Author: "François Delalande" - Searchworks@Jio Institute Digital Library Search Results

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1. Zika virus enhances monocyte adhesion and transmigration favoring viral dissemination to neural cells

Author: Nilda Vanesa Ayala-Nunez, Gautier Follain, François Delalande, Aurélie Hirschler, Emma Partiot, Gillian L. Hale, Brigid C. Bollweg, Judith Roels, Maxime Chazal, Florian Bakoa, Margot Carocci, Sandrine Bourdoulous, Orestis Faklaris, Sherif R. Zaki, Anita Eckly, Béatrice Uring-Lambert, Frédéric Doussau, Sarah Cianferani, Christine Carapito, Frank M. J. Jacobs, Nolwenn Jouvenet, Jacky G. Goetz, and Raphael Gaudin
Subjects: Science
Abstract: Zika virus (ZIKV) can infect the central nervous system, but it is not clear how it reaches the brain. Here, Ayala-Nunez et al. show in ex vivo and in vivo models that ZIKV can hitch a ride in monocytes in a Trojan Horse manner to cross the endothelium and disseminate the virus.
Published: 2019
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2. Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes

Author: Shima Ghoroghi, Benjamin Mary, Annabel Larnicol, Nandini Asokan, Annick Klein, Naël Osmani, Ignacio Busnelli, François Delalande, Nicodème Paul, Sébastien Halary, Frédéric Gros, Laetitia Fouillen, Anne-Marie Haeberle, Cathy Royer, Coralie Spiegelhalter, Gwennan André-Grégoire, Vincent Mittelheisser, Alexandre Detappe, Kendelle Murphy, Paul Timpson, Raphaël Carapito, Marcel Blot-Chabaud, Julie Gavard, Christine Carapito, Nicolas Vitale, Olivier Lefebvre, Jacky G Goetz, and Vincent Hyenne
Subjects: exosome, Ral GTPase, pre-metastatic niche, Medicine, Science, Biology (General), QH301-705.5
Abstract: Cancer extracellular vesicles (EVs) shuttle at distance and fertilize pre-metastatic niches facilitating subsequent seeding by tumor cells. However, the link between EV secretion mechanisms and their capacity to form pre-metastatic niches remains obscure. Using mouse models, we show that GTPases of the Ral family control, through the phospholipase D1, multi-vesicular bodies homeostasis and tune the biogenesis and secretion of pro-metastatic EVs. Importantly, EVs from RalA or RalB depleted cells have limited organotropic capacities in vivoand are less efficient in promoting metastasis. RalA and RalB reduce the EV levels of the adhesion molecule MCAM/CD146, which favors EV-mediated metastasis by allowing EVs targeting to the lungs. Finally, RalA, RalB, and MCAM/CD146, are factors of poor prognosis in breast cancer patients. Altogether, our study identifies RalGTPases as central molecules linking the mechanisms of EVs secretion and cargo loading to their capacity to disseminate and induce pre-metastatic niches in a CD146-dependent manner.
Published: 2021
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3. Corrigendum: Morphine Binds Creatine Kinase B and Inhibits Its Activity

Author: Ivan Weinsanto, Jinane Mouheiche, Alexis Laux-Biehlmann, François Delalande, Arnaud Marquette, Virginie Chavant, Florian Gabel, Sarah Cianferani, Alexandre Charlet, Marie-Odile Parat, and Yannick Goumon
Subjects: morphine, complex, ligand-binding protein, creatine kinase, high affinity, Neurosciences. Biological psychiatry. Neuropsychiatry, RC321-571
Published: 2019
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4. Morphine Binds Creatine Kinase B and Inhibits Its Activity

