1 transcription factor. On the other hand, PKG-Ialpha overexpression was shown to tremendously amplify the Egr-1 binding activity induced by the neurotransmitter serotonin, which activates egr-1 gene expression also via the stimulation of the ERK MAP kinase pathway. Since this potentiation occurred neither at the level of ERK nor at the egr-1 transcriptional level, the mechanism of amplification probably results from the convergence of ERK and PKG pathways at the level of the transcription factor Egr-1.
Published: 2001

37. Cleavage and binding of a DNA fragment containing a single 8-oxoguanine by wild type and mutant FPG proteins

Author: Jacques Laval, Peter Nehls, Charles Zelwer, Bertrand Castaing, Serge Boiteux, Hartmut Seliger, Albert Geiger, LEGOUPIL, Laëtitia, Laboratoire de Biologie Structurale [Gif sur Yvette], Centre National de la Recherche Scientifique (CNRS), Sektion Polymere [Ulm, Germany], Universität Ulm - Ulm University [Ulm, Allemagne], Lab. für Molekularbiologie and Allgemeine Pathologie [Munich, Germany], Groupe Reparation des Lesions Radio- et Chimio-induites [Villejuif], and Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Guanine, endocrine system diseases, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Mutant, Glycine, Biology, Substrate Specificity, chemistry.chemical_compound, Escherichia coli, Genetics, heterocyclic compounds, Amino Acid Sequence, Cysteine, N-Glycosyl Hydrolases, Peptide sequence, Base Composition, Base Sequence, Oligonucleotide, Escherichia coli Proteins, Wild type, nutritional and metabolic diseases, Zinc Fingers, DNA, Molecular biology, 8-Oxoguanine, [SDV] Life Sciences [q-bio], Kinetics, DNA-Formamidopyrimidine Glycosylase, chemistry, Mutagenesis, Duplex (building)
Abstract: International audience; A 34-mer oligonucleotide containing a single 7,8-dihydro-8-oxoguanine (8-OxoG) residue was used to study the enzymatic and DNA binding properties of the Fpg protein from E. coli. The highest rates of incision of the 8-OxoG containing strand by the Fpg protein were observed for duplexes where 8-OxoG was opposite C (*G/C) or T (*G/T). In contrast, the rates of incision of duplexes containing 8-OxoG opposite G (*G/G) and A (*G/A) were 5-fold and 200-fold slower. Gel retardation studies showed that the Fpg protein had a strong affinity for duplexes where the 8-OxoG was opposite pyrimidines and less affinity for duplexes where the 8-OxoG was opposite purines. KDapp values were 0.6 nM (*G/C), 1.0 nM (*G/T), 6.0 nM (*G/G) and 16.0 nM (*G/A). The Fpg protein also binds to unmodified (G/C) duplex and a KDapp of 90 nM was measured. The cleavage and binding of the (*G/C) duplex were also studied using bacterial crude lysates. Wild type E. coli crude extract incised the 8-OxoG containing strand and formed a specific retardation complex with the (*G/C) duplex. These two reactions were mediated by the Fpg protein, since they were not observed with a crude extract from a bacterial strain whose fpg gene was inactivated. Furthermore, we have studied the properties of 6 mutant Fpg proteins with Cys-->Gly mutations. The results showed that the 2 Fpg proteins with Cys-->Gly mutations outside the zinc finger sequence cleaved the 8-OxoG containing strand, formed complexes with the (*G/C) duplex and suppressed the mutator phenotype of the fpg-1 mutant. In contrast, the 4 Fpg proteins with Cys-->Gly mutations within the zinc finger motif neither cleave nor bind the (*G/C) duplex, nor do these proteins suppress the fpg-1 mutator phenotype.
Published: 1993
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38. Specificity, T cell receptor diversity and activation requirements of CD4+ and CD8+ clones derived from human melanoma-infiltrating lymphocytes

