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19 results on '"MESH : Cell Nucleus"'

1. BAT3 modulates p300-dependent acetylation of p53 and autophagy-related protein 7 (ATG7) during autophagy

Author

Laetitia K. Linares, Andrew V. Hubberstey, Emmanuelle Liaudet-Coopman, Nelly Pirot, Fabienne Desmots, Guy Berchem, Anne-Sophie Bach, Esther Pérez-Gracia, Christine Prébois, Valérie Palissot, Patrice Codogno, Céline Gongora, Salwa Sebti, Sophie Pattingre, Chantal Bauvy, Institut de recherche en cancérologie de Montpellier ( IRCM - U896 Inserm - UM1 ), Université Montpellier 1 ( UM1 ) -CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Institut Necker Enfants-Malades (INEM) ( INEM - UM 111 (UMR 8253 / U1151) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Service d'Hématologie, Immunologie et de Thérapie Cellulaire ( HITC ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Department of Biological Sciences [Windsor], University of Windsor [Ca], Laboratory of Experimental Hemato-Oncology ( CRP-Santé ), Centre de Recherche Public-Santé, This work is supported by INSERM, ARC (SP), La Ligue Régionale contre Le Cancer (Comités du Gard et de l'Hérault) (SP), Cancéropole GSO (SP) and CHEMORES consortium (EU FP6, ELC). SS has fellowships from the FNR Luxembourg and La Ligue Nationale contre le Cancer., Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), CRLCC Val d'Aurelle - Paul Lamarque-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Hématologie, Immunologie et de Thérapie Cellulaire (HITC), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Laboratory of Experimental Hemato-Oncology (CRP-Santé), Le Ster, Yves, Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), and Université de Rennes (UR)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes]

Subjects
p53, Autophagy-Related Protein 7, MESH: Mice, Knockout, Mice, Cytosol, MESH: Cytosol, MESH : Embryo, Mammalian, MESH: Animals, Nuclear protein, MESH: Tumor Suppressor Protein p53, Mice, Knockout, MESH : Cell Nucleus, Multidisciplinary, MESH: Real-Time Polymerase Chain Reaction, MESH : Cytosol, Life Sciences, Nuclear Proteins, Biological Sciences, Cell biology, medicine.anatomical_structure, MESH : Cell Fractionation, MESH: Cell Fractionation, MESH : DNA Primers, MESH: Molecular Chaperones, Microtubule-Associated Proteins, MESH: Acetylation, MESH: Cell Nucleus, MESH: DNA Primers, MESH : Real-Time Polymerase Chain Reaction, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Cell Fractionation, Real-Time Polymerase Chain Reaction, MESH : Acetylation, BAG3, MESH : E1A-Associated p300 Protein, MESH : Immunoprecipitation, BAT3, MESH : Mice, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Autophagy, medicine, Animals, Immunoprecipitation, MESH: Autophagy, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MESH: Mice, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, DNA Primers, acetylation, Cell Nucleus, MESH: Immunoprecipitation, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Embryo, Mammalian, MESH : Molecular Chaperones, MESH : Nuclear Proteins, Embryo, Mammalian, MESH : Autophagy, Molecular biology, ATG, MESH: Microtubule-Associated Proteins, MESH : Tumor Suppressor Protein p53, Cell nucleus, MESH : Microtubule-Associated Proteins, Acetylation, MESH : Mice, Knockout, MESH: E1A-Associated p300 Protein, MESH : Animals, Tumor Suppressor Protein p53, E1A-Associated p300 Protein, MESH: Nuclear Proteins, Nuclear localization sequence, Molecular Chaperones
Abstract

International audience; Autophagy is regulated by posttranslational modifications, including acetylation. Here we show that HLA-B-associated transcript 3 (BAT3) is essential for basal and starvation-induced autophagy in embryonic day 18.5 BAT3(-/-) mouse embryos and in mouse embryonic fibroblasts (MEFs) through the modulation of p300-dependent acetylation of p53 and ATG7. Specifically, BAT3 increases p53 acetylation and proautophagic p53 target gene expression, while limiting p300-dependent acetylation of ATG7, a mechanism known to inhibit autophagy. In the absence of BAT3 or when BAT3 is located exclusively in the cytosol, autophagy is abrogated, ATG7 is hyperacetylated, p53 acetylation is abolished, and p300 accumulates in the cytosol, indicating that BAT3 regulates the nuclear localization of p300. In addition, the interaction between BAT3 and p300 is stronger in the cytosol than in the nucleus and, during starvation, the level of p300 decreases in the cytosol but increases in the nucleus only in the presence of BAT3. We conclude that BAT3 tightly controls autophagy by modulating p300 intracellular localization, affecting the accessibility of p300 to its substrates, p53 and ATG7.

Published
2014

2. Trm112p Is a 15-kDa Zinc Finger Protein Essential for the Activity of Two tRNA and One Protein Methyltransferases in Yeast

Author

Léon Dirick, Bruno Lapeyre, Glenn R. Björk, Marie-Hélène Mazauric, Suresh K. Purushothaman, Centre de recherches de biochimie macromoléculaire ( CRBM ), Université Montpellier 1 ( UM1 ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -IFR122-Centre National de la Recherche Scientifique ( CNRS ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de recherche en Biologie Cellulaire (CRBM), Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1), Institut de Génétique Moléculaire de Montpellier (IGMM), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)

Subjects
Proteomics, MESH : DNA, MESH: tRNA Methyltransferases, MESH : Saccharomyces cerevisiae, MESH : Models, Genetic, Biochemistry, MESH: Recombinant Proteins, MESH : Proteomics, MESH: Saccharomyces cerevisiae Proteins, MESH : Anticodon, RNA, Transfer, MESH : Catalysis, MESH: Models, Genetic, Zinc finger, tRNA Methyltransferases, MESH : Cell Nucleus, 0303 health sciences, MESH : Chromatin, MESH: Proteomics, 030302 biochemistry & molecular biology, MESH: DNA, Zinc Fingers, MESH : Protein Binding, Translation (biology), MESH: Saccharomyces cerevisiae, MESH : Mitosis, Chromatin, Recombinant Proteins, MESH : tRNA Methyltransferases, Transfer RNA, T arm, MESH : Mutation, Protein Binding, MESH: Cell Nucleus, TRNA modification, Saccharomyces cerevisiae Proteins, MESH: Mutation, MESH : Recombinant Proteins, Saccharomyces cerevisiae, Mitosis, MESH : RNA, Transfer, Biology, Catalysis, Chromatin remodeling, MESH: Chromatin, MESH : Saccharomyces cerevisiae Proteins, 03 medical and health sciences, Anticodon, MESH: Anticodon, MESH: Zinc Fingers, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Zinc Fingers, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, 030304 developmental biology, Cell Nucleus, Models, Genetic, TRNA Methyltransferase, DNA, Cell Biology, MESH: Mitosis, MESH: Catalysis, MESH: RNA, Transfer, biology.organism_classification, Mutation, RNA
Abstract

International audience; The degenerate base at position 34 of the tRNA anticodon is the target of numerous modification enzymes. In Saccharomyces cerevisiae, five tRNAs exhibit a complex modification of uridine 34 (mcm(5)U(34) and mcm(5)s(2)U(34)), the formation of which requires at least 25 different proteins. The addition of the last methyl group is catalyzed by the methyltransferase Trm9p. Trm9p interacts with Trm112p, a 15-kDa protein with a zinc finger domain. Trm112p is essential for the activity of Trm11p, another tRNA methyltransferase, and for Mtq2p, an enzyme that methylates the translation termination factor eRF1/Sup45. Here, we report that Trm112p is required in vivo for the formation of mcm(5)U(34) and mcm(5)s(2)U(34). When produced in Escherichia coli, Trm112p forms a complex with Trm9p, which renders the latter soluble. This recombinant complex catalyzes the formation of mcm(5)U(34) on tRNA in vitro but not mcm(5)s(2)U(34). An mtq2-0 trm9-0 strain exhibits a synthetic growth defect, thus revealing the existence of an unexpected link between tRNA anticodon modification and termination of translation. Trm112p is associated with other partners involved in ribosome biogenesis and chromatin remodeling, suggesting that it has additional roles in the cell.

Published
2010

3. EFD Is an ERF Transcription Factor Involved in the Control of Nodule Number and Differentiation inMedicago truncatula

Author

Françoise de Billy, Andreas Niebel, Tatiana Vernié, Christian Rogers, Florian Frugier, Giles E. D. Oldroyd, Julie Plet, Jean-Philippe Combier, Pascal Gamas, Sandra Moreau, Laboratoire des interactions plantes micro-organismes ( LIPMO ), Institut National de la Recherche Agronomique ( INRA ) -Centre National de la Recherche Scientifique ( CNRS ), Ecologie microbienne ( EM ), Centre National de la Recherche Scientifique ( CNRS ) -Ecole Nationale Vétérinaire de Lyon ( ENVL ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique ( INRA ) -VetAgro Sup ( VAS ), Institut des sciences du végétal ( ISV ), Centre National de la Recherche Scientifique ( CNRS ), Laboratoire des interactions plantes micro-organismes (LIPM), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Ecole Nationale Vétérinaire de Lyon (ENVL), Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Lyon (ENVL)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)

Subjects
MESH: Signal Transduction, 0106 biological sciences, [ SDV.BV ] Life Sciences [q-bio]/Vegetal Biology, Cytokinins, MESH : Molecular Sequence Data, MESH : Transcription Factors, MESH: Feedback, Physiological, Regulator, MESH : Gene Deletion, Plant Science, Plant Root Nodulation, 01 natural sciences, Nod factor, chemistry.chemical_compound, MESH : Nitrogen Fixation, MESH: Phylogeny, MESH: Medicago truncatula, Research Articles, Phylogeny, Plant Proteins, Regulator gene, Feedback, Physiological, MESH : Cell Nucleus, 0303 health sciences, MESH: Plant Proteins, food and beverages, MESH: Transcription Factors, MESH : Feedback, Physiological, MESH : Cytokinins, Medicago truncatula, Cell biology, MESH: Root Nodules, Plant, Multigene Family, Cytokinin, RNA Interference, MESH: Cytokinins, Root Nodules, Plant, MESH: Plant Root Nodulation, Signal Transduction, MESH: Cell Nucleus, MESH: Ethylenes, Molecular Sequence Data, MESH: RNA Interference, MESH : Multigene Family, Biology, MESH: Nitrogen Fixation, MESH: Gene Expression Profiling, MESH : Root Nodules, Plant, 03 medical and health sciences, Nitrogen Fixation, MESH : Plant Proteins, Botany, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Transcription factor, 030304 developmental biology, Cell Nucleus, MESH : Signal Transduction, MESH: Molecular Sequence Data, Gene Expression Profiling, MESH : Gene Expression Profiling, MESH : Medicago truncatula, fungi, MESH : Phylogeny, MESH : Sinorhizobium meliloti, Cell Biology, Ethylenes, Meristem, biology.organism_classification, Response regulator, chemistry, MESH: Gene Deletion, MESH: Multigene Family, MESH : RNA Interference, MESH: Sinorhizobium meliloti, MESH : Plant Root Nodulation, Gene Deletion, MESH : Ethylenes, Sinorhizobium meliloti, Transcription Factors, 010606 plant biology & botany
Abstract