Author: Ivan Weinsanto, Jinane Mouheiche, Alexis Laux-Biehlmann, François Delalande, Arnaud Marquette, Virginie Chavant, Florian Gabel, Sarah Cianferani, Alexandre Charlet, Marie-Odile Parat, and Yannick Goumon
Subjects: morphine, complex, ligand-binding protein, creatine kinase, high affinity, Neurosciences. Biological psychiatry. Neuropsychiatry, RC321-571
Abstract: Morphine is an analgesic alkaloid used to relieve severe pain, and irreversible binding of morphine to specific unknown proteins has been previously observed. In the brain, changes in the expression of energy metabolism enzymes contribute to behavioral abnormalities during chronic morphine treatment. Creatine kinase B (CK-B) is a key enzyme involved in brain energy metabolism. CK-B also corresponds to the imidazoline-binding protein I2 which binds dopamine (a precursor of morphine biosynthesis) irreversibly. Using biochemical approaches, we show that recombinant mouse CK-B possesses a μM affinity for morphine and binds to morphine in vitro. The complex formed by CK-B and morphine is resistant to detergents, reducing agents, heat treatment and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). CK-B-derived peptides CK-B1–75 and CK-B184–258 were identified as two specific morphine binding-peptides. In vitro, morphine (1–100 μM) significantly reduces recombinant CK-B enzymatic activity. Accordingly, in vivo morphine administration (7.5 mg/kg, i.p.) to mice significantly decreased brain extract CK-B activity compared to saline-treated animals. Together, these results show that morphine strongly binds CK-B and inhibits its activity in vitro and in vivo.
Published: 2018
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5. Pedagogia da criação musical hoje: partir da infância, passar pela adolescência e ir além

Author: François Delalande (Tradução de Tamya Moreira)
Subjects: composição (música), composição musical por computador, música e crianças, música na educação, Music and books on Music
Abstract: A criação musical, que há setenta anos estava restrita a músicos profissionais, é hoje acessível a amadores e crianças. Esta prática pode ser sua primeira experiência musical e a base de um processo educacional. O estudo de estratégias composicionais mostra que no centro do processo criativo encontra-se uma “ideia musical”. O que é uma “ideia musical”? Trata-se de uma singularidade sonora que atrai a atenção do músico e o incita a estendê- -la. Para estender sua descoberta, ele a repete com variações. A ideia musical – que há um século era geralmente um tema ou um motivo – é, nos dias de hoje, geralmente uma sonoridade particular. No entanto, repetir e variar um som distinto é típico do comportamento de exploração sonora da primeira infância, algo conhecido como “reação circular”. Assim, desde a infância, é possível encorajar um comportamento de exploração que se torna invenção, uma vez que seja intencional. Tal comportamento pode ser enriquecido pela dimensão simbólica e, posteriormente, pelo gosto pela construção. A criança percorre diferentes formas de jogo, como analisado por Piaget: o jogo sensório-motor, o jogo simbólico e o jogo baseado em regras. O papel do educador é, então, estimular e guiar este desenvolvimento espontâneo, e alguns equipamentos podem ajuda-lo. Por volta de nove ou dez anos de idade, o computador é uma ferramenta conveniente para a composição.
Published: 2017
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6. Na ausência da partitura: o caso singular da análise de música eletroacústica

Author: François Delalande
Subjects: Philosophy (General), B1-5802, Arts in general, NX1-820
Published: 2017

7. Cateslytin, a chromogranin A derived peptide is active against Staphylococcus aureus and resistant to degradation by its proteases.

Author: Rizwan Aslam, Céline Marban, Christian Corazzol, François Jehl, François Delalande, Alain Van Dorsselaer, Gilles Prévost, Youssef Haïkel, Corinne Taddei, Francis Schneider, and Marie-Hélène Metz-Boutigue
Subjects: Medicine, Science
Abstract: Innate immunity involving antimicrobial peptides represents an integrated and highly effective system of molecular and cellular mechanisms that protects host against infections. One of the most frequent hospital-acquired pathogens, Staphylococcus aureus, capable of producing proteolytic enzymes, which can degrade the host defence agents and tissue components. Numerous antimicrobial peptides derived from chromogranins, are secreted by nervous, endocrine and immune cells during stress conditions. These kill microorganisms by their lytic effect at micromolar range, using a pore-forming mechanism against Gram-positive bacteria, filamentous fungi and yeasts. In this study, we tested antimicrobial activity of chromogranin A-derived peptides (catestatin and cateslytin) against S. aureus and analysed S. aureus-mediated proteolysis of these peptides using HPLC, sequencing and MALDI-TOF mass spectrometry. Interestingly, this study is the first to demonstrate that cateslytin, the active domain of catestatin, is active against S. aureus and is interestingly resistant to degradation by S. aureus proteases.
Published: 2013
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8. The Pollen Coat Proteome: At the Cutting Edge of Plant Reproduction