Author: Marie-Christine Pandolfino, Nadine Gervois, Christophe Viret, Elisabeth Diez, François Davodeau, Yannick Guilloux, Francine Jotereau, Recherche sur les effecteurs lymphocytaires T, Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie structurale, and GERVOIS, Nadine
Subjects: CD8 Antigens, [SDV]Life Sciences [q-bio], Immunology, Receptors, Antigen, T-Cell, chemical and pharmacologic phenomena, Recombinant Interleukin, Biology, Lymphocyte Activation, Major histocompatibility complex, 03 medical and health sciences, Lymphocytes, Tumor-Infiltrating, 0302 clinical medicine, Antigen, Tumor Cells, Cultured, Humans, Immunology and Allergy, Melanoma, 030304 developmental biology, 0303 health sciences, Tumor-infiltrating lymphocytes, Histocompatibility Antigens Class I, T-cell receptor, Histocompatibility Antigens Class II, T lymphocyte, Autologous tumor cell, Molecular biology, Clone Cells, [SDV] Life Sciences [q-bio], 030220 oncology & carcinogenesis, CD4 Antigens, biology.protein, Interleukin-2, CD8
Abstract: International audience; To try to understand the functional significance of human melanoma-infiltrating lymphocytes (TIL), a clonal analysis of the specificity, T cell receptor (TcR) diversity and activation requirements of these lymphocytes isolated from four different tumors was carried out. Supporting the presence of in vivo primed tumor-specific T lymphocytes in these four tumors, a high frequency of the Cd8+ and CD4+ clones, obtained from the TIL cultured for a few days with recombinant interleukin (rIL)-2 and autologous tumor cells, exhibited a restricted lysis or proliferation in response to the autologous tumor cell line. In contrast, no tumor-specific clone was obtained from freshly extracted TIL, suggesting that the frequency of tumor-specific effectors remained low in these tumors. Only the CD8+ clones lysed the autologous tumor cells and their activity was major histocompatibility complex MHC class I restricted. Significant expansion of CD4+ and CD8+ tumor-specific clones required regular restimulation by autologous melanoma cells but also the addition of exogenous IL-2 and of Epstein-Barr virus-transformed B feeder cells. Five different tumor-specific clones, three CD8+ and two CD4+ clones were identified in a single tumor on the basis of their TcR gene configuration. Together, these data suggest that a spontaneous and diverse immune response, mediated by tumor-specific CD4+ as well as CD8+ T lymphocytes, arises in most MHC-bearing human melanomas but that antigen-MHC complex presentation by tumor cells does not, at least in vitro, allow a significant proliferation of these lymphocytes.
Published: 1992
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39. Local structure of a peptide contact site on Ak alpha

Author: Nadine Gervois, Christophe Benoist, Diane Mathis, Luciano Adorini, Bing Yuan Wei, Georges Mer, Laboratoire de Biologie structurale, Institut National de la Santé et de la Recherche Médicale (INSERM), Sandoz Pharma Ltd., and GERVOIS, Nadine
Subjects: structure/function, T-Lymphocytes, T cell, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Immunology, Mutant, Receptors, Antigen, T-Cell, Antigen-Presenting Cells, Peptide binding, Peptide, Cell Line, Mice, Structure-Activity Relationship, 03 medical and health sciences, L Cells, 0302 clinical medicine, Complementary DNA, medicine, Animals, Immunology and Allergy, Ribonuclease, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Binding Sites, Hybridomas, Base Sequence, biology, Histocompatibility Antigens Class II, General Medicine, Amino acid, [SDV] Life Sciences [q-bio], antigen presentation, medicine.anatomical_structure, Biochemistry, chemistry, biology.protein, MHC class II, Alpha chain, mutagenesis, 030215 immunology
Abstract: International audience; We have sought to determine how much amino acid diversity is tolerable at position 69 of the A* chain, a position previously implicated as a peptlde contact site. Slot-machine mutagenesis was used to create a set of 11 mutant A* cDNAs, each specifying a different amino acid at position 69. These cDNAs were individually expressed In L cells together with a wild-type AJj cDNA to produce a panel of mutant antigen-presenting cell lines. The ability of each member of this panel to present a hen egg lysozyme and a bovine ribonuciease peptlde to various T hybrldomas was assessed. We found that a surprising degree of amino acid diversity is tolerable at A* position 69: even charged (Glu, Arg) or bulky (Trp, Tyr) residues can be accommodated without abrogating cell-surface expression of A k , peptide binding to it, or T cell recognition of It. We discuss the Implications of these findings for models of T cell recognition of the class II molecule/antigen duplex.
Published: 1991
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40. Changes in the chromatin structure of Drosophila glue genes accompany developmental cessation of transcription in wild type and transformed strains

Author: Adam Sobkowiak, Marc Bourouis, Guy Dretzen, Maria Bellard, Geoff Richards, Philippe Ramain, Laboratoire de Génetique Moléculaire des Eucaryotes (LGME), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie structurale, Institut National de la Santé et de la Recherche Médicale (INSERM), Physiopathologie du système nerveux., Université Louis Pasteur - Strasbourg I-IFR37-Institut National de la Santé et de la Recherche Médicale (INSERM), and RAMAIN, Philippe
Subjects: Transcription, Genetic, [SDV]Life Sciences [q-bio], Biology, General Biochemistry, Genetics and Molecular Biology, Salivary Glands, Cell Line, chemistry.chemical_compound, Transcription (biology), medicine, Animals, Salivary Proteins and Peptides, Gene, Deoxyribonucleases, Salivary gland, Wild type, Molecular biology, Chromatin, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, Drosophila melanogaster, chemistry, Gene Expression Regulation, Genes, Larva, DNA
Abstract: Three Drosophila salivary gland glue genes show a dramatic transition in their DNAase I hypersensitive sites during the short period between the late third instar and the white prepupa, which correlates with the cessation of their transcription. In culture cells, where the genes are inactive, there is a chromatin configuration similar to that of prepupal salivary glands. In two transformed fly strains where the sgs3 gene is active at new chromosomal sites, including one in which 2.6 kb of sgs3 upstream sequences have been inverted, the same DNAase I hypersensitive sites and developmental transitions are seen over the same DNA regions. These results, together with the analysis of transformants carrying rearranged sgs3 genes, suggest that there is at least one distal DNAase I hypersensitive sensitive site associated with an element of regulation which may be exchanged between sgs genes.
Published: 1986
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