Mechanisms regulating legume root nodule development are still poorly understood, and very few regulatory genes have been cloned and characterized. Here, we describe EFD (for ethylene response factor required for nodule differentiation), a gene that is upregulated during nodulation in Medicago truncatula. The EFD transcription factor belongs to the ethylene response factor (ERF) group V, which contains ERN1, 2, and 3, three ERFs involved in Nod factor signaling. The role of EFD in the regulation of nodulation was examined through the characterization of a null deletion mutant (efd-1), RNA interference, and overexpression studies. These studies revealed that EFD is a negative regulator of root nodulation and infection by Rhizobium and that EFD is required for the formation of functional nitrogen-fixing nodules. EFD appears to be involved in the plant and bacteroid differentiation processes taking place beneath the nodule meristem. We also showed that EFD activated Mt RR4, a cytokinin primary response gene that encodes a type-A response regulator. We propose that EFD induction of Mt RR4 leads to the inhibition of cytokinin signaling, with two consequences: the suppression of new nodule initiation and the activation of differentiation as cells leave the nodule meristem. Our work thus reveals a key regulator linking early and late stages of nodulation and suggests that the regulation of the cytokinin pathway is important both for nodule initiation and development.

Published
2008

4. A multi-locus inference of the evolutionary diversification of extant flamingos (Phoenicopteridae)

Author

Brittney Ferrari, Marcel van Tuinen, Lisa M. Ogawa, Christopher R. Torres, Mark A. F. Gillingham, Department of Biology and Marine Biology, University of North Carolina Wilmington, Biogéosciences [Dijon] ( BGS ), AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université de Bourgogne ( UB ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de recherche de la Tour du Valat, Department of Evolutionary Genetics, Leibniz Institute for Zoo and Wildlife Research, and Financial support from the College of Arts and Sciences and Department of Biology and Marine Biology at UNCW, the Conseil Régional de Bourgogne, and the Total Foundation.

Subjects
Bill, Biogeography, Population, MESH : Multilocus Sequence Typing, Zoology, Metapopulation, Mirandornithes, Birds, Evolution, Molecular, MESH : Bayes Theorem, Phylogenetics, MESH : Genetic Variation, MESH : Biological Evolution, MESH : Phylogeography, Animals, MESH : Evolution, Molecular, MESH : Birds, education, Divergence time, Ecology, Evolution, Behavior and Systematics, Phylogeny, Cell Nucleus, education.field_of_study, MESH : Cell Nucleus, biology, Phylogenetic tree, Fossils, Genetic Variation, Bayes Theorem, Fossil, biology.organism_classification, Biological Evolution, Phoenicoparrus, Phoenicopteridae, Phylogeography, Flamingo, Evolutionary biology, Filter feeding, MESH : Fossils, MESH : Animals, [ SDV.BID.SPT ] Life Sciences [q-bio]/Biodiversity/Systematics, Phylogenetics and taxonomy, Multilocus Sequence Typing, Research Article
Abstract

9 pages; International audience; BACKGROUND: Modern flamingos (Phoenicopteridae) occupy a highly specialized ecology unique among birds and represent a potentially powerful model system for informing the mechanisms by which a lineage of birds adapts and radiates. However, despite a rich fossil record and well-studied feeding morphology, molecular investigations of the evolutionary progression among modern flamingos have been limited. Here, using three mitochondrial (mtDNA) markers, we present the first DNA sequence-based study of population genetic variation in the widely distributed Chilean Flamingo and, using two mtDNA and 10 nuclear (nDNA) markers, recover the species tree and divergence time estimates for the six extant species of flamingos. Phylogenetic analyses include likelihood and Bayesian frameworks and account for potential gene tree discordance. Analyses of divergence times are fossil calibrated at the divergence of Mirandornithes (flamingos + grebes) and the divergence of crown grebes. RESULTS: mtDNA sequences confirmed the presence of a single metapopulation represented by two minimally varying mtDNA barcodes in Chilean flamingos. Likelihood and Bayesian methods recovered identical phylogenies with flamingos falling into shallow-keeled (comprising the Greater, American and Chilean Flamingos) and deep-keeled (comprising the Lesser, Andean and James's Flamingos) sub-clades. The initial divergence among flamingos occurred at or shortly after the Mio-Pliocene boundary (6-3 Ma) followed by quick consecutive divergences throughout the Plio-Pleistocene. There is significant incongruence between the ages recovered by the mtDNA and nDNA datasets, likely due to mutational saturation occurring in the mtDNA loci. CONCLUSION: The finding of a single metapopulation in the widespread Chilean Flamingo confirms similar findings in other widespread flamingo species. The robust species phylogeny is congruent with previous classifications of flamingos based on feeding morphology. Modern phoenicopterids likely originated in the New World with each sub-clade dispersing across the Atlantic at least once. Our divergence time estimates place flamingos among the youngest families of birds, counter to the classical notion of flamingos as among the oldest based on biogeography and the fossil record. Finally, we designate 'Phoeniconaias' as a junior synonym of 'Phoenicoparrus' and redefine the latter genus as containing all flamingos more closely related to Phoenicoparrus andinus than Phoenicopterus roseus.

Published
2014

5. Ancient DNA suggests the leading role played by men in the Neolithic dissemination

Author

Lacan, Marie, Keyser, Christine, Ricaut, François-Xavier, Brucato, Nicolas, Tarrús, Josep, Bosch, Angel, Guilaine, Jean, Crubézy, Eric, Ludes, Bertrand, Tarrus, J., Crubezy, E., Laboratoire d'Anthropobiologie (LA), École des hautes études en sciences sociales (EHESS)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Anthropologie Moléculaire, Institut de Médecine Légale, Strasbourg, France (IML), Université Louis Pasteur - Strasbourg I, Anthropologie Moléculaire et Imagerie de Synthèse (AMIS), Université Toulouse III - Paul Sabatier (UT3), Collège de France (CdF (institution)), Institute of Legal Medicine, Université Paris Descartes - Paris 5 (UPD5), Centre National de la Recherche Scientifique (CNRS), Human Genetics, Collège de France (CDF), Collège de France (CdF), Institut de Médecine Légale - IML (Paris, France), Laboratoire d'Anthropobiologie ( LA ), École des hautes études en sciences sociales ( EHESS ) -Université Toulouse - Jean Jaurès ( UT2J ) -Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire d'Anthropologie Moléculaire, Institut de Médecine Légale, Strasbourg, France ( IML ), Anthropologie Moléculaire et Imagerie de Synthèse ( AMIS ), PRES Université de Toulouse-Centre National de la Recherche Scientifique ( CNRS ), Collège de France ( CDF ), Collège de France ( CdF ), and Université Paris Descartes - Paris 5 ( UPD5 )

Subjects
Male, MESH : DNA, [SHS.ANTHRO-BIO]Humanities and Social Sciences/Biological anthropology, MESH: History, Ancient, MESH: Fossils, Haplogroup, Dna genetics, Kinship, MESH : Chromosomes, Human, Y, MESH : History, Ancient, History, Ancient, MESH : Cell Nucleus, 0303 health sciences, Multidisciplinary, geography.geographical_feature_category, Fossils, 030305 genetics & heredity, MESH: DNA, Biological Sciences, humanities, Genealogy, Mitochondria, [ SDV.GEN.GPO ] Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE], Western europe, MESH : Mitochondria, Gene pool, MESH: Cell Nucleus, MESH: Mitochondria, MESH : Male, Biology, 03 medical and health sciences, MESH: Chromosomes, Human, Y, Cave, Humans, MESH: Role, MESH : Role, 030304 developmental biology, Cell Nucleus, geography, Chromosomes, Human, Y, MESH: Humans, [SDV.GEN.GPO]Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE], MESH : Humans, Role, DNA, MESH: Male, Ancient DNA, Evolutionary biology, [ SHS.ANTHRO-BIO ] Humanities and Social Sciences/Biological anthropology, MESH : Fossils, Biological dispersal
Abstract

International audience; The impact of the Neolithic dispersal on the western European populations is subject to continuing debate. To trace and date genetic lineages potentially brought during this transition and so understand the origin of the gene pool of current populations, we studied DNA extracted from human remains excavated in a Spanish funeral cave dating from the beginning of the fifth millennium B.C. Thanks to a "multimarkers" approach based on the analysis of mitochondrial and nuclear DNA (autosomes and Y-chromosome), we obtained information on the early Neolithic funeral practices and on the biogeographical origin of the inhumed individuals. No close kinship was detected. Maternal haplogroups found are consistent with pre-Neolithic settlement, whereas the Y-chromosomal analyses permitted confirmation of the existence in Spain approximately 7,000 y ago of two haplogroups previously associated with the Neolithic transition: G2a and E1b1b1a1b. These results are highly consistent with those previously found in Neolithic individuals from French Late Neolithic individuals, indicating a surprising temporal genetic homogeneity in these groups. The high frequency of G2a in Neolithic samples in western Europe could suggest, furthermore, that the role of men during Neolithic dispersal could be greater than currently estimated.