Author: Juan David Rejón, François Delalande, Christine Schaeffer-Reiss, Juan de Dios Alché, María Isabel Rodríguez-García, Alain Van Dorsselaer, and Antonio Jesús Castro
Subjects: olive, pollen coat, proteomics, self-incompatibility, tapetum, Microbiology, QR1-502
Abstract: The tapetum is a single layer of secretory cells which encloses the anther locule and sustains pollen development and maturation. Upon apoptosis, the remnants of the tapetal cells, consisting mostly of lipids and proteins, fill the pits of the sculpted exine to form the bulk of the pollen coat. This extracellular matrix forms an impermeable barrier that protects the male gametophyte from water loss and UV light. It also aids pollen adhesion and hydration and retains small signaling compounds involved in pollen–stigma communication. In this study, we have updated the list of the pollen coat’s protein components and also discussed their functions in the context of sexual reproduction
Published: 2016
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9. Detection of prion protein in urine-derived injectable fertility products by a targeted proteomic approach.

Author: Alain Van Dorsselaer, Christine Carapito, François Delalande, Christine Schaeffer-Reiss, Daniele Thierse, Hélène Diemer, Douglas S McNair, Daniel Krewski, and Neil R Cashman
Subjects: Medicine, Science
Abstract: BackgroundIatrogenic transmission of human prion disease can occur through medical or surgical procedures, including injection of hormones such as gonadotropins extracted from cadaver pituitaries. Annually, more than 300,000 women in the United States and Canada are prescribed urine-derived gonadotropins for infertility. Although menopausal urine donors are screened for symptomatic neurological disease, incubation of Creutzfeldt-Jakob disease (CJD) is impossible to exclude by non-invasive testing. Risk of carrier status of variant CJD (vCJD), a disease associated with decades-long peripheral incubation, is estimated to be on the order of 100 per million population in the United Kingdom. Studies showing infectious prions in the urine of experimental animals with and without renal disease suggest that prions could be present in asymptomatic urine donors. Several human fertility products are derived from donated urine; recently prion protein has been detected in preparations of human menopausal gonadotropin (hMG).Methodology/principal findingsUsing a classical proteomic approach, 33 and 34 non-gonadotropin proteins were identified in urinary human chorionic gonadotropin (u-hCG) and highly-purified urinary human menopausal gonadotropin (hMG-HP) products, respectively. Prion protein was identified as a major contaminant in u-hCG preparations for the first time. An advanced prion protein targeted proteomic approach was subsequently used to conduct a survey of gonadotropin products; this approach detected human prion protein peptides in urine-derived injectable fertility products containing hCG, hMG and hMG-HP, but not in recombinant products.Conclusions/significanceThe presence of protease-sensitive prion protein in urinary-derived injectable fertility products containing hCG, hMG, and hMG-HP suggests that prions may co-purify in these products. Intramuscular injection is a relatively efficient route of transmission of human prion disease, and young women exposed to prions can be expected to survive an incubation period associated with a minimal inoculum. The risks of urine-derived fertility products could now outweigh their benefits, particularly considering the availability of recombinant products.
Published: 2011
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10. Endogenous morphine levels are increased in sepsis: a partial implication of neutrophils.