Published
2011

6. Prognostic value of phosphorylated STAT3 in advanced rectal cancer: a study from 104 French patients included in the EORTC 22921 trial

Author

Bernadette Kantelip, Harouna Zaki, Francine Arbez-Gindre, Mariette Mercier, Franck Monnien, Christophe Borg, Christiane Mougin, Jean-François Bosset, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Service d'oncologie Médicale, Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz, Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Laboratoire d'anatomie pathologique [Besancon], Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz-Université de Franche-Comté ( UFC ), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Service d'Oncologie Médicale [CHRU Besançon], Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Service d'Anatomie pathologique [CHRU Besançon], and Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)

Subjects
Male, Oncology, MESH: Neoplasm Proteins, Pathology, MESH: Combined Modality Therapy, Colorectal cancer, medicine.medical_treatment, MESH : Aged, medicine.disease_cause, MESH: Epidemiologic Methods, law.invention, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH : Phosphorylation, 0302 clinical medicine, MESH: Aged, 80 and over, Randomized controlled trial, law, MESH : Tumor Markers, Biological, MESH : STAT3 Transcription Factor, MESH : Female, Phosphorylation, MESH : Rectal Neoplasms, MESH: Treatment Outcome, Aged, 80 and over, MESH: Aged, 0303 health sciences, MESH : Cell Nucleus, MESH: Middle Aged, MESH : Prognosis, MESH: STAT3 Transcription Factor, Anatomical pathology, MESH : Chemotherapy, Adjuvant, General Medicine, Middle Aged, MESH : Adult, Prognosis, Combined Modality Therapy, Neoplasm Proteins, 3. Good health, Treatment Outcome, Chemotherapy, Adjuvant, MESH: Chemotherapy, Adjuvant, 030220 oncology & carcinogenesis, Female, France, Adult, STAT3 Transcription Factor, MESH: Cell Nucleus, medicine.medical_specialty, MESH : Male, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH : Treatment Outcome, MESH: Prognosis, MESH : Epidemiologic Methods, Pathology and Forensic Medicine, 03 medical and health sciences, MESH : Neoplasm Proteins, Internal medicine, Biomarkers, Tumor, medicine, Humans, MESH : Middle Aged, MESH : Aged, 80 and over, MESH : France, Aged, 030304 developmental biology, Cell Nucleus, Chemotherapy, MESH: Humans, Oncogene, MESH: Phosphorylation, Rectal Neoplasms, business.industry, MESH : Humans, Cancer, MESH: Rectal Neoplasms, MESH: Adult, medicine.disease, MESH: Male, Clinical trial, MESH: France, MESH: Tumor Markers, Biological, Epidemiologic Methods, Carcinogenesis, business, MESH : Combined Modality Therapy, MESH: Female
Abstract

International audience; BACKGROUND: Signal transducer and activator of transcription 3 (STAT3) has been implicated as an oncogene in several neoplastic diseases. However, the biological effects of STAT3 have not been extensively studied in rectal carcinogenesis. AIMS: To evaluate STAT3 activation in advanced rectal cancers and its association with clinicopathological variables and prognosis. METHODS: Nuclear immunohistochemical expression of phosphorylated STAT3 (p-STAT3) was studied in 104 advanced rectal cancers (T3-T4). All patients were participating in the EORTC 22921 trial to assess whether preoperative chemoradiotherapy followed by postoperative chemotherapy improved overall and progression-free survival. RESULTS: Nuclear p-STAT3 expression was detected in 37.5% of rectal cancer patients. No correlation was observed between p-STAT3 and any clinicopathological variables tested. However, patients with tumours positive for p-STAT3 had significantly improved overall survival. CONCLUSION: These results highlight an unexpected role for nuclear p-STAT3 expression in advanced rectal cancers and need further investigation to clarify this finding.

Published
2010

7. 1q12 chromosome translocations form aberrant heterochromatic foci associated with changes in nuclear architecture and gene expression in B cell lymphoma

Author

Stéphanie Chauvelier, Alexandra Debernardi, Katia Delaval, Robert Feil, Remy Gressin, Alexandra Fournier, Martin Figeac, Alicia Lajmanovich, Aurélie Granjon, Christine Lefebvre, Mary Callanan, Yves Usson, Claire Vourc'h, Kassambara Alboukadel, Sophie Rousseaux, John De Vos, Alexei Grichine, Juliana Bruder Ribeyron, Florence de Fraipont, Jean-Pierre Kerckaert, Leila Barki, Thierry Bonnefoix, Anne McLeer-Florin, Saadi Khochbin, Sieme Hamaidia, Samuel Duley, Dominique Leroux, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Département de cancérologie et d'hématologie, CHU Grenoble-Hôpital Michallon, CHU Grenoble, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble, INSERM U823, équipe 5 (cibles diagnostiques ou thérapeutiques et vectorisation de drogues dans le cancer du poumon), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Unité de Cancérologie Biologique et biothérapies, Génomique (Plate-Forme) - Genomics Platform, Institut Pasteur [Paris] (IP), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Institut de recherche en biothérapie (IRB), Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), RFMQ, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 10 (Stress et Dynamique de l'Organisation du Génome), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), INSERM U823, équipe 6 (Epigénétique et Signalisation Cellulaire), INSERM U823, équipe 7 (Voies Oncogéniques Des Hémopathies Malignes), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de cancérologie et d'hématologie, CHU Grenoble-Hôpital Michallon-Hôpital Michallon, Funding was from the Fondation de France, the Institut National du Cancer (EpiPro network, the programme 'Developpement de plateformes hospitalières de génétique moléculaire des cancers', the P.A.I.R 'Mantle Cell Lymphoma network' and the INCa-DHOS translational research programme 'CT-Lymph'). Additional funding was obtained from the Région Rhone-Alpes programme 'Thématique prioritaire--Cancer', the canceropôle CLARA (EpiMed), the Ligue Nationale Contre le Cancer (LNCC)--Comités de l'Isère/Savoie, the Délegation Régionale de la Recherche Clinique--CHU de Grenoble, the French GOELAMS clinical trials group and the ARAMIS association. AD and JBR have been recruited through the INCa-DHOS 'CT-Lymph' program. AF has been the recipient of doctoral funding from the Ministère de l'Enseignement Supérieur et de la Recherche, the Association pour la Recherche sur le Cancer (ARC) and the Société Française d'Hématologie (SFH), AMLF of post-doctoral funding from the ARC-ARECA network 'epigenetic profiling in breast and haematological tumours', LB of doctoral funding from the Ministère de l'Enseignement Supérieur et de la Recherche and the LNCC, MC of a Contrat d'Interface INSERM/Grenoble University Hospital Centre (2006-2011). Affymetrix microarrays were processed in the Microarray Core Facility of the Institute of Research on Biotherapy, CHRU-INSERM-UM1 Montpellier, http://irb.chu-montpellier.fr, Institut d'oncologie/développement Albert Bonniot de Grenoble ( INSERM U823 ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble, Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Unité de Cancérologie Biologique et biothérapies, Génomique (Plate-Forme), Institut Pasteur [Paris], Centre de recherche Jean-Pierre Aubert-Neurosciences et Cancer, Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Lille, Droit et Santé, Institut de recherche en biothérapie ( IRB ), Université Montpellier 1 ( UM1 ) -Centre Hospitalier Régional Universitaire [Montpellier] ( CHRU Montpellier ) -Université de Montpellier ( UM ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications [Grenoble] ( TIMC-IMAG ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Département de cancérologie et d'hématologie, Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille, Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1)-Université de Montpellier (UM), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), and Duley, Samuel

Subjects
Genome instability, Chromosomal translocation, [SDV.GEN] Life Sciences [q-bio]/Genetics, MESH : Chromosomes, Human, Pair 2, MESH : Chromosomes, Human, Pair 1, Translocation, Genetic, 0302 clinical medicine, MESH : Lymphoma, B-Cell, MESH : Translocation, Genetic, Heterochromatin, B-cell lymphoma, 0303 health sciences, MESH : Cell Nucleus, GMCL1, MESH: Gene Expression Regulation, Neoplastic, MESH: Translocation, Genetic, Gene Expression Regulation, Neoplastic, MESH: Heterochromatin, Chromosomes, Human, Pair 1, 030220 oncology & carcinogenesis, Chromosomes, Human, Pair 2, heterochromatic foci, Molecular Medicine, MESH: Cell Nucleus, Lymphoma, B-Cell, MESH : Gene Expression Regulation, Neoplastic, MESH: Chromosomes, Human, Pair 1, MESH: Chromosomes, Human, Pair 2, Biology, 1q12 pericentric heterochromatin, 03 medical and health sciences, medicine, Constitutive heterochromatin, Humans, Epigenetics, 030304 developmental biology, MESH: Lymphoma, B-Cell, Cell Nucleus, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Humans, MESH : Heterochromatin, MESH : Humans, medicine.disease, B cell lymphoma, nuclear architecture, Cancer research, Heterochromatin protein 1, [ SDV.GEN ] Life Sciences [q-bio]/Genetics, Diffuse large B-cell lymphoma, Reports
Abstract

International audience; Epigenetic perturbations are increasingly described in cancer cells where they are thought to contribute to deregulated gene expression and genome instability. Here, we report the first evidence that a distinct category of chromosomal translocations observed in human tumours--those targeting 1q12 satellite DNA--can directly mediate such perturbations by promoting the formation of aberrant heterochromatic foci (aHCF). By detailed investigations of a 1q12 translocation to chromosome 2p, in a case of human B cell lymphoma, aberrant aHCF were shown to be localized to the nuclear periphery and to arise as a consequence of long range 'pairing' between the translocated 1q12 and chromosome 2 centromeric regions. Remarkably, adjacent 2p sequences showed increased levels of repressive histone modifications, including H4K20me3 and H3K9me3, and were bound by HP1. aHCF were associated to aberrant spatial localization and deregulated expression of a novel 2p gene (GMCL1) that was found to have prognostic impact in diffuse large B cell lymphoma. Thus constitutive heterochromatin rearrangements can contribute to tumourigenesis by perturbing gene expression via long range epigenetic mechanisms.

Published
2010

8. Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection

Author

Dolores Vaira, Georges Herbein, Stéphane De Wit, Jean-Stéphane Gatot, Sophie Reuse, Solène Poutrel, Yvan de Launoit, Miriam Calao, Olivier Lambotte, Carine Van Lint, Caroline Vanhulle, Thomas Cherrier, Christine Rouzioux, Jacques Piette, Kabamba Kabeya, Vincent Quivy, Olivier Rohr, Michel Moutschen, Dominique Demonte, Aurélia Lamine, Arsène Burny, Véronique Avettand, Nathan Clumeck, Valérie Martinelli, Yves Collette, Emmanuelle Veithen, Allan Guiguen, Unité de Recherche en Biologie Moléculaire, Facultés Universitaires Notre Dame de la Paix (FUNDP) - Namur, Institut de Biologie de Lille, Institut Pasteur de Lille, Réseau International des Instituts Pasteur ( RIIP ) -Réseau International des Instituts Pasteur ( RIIP ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Virologie [CHU Necker], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Agents pathogènes et inflammation - UFC (EA 4266) ( API ), Université de Franche-Comté ( UFC ), Service de médecine interne et maladies infectieuses, Université Paris-Sud - Paris 11 ( UP11 ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Bicêtre, Division of Infectious Diseases, CHR La réunion, Facultés Universitaires Notre Dame de la Paix (FUNDP), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Centre National de la Recherche Scientifique (CNRS), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Agents pathogènes et inflammation - UFC (EA 4266) (API), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), and Hôpital Bicêtre-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris-Sud - Paris 11 (UP11)

Subjects
Cytoplasm, Anti-HIV Agents -- pharmacology, MESH: I-kappa B Proteins, MESH : Aged, MESH: NF-kappa B, HIV Infections, CD8-Positive T-Lymphocytes, MESH: Monocytes, MESH: Drug Synergism, Monocytes, Phorbol Esters -- pharmacology, Virus latency, MESH: Anti-HIV Agents, Enzyme Inhibitors, lcsh:Science, MESH : Cell Nucleus, 0303 health sciences, education.field_of_study, MESH: Middle Aged, NF-kappa B, Drug Synergism, MESH: HIV Infections, Sciences bio-médicales et agricoles, MESH: CD8-Positive T-Lymphocytes, 3. Good health, Nucleosomes, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Viral load, MESH: Cell Nucleus, MESH : Cytoplasm, MESH : Drug Synergism, HIV Infections -- drug therapy, HIV-1 -- metabolism, Nucleosomes -- metabolism, [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH : Phorbol Esters, 03 medical and health sciences, MESH: Nucleosomes, MESH : HIV Infections, Humans, MESH : Middle Aged, education, Prostratin, Aged, MESH: Humans, 030306 microbiology, MESH : Humans, lcsh:R, MESH: Adult, Molecular Biology/Transcription Initiation and Activation, medicine.disease, chemistry, Viral replication, HIV-1 -- enzymology, HIV-1, lcsh:Q, Histone deacetylase, Cell Nucleus -- metabolism, CD8-Positive T-Lymphocytes -- metabolism, lcsh:Medicine, MESH: HIV-1, chemistry.chemical_compound, NF-KappaB Inhibitor alpha, Phorbol Esters, Enzyme Inhibitors -- pharmacology, MESH : Virus Latency, MESH: Phorbol Esters, MESH: Aged, Multidisciplinary, MESH: Virus Latency, MESH : Adult, MESH : CD8-Positive T-Lymphocytes, Infectious Diseases/HIV Infection and AIDS, Middle Aged, Virus Latency, Cytoplasm -- metabolism, Virology/Viral Replication and Gene Regulation, MESH: Enzyme Inhibitors, MESH : NF-kappa B, I-kappa B Proteins, MESH : HIV-1, Research Article, I-kappa B Proteins -- metabolism, Adult, Monocytes -- virology, Virus Latency -- drug effects, Anti-HIV Agents, Population, MESH : Nucleosomes, Biology, medicine, 030304 developmental biology, Cell Nucleus, MESH : Enzyme Inhibitors, MESH: Cytoplasm, MESH : Anti-HIV Agents, Virology/Persistence and Latency, NF-kappa B -- antagonists & inhibitors, Molecular biology, IκBα, MESH : Monocytes, Cell culture, Cancer research, MESH : I-kappa B Proteins
Abstract

The persistence of transcriptionally silent but replication-competent HIV-1 reservoirs in Highly Active Anti-Retroviral Therapy (HAART)-treated infected individuals, represents a major hurdle to virus eradication. Activation of HIV-1 gene expression in these cells together with an efficient HAART has been proposed as an adjuvant therapy aimed at decreasing the pool of latent viral reservoirs. Using the latently-infected U1 monocytic cell line and latently-infected J-Lat T-cell clones, we here demonstrated a strong synergistic activation of HIV-1 production by clinically used histone deacetylase inhibitors (HDACIs) combined with prostratin, a non-tumor-promoting nuclear factor (NF)- kappaB inducer. In J-Lat cells, we showed that this synergism was due, at least partially, to the synergistic recruitment of unresponsive cells into the expressing cell population. A combination of prostratin+HDACI synergistically activated the 5' Long Terminal Repeat (5'LTR) from HIV-1 Major group subtypes representing the most prevalent viral genetic forms, as shown by transient transfection reporter assays. Mechanistically, HDACIs increased prostratin-induced DNA-binding activity of nuclear NF-kappaB and degradation of cytoplasmic NF-kappaB inhibitor, IkappaBalpha .Moreover, the combined treatment prostratin+HDACI caused a more pronounced nucleosomal remodeling in the U1 viral promoter region than the treatments with the compounds alone. This more pronounced remodeling correlated with a synergistic reactivation of HIV-1 transcription following the combined treatment prostratin+HDACI, as demonstrated by measuring recruitment of RNA polymerase II to the 5'LTR and both initiated and elongated transcripts. The physiological relevance of the prostratin+HDACI synergism was shown in CD8(+)-depleted peripheral blood mononuclear cells from HAART-treated patients with undetectable viral load. Moreover, this combined treatment reactivated viral replication in resting CD4(+) T cells isolated from similar patients. Our results suggest that combinations of different kinds of proviral activators may have important implications for reducing the size of latent HIV-1 reservoirs in HAART-treated patients., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published

Published
2009

9. The histone-binding protein COPR5 is required for nuclear functions of the protein arginine methyltransferase PRMT5

Author

Matthieu Lacroix, Geneviève Rodier, Claude Sardet, Aphonse Le Cam, Eric Fabbrizio, Selma El Messaoudi, Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Génétique Moléculaire de Montpellier (IGMM), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)

Subjects
MESH : Molecular Sequence Data, MESH: Protein Structure, Secondary, MESH: Amino Acid Sequence, MESH : Protein Methyltransferases, Biochemistry, Protein Structure, Secondary, MESH: Recombinant Proteins, Histones, 0302 clinical medicine, MESH: DNA Methylation, Genes, Reporter, MESH : Arginine, Histone methylation, Histone code, Luciferases, Glutathione Transferase, MESH: Histones, 0303 health sciences, MESH : Cell Nucleus, MESH : Chromatin, MESH : Amino Acid Sequence, MESH: Arginine, Nuclear Proteins, MESH : Protein Binding, MESH : Genes, Reporter, Chromatin, Recombinant Proteins, 3. Good health, 030220 oncology & carcinogenesis, Histone methyltransferase, MESH : Carrier Proteins, Protein Binding, MESH: Cell Nucleus, MESH : Glutathione Transferase, MESH: Cell Line, Tumor, MESH : Recombinant Proteins, Molecular Sequence Data, Scientific Report, MESH: Carrier Proteins, Biology, Arginine, MESH: Chromatin, Histone H4, 03 medical and health sciences, Histone H3, Histone H1, Histone arginine methylation, Cell Line, Tumor, Histone H2A, MESH: Protein Methyltransferases, Genetics, MESH: Protein Binding, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Histones, Amino Acid Sequence, Protein Methyltransferases, Molecular Biology, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, Cell Nucleus, MESH : Luciferases, MESH: Glutathione Transferase, MESH: Humans, MESH: Molecular Sequence Data, MESH : Cell Line, Tumor, MESH: Genes, Reporter, MESH : Humans, MESH : Nuclear Proteins, DNA Methylation, MESH: Luciferases, MESH : Protein Structure, Secondary, Carrier Proteins, MESH: Nuclear Proteins, MESH : DNA Methylation
Abstract

International audience; Protein arginine methyltransferase 5 (PRMT5) targets nuclear and cytoplasmic proteins. Here, we identified a nuclear protein, called cooperator of PRMT5 (COPR5), involved in the nuclear functions of PRMT5. COPR5 tightly binds to PRMT5, both in vitro and in living cells, but not to other members of the PRMT family. PRMT5 bound to COPR5 methylates histone H4 (R3) preferentially when compared with histone H3 (R8), suggesting that COPR5 modulates the substrate specificity of nuclear PRMT5-containing complexes, at least towards histones. Markedly, recombinant COPR5 binds to the amino terminus of histone H4 and is required to recruit PRMT5 to reconstituted nucleosomes in vitro. Consistently, COPR5 depletion in cells strongly reduces PRMT5 recruitment on chromatin at the PRMT5 target gene cyclin E1 (CCNE1) in vivo. Moreover, both COPR5 depletion and overexpression affect CCNE1 promoter expression. We propose that COPR5 is an important chromatin adaptor for PRMT5 to function on a subset of its target genes.

Published
2008

10. Impairment of CDKL5 nuclear localisation as a cause for severe infantile encephalopathy

Author

Sylvie Odent, Jamel Chelly, Christophe Philippe, Thierry Bienvenu, Haydeé Rosas-Vargas, L. Lazaro, B. Girard, C. Gitiaux, Nadia Bahi-Buisson, Juliette Nectoux, Philippe Jonveaux, M. A. N'Guyen Morel, Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Unité de génétique médicale, hôpital Sud, Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin ( UMR_S567 / UMR 8104 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Hôpital Sud, Institut de Génétique et Développement de Rennes ( IGDR ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -IFR140-Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-IFR140-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)

Subjects
MESH : RNA, Messenger, MESH: Sequence Homology, Amino Acid, Mutant, CDKL5, MESH: Brain Diseases, Metabolic, Inborn, MESH: Amino Acid Sequence, MESH : Child, Preschool, MESH: Base Sequence, medicine.disease_cause, MESH: Magnetic Resonance Imaging, 0302 clinical medicine, Missense mutation, MESH : Female, MESH: Animals, MESH: DNA Mutational Analysis, [ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics, Genetics (clinical), Genetics, 0303 health sciences, Mutation, MESH : Cell Nucleus, MESH: X Chromosome Inactivation, MESH : Amino Acid Sequence, MESH : Infant, Phenotype, MESH: Amino Acid Substitution, MESH: Infant, Hypotonia, MESH : Sequence Homology, Amino Acid, 3. Good health, MESH: COS Cells, MESH : Phenotype, MESH : X Chromosome Inactivation, medicine.symptom, MESH : Transfection, MESH : COS Cells, MESH : Protein-Serine-Threonine Kinases, MESH: Cell Nucleus, Encephalopathy, MESH : Brain Diseases, Metabolic, Inborn, MESH : DNA Mutational Analysis, Biology, MESH: Phenotype, MESH: Protein-Serine-Threonine Kinases, 03 medical and health sciences, MESH : Amino Acid Substitution, MESH: Plasmids, MESH : Magnetic Resonance Imaging, [ SDV.MHEP ] Life Sciences [q-bio]/Human health and pathology, MESH: Electroencephalography, medicine, Atypical Rett syndrome, MESH : Electroencephalography, 030304 developmental biology, MESH: RNA, Messenger, MESH: Mutation, Missense, MESH: Humans, MESH: Transfection, MESH : Humans, MESH: Child, Preschool, medicine.disease, MESH: Cercopithecus aethiops, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, MESH : Plasmids, MESH : Base Sequence, MESH : Animals, MESH : Cercopithecus aethiops, MESH: Female, 030217 neurology & neurosurgery, MESH : Mutation, Missense, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

International audience; Mutations in the human X-linked cyclin-dependent kinase-like 5 (CDKL5) gene have been shown to cause infantile spasms as well as Rett syndrome-like phenotype. To date, fewer than 20 different mutations have been reported. So far, no clear genotype-phenotype correlation has been established. We screened the entire coding region of CDKL5 in 151 affected girls with a clinically heterogeneous phenotype ranging from encephalopathy with epilepsy to atypical Rett syndrome by denaturing high liquid performance chromatography and direct sequencing, and we identified three novel missense mutations located in catalytic domain (p.Ala40Val, p.Arg65Gln, p.Leu220Pro). Segregation analysis showed that p.Arg65Gln was inherited from the healthy father, which rules out the involvement of CDKL5 in the aetiology of the phenotype in this patient. However, the de novo occurrence was shown for p.Ala40Val and p.Leu220Pro. The p.Ala40Val mutation was observed in two unrelated patients and represented the first recurrent mutation in the CDKL5 gene. For the two de novo mutations, we analysed the cellular localisation of the wild-type and CDKL5 mutants by transfection experiments. We showed that the two CDKL5 mutations cause mislocalisation of the mutant CDKL5 proteins in the cytoplasm. Interestingly these missense mutations that result in a mislocalisation of the CDKL5 protein are associated with severe developmental delay which was apparent within the first months of life characterised by early and generalised hypotonia, and autistic features, and as well as early infantile spasms.