Author: Elise Glattard, Ingeborg D Welters, Thomas Lavaux, Arnaud H Muller, Alexis Laux, Dan Zhang, Alexander R Schmidt, François Delalande, Benoît-Joseph Laventie, Sylvie Dirrig-Grosch, Didier A Colin, Alain Van Dorsselaer, Dominique Aunis, Marie-Hélène Metz-Boutigue, Francis Schneider, and Yannick Goumon
Subjects: Medicine, Science
Abstract: BACKGROUND: Mammalian cells synthesize morphine and the respective biosynthetic pathway has been elucidated. Human neutrophils release this alkaloid into the media after exposure to morphine precursors. However, the exact role of endogenous morphine in inflammatory processes remains unclear. We postulate that morphine is released during infection and can be determined in the serum of patients with severe infection such as sepsis. METHODOLOGY: The presence and subcellular immunolocalization of endogenous morphine was investigated by ELISA, mass spectrometry analysis and laser confocal microscopy. Neutrophils were activated with Interleukin-8 (IL-8) or lipopolysaccharide (LPS). Morphine secretion was determined by a morphine-specific ELISA. mu opioid receptor expression was assessed with flow cytometry. Serum morphine concentrations of septic patients were determined with a morphine-specific ELISA and morphine identity was confirmed in human neutrophils and serum of septic patients by mass spectrometry analysis. The effects of the concentration of morphine found in serum of septic patients on LPS-induced release of IL-8 by human neutrophils were tested. PRINCIPAL FINDINGS: We confirmed the presence of morphine in human neutrophil extracts and showed its colocalisation with lactoferrin within the secondary granules of neutrophils. Morphine secretion was quantified in the supernatant of activated human polymorphonuclear neutrophils in the presence and absence of Ca(2+). LPS and IL-8 were able to induce a significant release of morphine only in presence of Ca(2+). LPS treatment increased mu opioid receptor expression on neutrophils. Low concentration of morphine (8 nM) significantly inhibited the release of IL-8 from neutrophils when coincubated with LPS. This effect was reversed by naloxone. Patients with sepsis, severe sepsis and septic shock had significant higher circulating morphine levels compared to patients with systemic inflammatory response syndrome and healthy controls. Mass spectrometry analysis showed that endogenous morphine from serum of patient with sepsis was identical to poppy-derived morphine. CONCLUSIONS: Our results indicate that morphine concentrations are increased significantly in the serum of patients with systemic infection and that morphine is, at least in part, secreted from neutrophils during sepsis. Morphine concentrations equivalent to those found in the serum of septic patients significantly inhibited LPS-induced IL-8 secretion in neutrophils.
Published: 2010
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11. Two chromogranin a-derived peptides induce calcium entry in human neutrophils by calmodulin-regulated calcium independent phospholipase A2.

Author: Dan Zhang, Peiman Shooshtarizadeh, Benoît-Joseph Laventie, Didier André Colin, Jean-François Chich, Jasmina Vidic, Jean de Barry, Sylvette Chasserot-Golaz, François Delalande, Alain Van Dorsselaer, Francis Schneider, Karen Helle, Dominique Aunis, Gilles Prévost, and Marie-Hélène Metz-Boutigue
Subjects: Medicine, Science
Abstract: BACKGROUND: Antimicrobial peptides derived from the natural processing of chromogranin A (CgA) are co-secreted with catecholamines upon stimulation of chromaffin cells. Since PMNs play a central role in innate immunity, we examine responses by PMNs following stimulation by two antimicrobial CgA-derived peptides. METHODOLOGY/PRINCIPAL FINDINGS: PMNs were treated with different concentrations of CgA-derived peptides in presence of several drugs. Calcium mobilization was observed by using flow cytometry and calcium imaging experiments. Immunocytochemistry and confocal microscopy have shown the intracellular localization of the peptides. The calmodulin-binding and iPLA2 activating properties of the peptides were shown by Surface Plasmon Resonance and iPLA2 activity assays. Finally, a proteomic analysis of the material released after PMNs treatment with CgA-derived peptides was performed by using HPLC and Nano-LC MS-MS. By using flow cytometry we first observed that after 15 s, in presence of extracellular calcium, Chromofungin (CHR) or Catestatin (CAT) induce a concentration-dependent transient increase of intracellular calcium. In contrast, in absence of extra cellular calcium the peptides are unable to induce calcium depletion from the stores after 10 minutes exposure. Treatment with 2-APB (2-aminoethoxydiphenyl borate), a store operated channels (SOCs) blocker, inhibits completely the calcium entry, as shown by calcium imaging. We also showed that they activate iPLA2 as the two CaM-binding factors (W7 and CMZ) and that the two sequences can be aligned with the two CaM-binding domains reported for iPLA2. We finally analyzed by HPLC and Nano-LC MS-MS the material released by PMNs following stimulation by CHR and CAT. We characterized several factors important for inflammation and innate immunity. CONCLUSIONS/SIGNIFICANCE: For the first time, we demonstrate that CHR and CAT, penetrate into PMNs, inducing extracellular calcium entry by a CaM-regulated iPLA2 pathway. Our study highlights the role of two CgA-derived peptides in the active communication between neuroendocrine and immune systems.
Published: 2009
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12. Endogenous morphine in SH-SY5Y cells and the mouse cerebellum.