Published
2008

11. Cell-cell fusion induced by measles virus amplifies the type I interferon response

Author

Johan Druelle, Timothy Fabian Wild, Florence Herschke, Denis Gerlier, Sébastien Plumet, Chantal Rabourdin-Combe, David Laine, Hélène Valentin, Olga Azocar, Thomas Duhen, Virologie et Pathologie Humaine (VirPath), 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), Immunobiologie fondamentale et clinique, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), BioSciences Lyon-Gerland (BLG), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Virologie humaine, École normale supérieure - Lyon (ENS Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported in part by grants from ANR (DG, ANR-MIME), INSERM, INCA-Canceropole 2004-2005 and ARC 3637. D.L., F.H., J.D., T.D., and S.P. were supported by a fellowship from MENRT, ARC and DGA, respectively., Measles and Interferon response, École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), École normale supérieure de Lyon (ENS de Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), Virologie et Pathologie Humaine ( VirPath ), 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 ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), BioSciences Lyon-Gerland ( BLG ), École normale supérieure - Lyon ( ENS Lyon ) -Institut National de la Recherche Agronomique ( INRA ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Hospices Civils de Lyon ( HCL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), École normale supérieure - Lyon ( ENS Lyon ) -IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Gerlier, Denis

Subjects
MESH : Cell Line, MESH: Interferon Type I, fusion, Interferon Regulatory Factor-7, MESH : Viral Fusion Proteins, Cellular Response to Infection, Giant Cells, MESH: Interferon Regulatory Factor-7, MESH : Virology, Cell Fusion, MESH: Giant Cells, MESH : Interferon Regulatory Factor-7, MESH: Interferon Regulatory Factor-3, Interferon, Chlorocebus aethiops, MESH : Interferon Regulatory Factor-3, MESH : Innate immunity, MESH: Animals, MESH : Viral Proteins, Mononegavirales, OCIS 000.1430, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH : Cell Nucleus, Cell fusion, Microscopy, Video, biology, MESH: Dendritic Cells, MESH : Microscopy, Video, interferon, MESH: Innate immunity, MESH : Epithelial Cells, MESH: Epithelial Cells, Interferon Type I, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Viral Fusion Proteins, medicine.drug, MESH: Cell Nucleus, Immunology, Microbiology, [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Virus, Cell Line, Measles virus, Viral Proteins, MESH : Interferon Type I, Virology, medicine, Animals, Humans, dendritic cells, MESH: Immunology, Cell Nucleus, MESH: Humans, MESH : Cell Fusion, MESH : Humans, MESH : Giant Cells, biology.organism_classification, Molecular biology, MESH: Viral Proteins, MESH: Cercopithecus aethiops, epithelial cells, MESH: Cell Line, MESH: Microscopy, Video, Giant cell, Cell culture, Insect Science, MESH : Dendritic Cells, measles virus, MESH: Cell Fusion, MESH : Immunology, MESH : Measles virus, Interferon Regulatory Factor-3, MESH : Animals, MESH : Cercopithecus aethiops, Viral Fusion Proteins, MESH: Measles virus, Interferon type I, MESH: Virology
Abstract

Measles virus (MeV) infection is characterized by the formation of multinuclear giant cells (MGC). We report that beta interferon (IFN-β) production is amplified in vitro by the formation of virus-induced MGC derived from human epithelial cells or mature conventional dendritic cells. Both fusion and IFN-β response amplification were inhibited in a dose-dependent way by a fusion-inhibitory peptide after MeV infection of epithelial cells. This effect was observed at both low and high multiplicities of infection. While in the absence of virus replication, the cell-cell fusion mediated by MeV H/F glycoproteins did not activate any IFN-α/β production, an amplified IFN-β response was observed when H/F-induced MGC were infected with a nonfusogenic recombinant chimerical virus. Time lapse microscopy studies revealed that MeV-infected MGC from epithelial cells have a highly dynamic behavior and an unexpected long life span. Following cell-cell fusion, both of the RIG-I and IFN-β gene deficiencies were trans complemented to induce IFN-β production. Production of IFN-β and IFN-α was also observed in MeV-infected immature dendritic cells (iDC) and mature dendritic cells (mDC). In contrast to iDC, MeV infection of mDC induced MGC, which produced enhanced amounts of IFN-α/β. The amplification of IFN-β production was associated with a sustained nuclear localization of IFN regulatory factor 3 (IRF-3) in MeV-induced MGC derived from both epithelial cells and mDC, while the IRF-7 up-regulation was poorly sensitive to the fusion process. Therefore, MeV-induced cell-cell fusion amplifies IFN-α/β production in infected cells, and this indicates that MGC contribute to the antiviral immune response.

Published
2007

12. Bsr, a nuclear-retained RNA with monoallelic expression

Author

Jérôme Cavaillé, Maud Marques, Eugenia Basyuk, Edouard Bertrand, Virginie Marty, Hélène Royo, Laboratoire de biologie moléculaire eucaryote du CNRS ( LBME ), Université Paul Sabatier - Toulouse 3 ( UPS ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Génomique Fonctionnelle ( IGF ), Centre National de la Recherche Scientifique ( CNRS ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Montpellier 1 ( UM1 ) -Université de Montpellier ( UM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Biologie du Développement de Marseille ( IBDM ), Aix Marseille Université ( AMU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de Recherches sur la Cognition Animale [Toulouse] ( CRCA ), 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 ), Departament Geodinàmica i Geofísica, Facultat de Geologia, Laboratoire de Physique Statistique de l'ENS ( LPS ), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris ( FRDPENS ), Centre National de la Recherche Scientifique ( CNRS ) -École normale supérieure - Paris ( ENS Paris ) -Centre National de la Recherche Scientifique ( CNRS ) -École normale supérieure - Paris ( ENS Paris ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire Colloïdes et Matériaux Divisés ( LCMD ), ESPCI ParisTech-Centre National de la Recherche Scientifique ( CNRS ), Liquides Ioniques et Interfaces Chargées ( LI2C ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -ESPCI ParisTech-Centre National de la Recherche Scientifique ( CNRS ), Ecole Nationale de la Santé Publique ( ENSP ), École des Hautes Études en Santé Publique [EHESP] ( EHESP ), Centre de recherches Paul Pascal ( CRPP ), Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de biologie moléculaire eucaryote (LBME), Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut de Biologie du Développement de Marseille (IBDM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherches sur la Cognition Animale (CRCA), Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut des sciences du cerveau de Toulouse. (ISCT), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse - Jean Jaurès (UT2J)-CHU Toulouse [Toulouse]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Département de biologie [Sherbrooke] (UdeS), Faculté des sciences [Sherbrooke] (UdeS), Université de Sherbrooke (UdeS)-Université de Sherbrooke (UdeS), Laboratoire de Physique Statistique de l'ENS (LPS), Université Paris Diderot - Paris 7 (UPD7)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Colloïdes et Matériaux Divisés (LCMD), Centre National de la Recherche Scientifique (CNRS)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Liquides Ioniques et Interfaces Chargées (LI2C), Université Pierre et Marie Curie - Paris 6 (UPMC)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Ecole Nationale de la Santé Publique (ENSP), École des Hautes Études en Santé Publique [EHESP] (EHESP), Centre de recherches Paul Pascal (CRPP), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherches sur la Cognition Animale - UMR5169 (CRCA), Institut des sciences du cerveau de Toulouse. (ISCT), Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-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 - Jean Jaurès (UT2J), Université de Toulouse (UT)-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)-Centre de Biologie Intégrative (CBI), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), and Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris)

Subjects
MESH: Hydroxamic Acids, RNA, Untranslated, MESH : RNA, Messenger, MESH: Introns, RNA Stability, Hydroxamic Acids, RNA Transport, MESH: RNA, Untranslated, 0302 clinical medicine, MESH: RNA Transport, Transcription (biology), Gene cluster, MESH : RNA, MESH: Animals, MESH : Cell Nucleus Structures, MESH : Female, MESH : RNA Transport, Genetics, 0303 health sciences, MESH : Cell Nucleus, MESH : Rats, MESH: RNA Stability, Articles, MESH : RNA Stability, Non-coding RNA, medicine.anatomical_structure, RNA splicing, Female, MESH : RNA Splicing, MESH: Cell Nucleus, MESH : RNA, Untranslated, MESH: Rats, RNA Splicing, MESH: Cytoplasmic Granules, Biology, Cytoplasmic Granules, MESH : Rats, Wistar, MESH : Introns, 03 medical and health sciences, MESH: RNA, MESH : Fibroblasts, medicine, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Cell Nucleus Structures, RNA, Messenger, Rats, Wistar, Molecular Biology, Gene, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Alleles, MESH: RNA, Messenger, 030304 developmental biology, Cell Nucleus, MESH : Hydroxamic Acids, MESH: Alleles, MESH : Cytoplasmic Granules, Intron, RNA, MESH: Rats, Wistar, Cell Biology, Fibroblasts, Cell Nucleus Structures, Introns, Rats, Cell nucleus, MESH: Fibroblasts, MESH : Animals, MESH: RNA Splicing, MESH : Alleles, MESH: Female, 030217 neurology & neurosurgery
Abstract

The imprinted Dlk1-Gtl2 and Prader-Willi syndrome (PWS) regions are characterized by a complex noncoding transcription unit spanning arrays of tandemly repeated C/D RNA genes. These noncoding RNAs (ncRNAs) are thought to play an essential but still poorly understood role. To better understand the intracellular fate of these large ncRNAs, fluorescence in situ hybridization was carried out at the rat Dlk1-Gtl2 domain. This locus contains a ∼100-kb-long gene cluster comprising 86 homologous RBII-36 C/D RNA gene copies, all of them intron-encoded within the ncRNA gene Bsr. Here, we demonstrate that the Bsr gene is monoallelically expressed in primary rat embryonic fibroblasts as well as in hypothalamic neurons and yields a large amount of unspliced and spliced RNAs at the transcription site, mostly as elongated RNA signals. Surprisingly, spliced Bsr RNAs released from the transcription site mainly concentrate as numerous, stable nuclear foci that do not colocalize with any known subnuclear structures. On drug treatments, a fraction of Bsr RNA relocalizes to the cytoplasm and associates with stress granules (SGs), but not with P-bodies, pointing to a potential link between SGs and the metabolism of ncRNA. Thus, Bsr might represent a novel type of nuclear-retained transcript.

Published
2007

13. Inflammation of choriodecidua induces tumor necrosis factor alpha-mediated apoptosis of human myometrial cells

Author

Leroy , Marie-Josèphe, Dallot , Emmanuelle, Czerkiewicz , Isabelle, Schmitz , Thomas, Breuiller-Fouché , Michelle, Breuiller-Fouche, Michelle, La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus (UMR_S 767), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Maternité Port-Royal [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus ( UMR_S 767 ), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP]

Subjects
Lipopolysaccharides, Placenta, Extraembryonic Membranes, Apoptosis, MESH : Blotting, Western, Receptors, Tumor Necrosis Factor, MESH: Decidua, Pregnancy, MESH : Female, MESH : Inflamma, MESH: In Situ Nick-End Labeling, MESH : Culture Media, Conditioned, Cells, Cultured, MESH : Cell Nucleus, MESH : Cell Survival, MESH: Culture Media, Conditioned, MESH: Enzyme-Linked Immunosorbent Assay, MESH : Enzyme-Linked Immunosorbent Assay, MESH : Extraembryonic Membranes, MESH: Cell Survival, MESH : In Situ Nick-End Labeling, Myometrium, MESH : Amnion, Female, MESH: Cells, Cultured, MESH: Cell Nucleus, MESH : Decidua, Cell Survival, Blotting, Western, Enzyme-Linked Immunosorbent Assay, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Cells, Cultured, MESH: Antibodies, Blocking, Decidua, In Situ Nick-End Labeling, Humans, MESH: Blotting, Western, Amnion, Antibodies, Blocking, MESH: Amnion, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Cell Nucleus, Inflammation, MESH: Inflamma, MESH : Antibodies, Blocking, MESH: Humans, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, Tumor Necrosis Factor-alpha, MESH: Extraembryonic Membranes, MESH: Apoptosis, MESH : Humans, Toll-Like Receptor 4, Culture Media, Conditioned, MESH: Female, MESH : Apoptosis
Abstract

The present study investigated the ability of human choriodecidua to induce myometrial cell apoptosis through the secretion of tumor necrosis factor alpha (TNF). The secretion of TNF was evaluated in the culture supernatants of amnion and choriodecidua explants that were exposed to the bacterial endotoxin lipopolysaccharide (LPS) to mimic inflammation. The choriodecidua explants produced more TNF than the amnion explants in response to LPS stimulation, despite the fact that the choriodecidua had lower levels of TLR4 expression. Moreover, conditioned medium obtained from LPS-treated choriodecidua explants, but not that from amnion explants, decreased the number of viable cultured myometrial cells and induced cell apoptosis by inducing the overexpression of the proapoptotic protein BAX and by decreasing the expression of the anti-apoptotic protein BCL2. Neutralization of TNF in the choriodecidua-conditioned medium reversed this effect. Exogenous TNF mimicked LPS-treated choriodecidua-conditioned medium in that it induced myometrial cell apoptosis, reduced BCL2 expression, and increased BAX expression. Using neutralizing antibodies against both subtypes of TNF receptors, we found that only TNFRSF1A participates in TNF-induced myometrial cell apoptosis. Our in vitro model of LPS-induced inflammation of human fetal membrane explants suggests a mechanism by which TNF secreted by choriodecidua governs human myometrial cell apoptosis at the end of pregnancy. These data support the hypothesis that TNF participates in the complex network of signaling processes associated with uterine involution.

Published
2007

14. Purification techniques of subcellular compartments for analytical and preparative purposes

Author

Aubry , Laurence, Klein , Gérard, Biochimie et biophysique des systèmes intégrés (BBSI), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Biochimie et biophysique des systèmes intégrés ( BBSI ), and Université Joseph Fourier - Grenoble 1 ( UJF ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
MESH: Cell Nucleus, MESH: Mitochondria, MESH : Cell Membrane, MESH: Dictyostelium, Endosomes, MESH : Subcellular Fractions, Cell Fractionation, MESH : Dictyostelium, MESH : Organelles, Animals, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Dictyostelium, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Cell Nucleus, Organelles, MESH : Cell Nucleus, Cell Membrane, MESH : Lysosomes, Mitochondria, MESH: Endosomes, MESH: Subcellular Fractions, MESH : Cell Fractionation, MESH : Endosomes, MESH: Cell Fractionation, MESH : Animals, MESH : Mitochondria, Lysosomes, MESH: Cell Membrane, MESH: Lysosomes, MESH: Organelles, Subcellular Fractions
Abstract

International audience; In the following protocols, broken cells are the starting material of all downstream purifications of functional organelles or intact subcellular membranes. The choice of the breakage method has direct and deep repercussions on the quality of subsequent steps. Breaking vegetative amoebae by shear stress with a steel ball cell cracker preserves the integrity of subcellular organelles and in particular that of lysosomes, the rupture of which is very deleterious to further purifications. In this chapter, we propose purification schemes for plasma membrane, nuclei, mitochondria, and endocytic compartments. Plasma membranes are purified without any cell coating by partition between aqueous polymer phases. Nuclei and mitochondria are purified by differential centrifugations in adequate buffer conditions. Endosomes are magnetically isolated after feeding the cells with colloidal iron dextran and phagosomes by flotation on a sucrose gradient after feeding amoebae with latex beads. As analytical approaches, we propose procedures to label the plasma membrane and the endo-lysosomal compartments by biotinylation and to separate early and late compartments on a Percoll gradient.

Published
2006

15. Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species

Author

Geert-Jan van Gemert, Jean-François Franetich, Laurent Hannoun, Anne Dubart-Kupperschmitt, Shoshana Levy, Olivier Silvie, Eric Rubinstein, Claude Boucheix, Dominique Mazier, Céline Greco, Robert W. Sauerwein, Immunobiologie Cellulaire et Moléculaire des Infections Parasitaires, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR113-Institut National de la Santé et de la Recherche Médicale (INSERM), Microenvironnement et Physiopathologie de la Differenciation, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Department of Medical Microbiology, Radboud University Medical Center [Nijmegen], Division of Oncology, Department of Medicine, Stanford University Medical Center, Service de Chirurgie Digestive, Hépato-Bilio-pancréatique et Transplantation Hépatique [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Garcia, Marie, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -IFR113-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Cochin ( UMR_S567 / UMR 8104 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Service de chirurgie digestive et hépato-bilio-pancréatique [CHU Pitié-Salpétrière], and Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP]

Subjects
MESH : Sporozoites, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH : Animals, Genetically Modified, Host tropism, MESH: Sporozoites, MESH: Host-Parasite Relations, Plasmodium, Animals, Genetically Modified, Mice, Tetraspanin, MESH : Plasmodium falciparum, MESH: Animals, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, MESH: Antigens, CD, MESH: Plasmodium falciparum, MESH: Plasmodium yoelii, MESH : Cell Nucleus, 0303 health sciences, biology, 3. Good health, Sporozoites, Infection and autoimmunity [NCMLS 1], [ INFO.INFO-BT ] Computer Science [cs]/Biotechnology, Plasmodium yoelii, MESH: Cell Nucleus, MESH: Cell Line, Tumor, Plasmodium falciparum, Immunology, Auto-immunity, transplantation and immunotherapy [N4i 4], Microbiology, Host-Parasite Interactions, Tetraspanin 28, MESH: Animals, Genetically Modified, 03 medical and health sciences, Species Specificity, Antigen, Antigens, CD, Cell Line, Tumor, Virology, MESH : Mice, parasitic diseases, MESH : Antigens, CD, Animals, Humans, MESH : Species Specificity, MESH: Species Specificity, MESH: Mice, 030304 developmental biology, Cell Nucleus, MESH: Humans, MESH : Cell Line, Tumor, 030306 microbiology, MESH : Humans, fungi, Poverty-related infectious diseases [N4i 3], [ SDV.BIO ] Life Sciences [q-bio]/Biotechnology, MESH : Plasmodium yoelii, MESH : Host-Parasite Relations, biology.organism_classification, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, [INFO.INFO-BT] Computer Science [cs]/Biotechnology, Cell culture, MESH : Animals, Microbial pathogenesis and host defense [UMCN 4.1], Immunity, infection and tissue repair [NCMLS 1], CD81
Abstract

Contains fulltext : 49897.pdf (Publisher’s version ) (Closed access) Plasmodium sporozoites can enter host cells by two distinct pathways, either through disruption of the plasma membrane followed by parasite transmigration through cells, or by formation of a parasitophorous vacuole (PV) where the parasite further differentiates into a replicative exo-erythrocytic form (EEF). We now provide evidence that following invasion without PV formation, transmigrating Plasmodium falciparum and Plasmodium yoelii sporozoites can partially develop into EEFs inside hepatocarcinoma cell nuclei. We also found that rodent P. yoelii sporozoites can infect both mouse and human hepatocytes, while human P. falciparum sporozoites infect human but not mouse hepatocytes. We have previously reported that the host tetraspanin CD81 is required for PV formation by P. falciparum and P. yoelii sporozoites. Here we show that expression of human CD81 in CD81-knockout mouse hepatocytes is sufficient to confer susceptibility to P. yoelii but not P. falciparum sporozoite infection, showing that the narrow P. falciparum host tropism does not rely on CD81 only. Also, expression of CD81 in a human hepatocarcinoma cell line is sufficient to promote the formation of a PV by P. yoelii but not P. falciparum sporozoites. These results highlight critical differences between P. yoelii and P. falciparum sporozoite infection, and suggest that in addition to CD81, other molecules are specifically required for PV formation during infection by the human malaria parasite.