Author: Arnaud Muller, Elise Glattard, Omar Taleb, Véronique Kemmel, Alexis Laux, Monique Miehe, François Delalande, Guy Roussel, Alain Van Dorsselaer, Marie-Hélène Metz-Boutigue, Dominique Aunis, and Yannick Goumon
Subjects: Medicine, Science
Abstract: Morphine, the principal active agent in opium, is not restricted to plants, but is also present in different animal tissues and cell types, including the mammalian brain. In fact, its biosynthetic pathway has been elucidated in a human neural cell line. These data suggest a role for morphine in brain physiology (e.g., neurotransmission), but this hypothesis remains a matter of debate. Recently, using the adrenal neuroendocrine chromaffin cell model, we have shown the presence of morphine-6-glucuronide (M6G) in secretory granules and their secretion products, leading us to propose that these endogenous alkaloids might represent new neuroendocrine factors. Here, we investigate the potential function of endogenous alkaloids in the central nervous system.Microscopy, molecular biology, electrophysiology, and proteomic tools were applied to human neuroblastoma SH-SY5Y cells (i) to characterize morphine and M6G, and (ii) to demonstrate the presence of the UDP-glucuronyltransferase 2B7 enzyme, which is responsible for the formation of M6G from morphine. We show that morphine is secreted in response to nicotine stimulation via a Ca(2+)-dependent mechanism involving specific storage and release mechanisms. We also show that morphine and M6G at concentrations as low as 10(-10) M are able to evoke specific naloxone-reversible membrane currents, indicating possible autocrine/paracrine regulation in SH-SY5Y cells. Microscopy and proteomic approaches were employed to detect and quantify endogenous morphine in the mouse brain. Morphine is present in the hippocampus, cortex, olfactory bulb, and cerebellum at concentration ranging from 1.45 to 7.5 pmol/g. In the cerebellum, morphine immunoreactivity is localized to GABA basket cells and their termini, which form close contacts on Purkinje cell bodies.The presence of morphine in the brain and its localization in particular areas lead us to conclude that it has a specific function in neuromodulation and/or neurotransmission. Furthermore, its presence in cerebellar basket cell termini suggests that morphine has signaling functions in Purkinje cells that remain to be discovered.
Published: 2008
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13. The Holdup Multiplex, an assay for high-throughput measurement of protein-ligand affinity constants using a mass-spectrometry readout

Author: François Delalande, Gergo Gogl, Aurélien Rohrbacher, Camille Kostmann, Pascal Eberling, Christine Carapito, Gilles Travé, and Elodie Monsellier
Abstract: The accurate description and subsequent modeling of protein interactomes requires quantification of their affinities at proteome-wide scale. Here we develop and validate the Holdup Multiplex, a versatile assay for high-throughput measurement of protein-ligand affinity constants that uses mass-spectrometry as readout. The method can quantify thousands of affinities in one single run, with high precision and over several orders of magnitude. We applied this strategy to the seven human 14-3-3 isoforms, quantifying in a few sample-runs their interaction with 1,000 different phosphopeptides. We were able to identify hundreds of new 14-3-3 binding sites. We showed that the seven human 14-3-3 display similar specificities but staggered affinities, 14-3-3g being always the best binder and 14-3-3ε and σ, the weakest. Finally, we identified dozens of 14-3-3 bindings sites, some intervening in key signaling pathways, that were either stabilized or destabilized by the phytotoxin Fusicoccin-A. Our approach, which throughput can be pushed up to the sensitivity limit of the mass-spectrometry setup, is applicable to any category of protein-ligand interactions and thus bears a wide potential both for high-throughput interactomics and chemoproteomics.
Published: 2022