Published
2006

16. 7-Ketocholesterol favors lipid accumulation and colocalizes with Nile Red positive cytoplasmic structures formed during 7-ketocholesterol-induced apoptosis: analysis by flow cytometry, FRET biphoton spectral imaging microscopy, and subcellular fractionation

Author

Ginette Bessède, Gérard Lizard, Edmond Kahn, Anne Vejux, Jean-Marc Riedinger, Andrew Todd-Pokropek, Anne Athias, Franck Ménétrier, Frédérique Frouin, Eric Ogier-Denis, Jean-François Stoltz, Dominique Dumas, Laboratoire de Biochimie Moléculaire et Cellulaire ( LBMC ), Université de Bourgogne ( UB ), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Laboratoire d'Imagerie Fonctionnelle ( LIF ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -IFR14-IFR49-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Laboratoire d'Energétique et de Mécanique Théorique Appliquée ( LEMTA ), Université de Lorraine ( UL ) -Centre National de la Recherche Scientifique ( CNRS ), Plate-forme Imagerie et Biophysique Cellulaire et Tissulaire ( PTIBC-IBISA Nancy ), Université Henri Poincaré - Nancy 1 ( UHP ), Plateforme Lipidomique [Dijon] ( LAP ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ) -Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ) -IFR100 - Structure fédérative de recherche Santé-STIC-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Laboratoire de Biologie Médicale [Centre Georges-François leclerc], Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ), Faculté de Médecine Xavier Bichat, Laboratoire de Biochimie Moléculaire et Cellulaire (LBMC), Université de Bourgogne (UB), Lipides - Nutrition - Cancer (U866) (LNC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bourgogne (UB)-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Laboratoire d'Imagerie Fonctionnelle (LIF), Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR14-IFR49-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Énergies et Mécanique Théorique et Appliquée (LEMTA ), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Plate-forme Imagerie et Biophysique Cellulaire et Tissulaire (PTIBC-IBISA Nancy), Université Henri Poincaré - Nancy 1 (UHP), Plateforme Lipidomique [Dijon] (LAP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bourgogne (UB)-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bourgogne (UB)-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-IFR100 - Structure fédérative de recherche Santé-STIC-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER, Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), and Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-IFR100 - Structure fédérative de recherche Santé-STIC-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)

Subjects
MESH: Flow Cytometry, Apoptosis, 7. Clean energy, law.invention, MESH : Phosphatidylcholines, 0302 clinical medicine, Fluorescence Resonance Energy Transfer, MESH : Fluorescent Dyes, Enzyme Inhibitors, MESH: Lipid Metabolism, 0303 health sciences, MESH : Cell Nucleus, Flow Cytometry, MESH: Image Processing, Computer-Assisted, Cholesterol, MESH : Propidium, 030220 oncology & carcinogenesis, MESH : U937 Cells, MESH: Microscopy, Electron, Transmission, MESH: Cell Fractionation, MESH : Image Processing, Computer-Assisted, MESH: Propidium, Propidium, MESH: Cell Nucleus, MESH : Sphingomyelins, MESH: Mitochondria, Cell Fractionation, Filipin, Permeability, Pathology and Forensic Medicine, 03 medical and health sciences, Microscopy, Electron, Transmission, Oxazines, MESH: Membrane Potentials, Humans, Propidium iodide, MESH: Humans, MESH : Oxazines, MESH : Humans, Nile red, MESH: Phosphatidylcholines, Lipid Metabolism, Förster resonance energy transfer, chemistry, MESH: Subcellular Fractions, Cytoplasmic Structures, Benzimidazoles, MESH : Apoptosis, MESH: Fluorescence Resonance Energy Transfer, MESH: Sphingomyelins, Membrane Potentials, chemistry.chemical_compound, MESH: Cholesterol, [ SDV.BBM.BC ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], law, MESH : Lipid Metabolism, Image Processing, Computer-Assisted, Fluorescence microscope, MESH : Membrane Potentials, Ketocholesterols, MESH : Cholesterol, MESH: Ketocholesterols, U937 Cells, MESH: Fluorescent Dyes, Mitochondria, Sphingomyelins, MESH: Intracellular Membranes, MESH : Microscopy, Electron, Transmission, Biochemistry, MESH : Benzimidazoles, MESH: Enzyme Inhibitors, MESH: Permeability, MESH : Cell Fractionation, Phosphatidylcholines, lipids (amino acids, peptides, and proteins), MESH : Mitochondria, Cell fractionation, Subcellular Fractions, Histology, MESH : Flow Cytometry, MESH: U937 Cells, MESH : Subcellular Fractions, Confocal microscopy, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Fluorescent Dyes, 030304 developmental biology, MESH : Ketocholesterols, Cell Nucleus, MESH : Enzyme Inhibitors, MESH: Apoptosis, Lipid metabolism, Intracellular Membranes, Cell Biology, MESH : Cytoplasmic Structures, MESH: Cytoplasmic Structures, MESH : Intracellular Membranes, MESH : Permeability, MESH : Fluorescence Resonance Energy Transfer, MESH: Benzimidazoles, MESH: Oxazines
Abstract

Background Oxidized low-density lipoproteins play key roles in atherosclerosis. Their toxicity is at least in part due to 7-ketocholesterol (7KC), which is a potent inducer of apoptosis. In this study on human promonocytic U937 cells, we determined the effects and the interactions of 7KC with cellular lipids during 7KC-induced apoptosis. Methods Morphologic and functional changes were investigated by microscopic and flow cytometric methods after staining with propidium iodide, 3,3′-dihexyloxacarbocyanine iodide, and Hoechst 33342. Cellular lipid content was identified by using filipin to quantify free cholesterol and Nile Red (NR), which emit a yellow or orange-red fluorescence in the presence of neutral and polar lipids, respectively. After staining with NR, interactions of 7KC with cellular lipids were identified by fluorescence resonance energy transfer biphoton spectral imaging confocal microscopy and by subcellular fractionation, gas chromatography, and mass spectrometry. Results During 7KC-induced apoptosis the fluorescence from filipin and the ratio of measured (orange-red vs. yellow) fluorescence of NR were enhanced. Spectral analysis of images obtained in biphoton mode and resulting factor images demonstrated the occurrence of fluorescence resonance energy transfer between 7KC and NR and the subsequent colocalization of 7KC and NR. These data were in agreement with biochemical characterization and demonstrated that 7KC and neutral and polar lipids accumulate in NR-stained cytoplasmic structures. Conclusions During 7KC-induced apoptosis, 7KC modifies the cellular content of neutral and polar lipids, favors free cholesterol accumulation, and colocalizes with neutral and polar lipids that are inside NR-stained cytoplasmic structures. © 2005 Wiley-Liss, Inc.

Published
2005

17. Nucleocytoplasmic shuttling of antigen in mammalian cells conferred by a soluble versus insoluble single-chain antibody fragment equipped with import/export signals

Author

Pierre Martineau, Etienne Weiss, Annie-Paule Sibler, Alexandra Nordhammer, Murielle Masson, Gilles Travé, Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique ( CNRS ), Material Physics Laboratory, Helsinki University of Technology ( TKK ), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 ( UGSF ), Institut National de la Recherche Agronomique ( INRA ) -Université de Lille-Centre National de la Recherche Scientifique ( CNRS ), Franche-Comté Électronique Mécanique, Thermique et Optique - Sciences et Technologies ( FEMTO-ST ), Université de Technologie de Belfort-Montbeliard ( UTBM ) -Ecole Nationale Supérieure de Mécanique et des Microtechniques ( ENSMM ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Heuristique et Diagnostic des Systèmes Complexes [Compiègne] ( Heudiasyc ), Centre National de la Recherche Scientifique ( CNRS ), Laboratoire d'ingénierie osteo-articulaire et dentaire ( LIOAD ), Université de Nantes ( UN ) -IFR26-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de Recherche en Infectiologie de Montpellier ( IRIM ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Montpellier ( UM ), Laboratoire d'Informatique, de Modélisation et d'optimisation des Systèmes ( LIMOS ), Centre National de la Recherche Scientifique ( CNRS ) -Sigma CLERMONT ( Sigma CLERMONT ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Université Blaise Pascal - Clermont-Ferrand 2 ( UBP ), Université Montpellier 1 ( UM1 ), Laboratoire de génie chimique ( LGC ), Institut National Polytechnique [Toulouse] ( INP ) -Université Paul Sabatier - Toulouse 3 ( UPS ) -Centre National de la Recherche Scientifique ( CNRS ), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), TKK Helsinki University of Technology (TKK), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Franche-Comté Électronique Mécanique, Thermique et Optique - Sciences et Technologies (UMR 6174) (FEMTO-ST), Université de Technologie de Belfort-Montbeliard (UTBM)-Ecole Nationale Supérieure de Mécanique et des Microtechniques (ENSMM)-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Heuristique et Diagnostic des Systèmes Complexes [Compiègne] (Heudiasyc), Université de Technologie de Compiègne (UTC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'ingénierie osteo-articulaire et dentaire (LIOAD), Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Informatique, de Modélisation et d'optimisation des Systèmes (LIMOS), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Sigma CLERMONT (Sigma CLERMONT)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure des Mines de St Etienne, Université Montpellier 1 (UM1), Laboratoire de génie chimique [ancien site de Basso-Cambo] (LGC), 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 Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Biotechnologie des interactions macromoléculaires (BIM), Centre National de la Recherche Scientifique (CNRS), Institut de Biotechnologie-Pharmacologie, Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-BioRad-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), and Université de Franche-Comté (UFC)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Mécanique et des Microtechniques (ENSMM)-Université de Technologie de Belfort-Montbeliard (UTBM)

Subjects
MESH: Signal Transduction, Cytoplasm, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH : Hela Cells, Antigens, Polyomavirus Transforming, [SDV]Life Sciences [q-bio], MESH : Antigens, MESH: Eukaryotic Cells, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, 0302 clinical medicine, MESH : Protein Transport, MESH : Mammals, MESH : Bacterial Proteins, MESH: Animals, Expression in mammalian cells, MESH: Peptide Fragments, MESH: Bacterial Proteins, ComputingMilieux_MISCELLANEOUS, Mammals, 0303 health sciences, Single-chain Fv, MESH : Cell Nucleus, MESH : Protein Binding, Transfection, MESH: COS Cells, Protein Transport, Eukaryotic Cells, Biochemistry, 030220 oncology & carcinogenesis, COS Cells, MESH: Antigens, MESH : COS Cells, Intracellular, Protein Binding, Signal Transduction, MESH: Cell Nucleus, MESH: Protein Transport, MESH : Cytoplasm, Nucleocytoplasmic indirect transport, Intracellular targeting, Active Transport, Cell Nucleus, NLS/NES, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH: Active Transport, Cell Nucleus, Biology, MESH: Mammals, Protein–protein interaction, 03 medical and health sciences, Bacterial Proteins, Antigen, MESH : Prokaryotic Cells, Animals, Humans, NLS, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Antigens, Nuclear export signal, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Active Transport, Cell Nucleus, 030304 developmental biology, MESH: Prokaryotic Cells, Cell Nucleus, MESH : Signal Transduction, MESH: Humans, MESH: Antigens, Polyomavirus Transforming, MESH: Cytoplasm, MESH : Solubility, MESH : Humans, MESH : Peptide Fragments, MESH : Eukaryotic Cells, [ SDV.BIO ] Life Sciences [q-bio]/Biotechnology, Cell Biology, Peptide Fragments, MESH: Hela Cells, MESH: Solubility, Prokaryotic Cells, Solubility, Functional solubility, MESH : Animals, Nuclear transport, MESH : Antigens, Polyomavirus Transforming, HeLa Cells
Abstract

International audience; The ectopic expression of antibody fragments within mammalian cells is a challenging approach for interfering with or even blocking the biological function of the intracellular target. For this purpose, single-chain Fv (scFv) fragments are generally preferred. Here, by transfecting several mammalian cell lines, we compared the intracellular behavior of two scFvs (13R4 and 1F4) that strongly differ in their requirement of disulphide bonding for the formation of active molecules in bacteria. The scFv 13R4, which is correctly folded in the bacterial cytoplasm, was solubly expressed in all cell lines tested and was distributed in their cytoplasm and nucleus, as well. In addition, by appending to the 13R4 molecules the SV40 T-antigen nuclear localisation signal (NLS) tag, cytoplasmic-coexpressed antigen was efficiently retargeted to the nucleus. Compared to the scFv 13R4, the scFv 1F4, which needs to be secreted in bacteria for activity, accumulated, even with the NLS tag, as insoluble aggregates within the cytoplasm of the transfected cells, thereby severely disturbing fundamental functions of cell physiology. Furthermore, by replacing the NLS tag with a leucine-rich nuclear export signal (NES), the scFv 13R4 was exclusively located in the cytoplasm, whereas the similarly modified scFv 1F4 still promoted cell death. Coexpression of NES-tagged 13R4 fragments with nuclear antigen promoted its efficient retargeting to the cytoplasm. This dominant effect of the NES tag was also observed after exchange of the nuclear signals between the scFv 13R4 and its antigen. Taken together, the results indicate that scFvs that are active in the cytoplasm of bacteria may behave similarly in mammalian cells and that the requirement of their conserved disulphide bridges for activity is a limiting factor for mediating the nuclear import/export of target in a mammalian cell context. The described shuttling effect of antigen conferred by a soluble scFv may represent the basis of a reliable in vivo assay of effective protein- protein interactions.

Published
2003

18. Insulin effects on sterol regulatory-element-binding protein-1c (SREBP-1c) transcriptional activity in rat hepatocytes

Author

Dalila Azzout-Marniche, Marc Foretz, Fabienne Foufelle, Pascal Ferré, Colette Guichard, Dominique Bécard, Physiologie de la Nutrition et du Comportement Alimentaire (PNCA), AgroParisTech-Institut National de la Recherche Agronomique (INRA), [Institut Cochin] Département Endocrinologie, métabolisme, diabète (EMD) (EMD), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-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), Centre de Recherche des Cordeliers (CRC), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Pierre et Marie Curie - Paris 6 (UPMC)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiologie de la Nutrition et du Comportement Alimentaire ( PNCA ), Institut National de la Recherche Agronomique ( INRA ) -AgroParisTech, [Institut Cochin] Département Endocrinologie, métabolisme, diabète (EMD) ( EMD ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -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 ), Centre de Recherche des Cordeliers ( CRC ), Université Paris Diderot - Paris 7 ( UPD7 ) -École pratique des hautes études ( EPHE ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Université Pierre et Marie Curie - Paris 6 (UPMC)-École Pratique des Hautes Études (EPHE)

Subjects
MESH: Protein Precursors, Transcription, Genetic, medicine.medical_treatment, MESH : Hepatocytes, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Biochemistry, MESH: CCAAT-Enhancer-Binding Proteins, MESH: Hepatocytes, 0302 clinical medicine, Insulin receptor substrate, Insulin, MESH: Animals, Enzyme Inhibitors, MESH : Immunoblotting, Cells, Cultured, Protein Synthesis Inhibitors, 0303 health sciences, MESH : Cell Nucleus, MESH : Glucokinase, MESH: Immunoblotting, food and beverages, [SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN], MESH: Glucokinase, MESH: Transcription Factors, [ SDV.MHEP.EM ] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, 3. Good health, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 030220 oncology & carcinogenesis, MESH : DNA-Binding Proteins, Sterol Regulatory Element Binding Protein 1, MESH: Cells, Cultured, MESH: Cell Nucleus, MESH: Enzyme Activation, MESH : Cell Membrane, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Insulin, digestive system, 03 medical and health sciences, MESH: Sterol Regulatory Element Binding Protein 1, MESH : Sterol Regulatory Element Binding Protein 1, MESH: Blotting, Northern, Protein Precursors, Molecular Biology, Dose-Response Relationship, Drug, Glucokinase, Endoplasmic reticulum, Cell Membrane, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Sterol regulatory element-binding protein, Enzyme Activation, MESH: Phosphatidylinositol 3-Kinases, CCAAT-Enhancer-Binding Proteins, MESH: Female, Transcription Factors, MESH: Signal Transduction, Time Factors, MESH : Transcription Factors, MESH : Insulin, [ SDV.AEN ] Life Sciences [q-bio]/Food and Nutrition, MESH : Dose-Response Relationship, Drug, Endoplasmic Reticulum, MESH: Dose-Response Relationship, Drug, Phosphatidylinositol 3-Kinases, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], polycyclic compounds, MESH : Female, Cycloheximide, MESH: Cycloheximide, MESH : Protein Precursors, MESH : Endoplasmic Reticulum, MESH : Rats, [ SDV.MHEP.PHY ] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], MESH : Blotting, Northern, DNA-Binding Proteins, Fatty acid synthase, MESH : CCAAT-Enhancer-Binding Proteins, MESH: Enzyme Inhibitors, Female, lipids (amino acids, peptides, and proteins), MESH : Time Factors, Research Article, Signal Transduction, MESH: Rats, Immunoblotting, Biology, [ SDV.BC.IC ] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], [ SDV.BBM.MN ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN], MESH: Endoplasmic Reticulum, MESH : Cycloheximide, MESH : Cells, Cultured, medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, MESH : Protein Synthesis Inhibitors, Transcription factor, 030304 developmental biology, MESH : Signal Transduction, Cell Nucleus, MESH : Enzyme Inhibitors, MESH: Protein Synthesis Inhibitors, MESH: Transcription, Genetic, MESH: Time Factors, [ SDV.BC.BC ] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], MESH : Transcription, Genetic, MESH : Phosphatidylinositol 3-Kinases, Cell Biology, Blotting, Northern, Rats, biology.protein, Hepatocytes, MESH : Animals, MESH : Enzyme Activation, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, MESH: DNA-Binding Proteins, MESH: Cell Membrane
Abstract

International audience; The transcription factor sterol regulatory-element-binding protein-1c (SREBP-1c) plays a major role in the effect of insulin on the transcription of hepatic genes such as glucokinase and fatty acid synthase. We show here in cultured rat hepatocytes that insulin, through activation of the phosphatidylinositol 3-kinase pathway increases the abundance of the precursor form of SREBP-1c in endoplasmic reticulum. This precursor form is then rapidly cleaved, possibly irrespective of the continuous presence of insulin, leading to an increased content of the nuclear mature form of SREBP-1c. Nevertheless, the increased amount of the mature form of SREBP-1c in the nucleus is not a prerequisite for the rapid effect of insulin on the transcription of genes such as glucokinase, suggesting that additional actions of the hormone are involved, such as the activation of the nuclear form of SREBP-1c or of an unidentified SREBP-1c partner.

Published
2000

19. Increased cyclin D1 in vulnerable neurons in the hippocampus after ischaemia and epilepsy: a modulator of in vivo programmed cell death?

Author

Timsit, Serge, Rivera, Santiago, Ouaghi, Philippe, Guischard, Frédérique, Tremblay, Evelyne, Ben-Ari, Yehezkel, Khrestchatisky, Michel, Epilepsie et Ischemie Cerebrale, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Tyzio, Roman

Subjects
Male, MESH: Cyclin D1, MESH : RNA, Messenger, Time Factors, MESH: Hippocampus, MESH: Cyclin D2, MESH: Cyclin D3, MESH: Neurons, Gene Expression, Apoptosis, MESH: Cell Cycle, MESH : Hippocampus, Hippocampus, MESH: Nerve Degeneration, Brain Ischemia, MESH: Prosencephalon, MESH: Amygdala, Excitatory Amino Acid Agonists, Cyclin D2, Cyclin D1, MESH: Animals, Cyclin D3, In Situ Hybridization, Neurons, MESH : Cell Nucleus, Kainic Acid, MESH : Rats, MESH : Epilepsy, Cell Cycle, MESH : In Situ Hybridization, MESH: Brain Ischemia, Amygdala, MESH : Cyclin D1, MESH : Cyclin D2, MESH : Cyclin D3, MESH : Nerve Degeneration, MESH: Epilepsy, MESH : Time Factors, MESH: Cell Nucleus, MESH: Gene Expression, MESH: Rats, MESH : Male, MESH : Cyclins, Neurotoxins, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Rats, Wistar, MESH : Neurotoxins, MESH : Neurons, Prosencephalon, MESH: In Situ Hybridization, Cyclins, MESH : Cell Cycle, Animals, RNA, Messenger, Rats, Wistar, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MESH : Prosencephalon, MESH: Neurotoxins, MESH: RNA, Messenger, MESH : Excitatory Amino Acid Agonists, Cell Nucleus, Epilepsy, MESH : Amygdala, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Apoptosis, MESH: Time Factors, MESH: Biological Markers, MESH: Rats, Wistar, MESH: Kainic Acid, MESH: Cyclins, MESH: Male, Rats, MESH : Gene Expression, MESH : Biological Markers, MESH : Kainic Acid, nervous system, MESH : Brain Ischemia, Nerve Degeneration, MESH : Animals, MESH: Excitatory Amino Acid Agonists, MESH : Apoptosis, Biomarkers
Abstract

International audience; Several observations suggest that delayed neuronal death in ischaemia, epilepsy and other brain disorders includes an apoptotic component, involving programmed cell death (PCD). PCD is hypothesized to result, in part, from aberrant control of the cell cycle. Because they are instrumental in mitosis, cyclins D are key markers to evaluate whether neurons indeed progress into the cell cycle in situations of pathology. Therefore, we investigated in rat brains, the expression of cyclins D in the delayed neuronal death that occurs following transient global ischaemia and kainate-induced seizures. Following a four-vessel occlusion insult, quantitative in situ hybridization revealed a highly significant and persistent 100% increase of cyclin D1 mRNA in the vulnerable pyramidal neurons of the CA1 hippocampal region. Ischaemia also induced a smaller and transient cyclin D1 mRNA increase in the resistant CA3 area and dentate gyrus. In contrast, the cyclin D2 and D3 mRNAs, expressed constitutively in the adult rat hippocampus, were not upregulated. Following kainate-induced seizures, cyclin D1 mRNA was induced in the vulnerable CA3 region, and to a lesser extent, in non-vulnerable regions. Cyclin D1 immunohistochemistry revealed increased protein levels in the cytoplasm and nucleus of neurons commited to die after ischaemia. Double labelling experiments indicate that cyclin D1 is also expressed in reactive astrocytes but not in microglial cells. Finally, we report that in neurons, cyclin D1 expression peaks before nuclear condensation and the appearance of DNA fragmentation. We propose that cyclin D1, when expressed at high levels in lesioned neurons, may act as a modulator of apoptosis.

Published
1999

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