14. Surface charge influences protein corona, cell uptake and biological effects of carbon dots

Author: Yasmin Arezki, François Delalande, Christine Schaeffer-Reiss, Sarah Cianférani, Mickaël Rapp, Luc Lebeau, Françoise Pons, and Carole Ronzani
Subjects: Proteomics, Surface Properties, Aucun, Carbon, Citric Acid, Albumins, Nanoparticles, Protein Corona, General Materials Science, Adiponectin, Vitronectin, Amines, Apolipoproteins C, Fetuins, Apolipoproteins B
Abstract: Carbon dots are emerging nanoparticles (NPs) with tremendous applications, especially in the biomedical field. Herein is reported the first quantitative proteomic analysis of the protein corona formed on CDs with different surface charge properties. Four CDs were synthesized from citric acid and various amine group-containing passivation reagents, resulting in cationic NPs with increasing zeta (ζ)-potential and density of positive charges. After CD contact with serum, we show that protein corona identity is influenced by CD surface charge properties, which in turn impacts CD uptake and viability loss in macrophages. In particular, CDs with high ζ-potential (+30 mV) and charge density (2 μmol mg
Published: 2022

15. Trans-synaptic dwelling of SARS-CoV-2 particles perturbs neural synapse organization and function

Author: Emma Partiot, Aurélie Hirschler, Sophie Colomb, Willy Lutz, Tine Claeys, François Delalande, Maika S. Deffieu, Judith R.E. Roels, Joanna Bons, Domitille Callon, Laurent Andreoletti, Marc Labrousse, Frank M.J. Jacobs, Valérie Rigau, Benoit Charlot, Lennart Martens, Christine Carapito, Gowrishankar Ganesh, and Raphael Gaudin
Abstract: SARS-CoV-2 infection is associated with short- and long-term neurological and psychiatric complications, referred to as neuroCOVID. These symptoms are relatively heterogenous and fluctuating, hampering the discovery of molecular mechanisms underlying viro-induced brain perturbations. Here, we show that the human cerebral cortex poorly supports SARS-CoV-2 dissemination using post-mortem COVID-19 patient samples, ex vivo organotypic cultures of human brain explants and stem cell-derived cortical organoids. Despite restricted infection, the sole exposure of neural cells to SARS-CoV-2 particles is sufficient to induce significant perturbations on neural synapse organization associated to electrical activity dysfunction. Single-organoid proteomics revealed that exposure to SARS-CoV-2 is associated to trans-synaptic proteins upregulation and unveiled that incoming virions dwell at LPHN3/FLRT3-containing synapses. Our study provides new mechanistic insights on the origin of SARS-CoV-2-induced neurological disorders.One-Sentence SummarySARS-CoV-2 modulates neural plasticity and electrical activity as viral particles lodge at the trans-synaptic interface.
Published: 2022

16. The Emergence of the Metabolic Signaling of the Nucleoredoxin-like Genes during Evolution

Author: Najate Aït-Ali, Frédéric Blond, Emmanuelle Clérin, Ala Morshedian, Quénol Cesar, François Delalande, Mitsumasa Koyanagi, Catherine Birck, John Han, Xiaoyuan Ren, Alain van Dorsselaer, Akihisa Terakita, Gordon L. Fain, and Thierry Léveillard
Subjects: genetic structures, sense organs
Abstract: SUMMARYThe nucleoredoxin-like genes NXNL1 and NXNL2 were identified through the biological activity of rod-derived cone viability factors (RdCVF and RdCVF2), the alternatively spliced variants produced by intron retention, that mediate signaling between rod and cone photoreceptors by stimulating glucose uptake. These therapeutic genes for inherited retinal degenerations also produce by splicing thioredoxin-like proteins that reduce oxidized cysteines in photoreceptor proteins. The first NXNL genes date from the first animal phyla. Intron retention produces an active RdCVF protein in the tentacles of Hydra vulgaris, a species without eyes. A Scallop RdCVF protein is produced by ciliated photoreceptors of the retina and binds its receptor, BSG1. In the lamprey, a descendent of early vertebrates, RdCVF metabolic signaling between rod and cones is fully established. In the mouse, the production of BSG1 by photoreceptors is regulated by cell-specific splicing inhibition. RdCVF signaling predates photoreceptors and evolved through two alternative splicing events.
Published: 2022

17. The invention of sound

Author: François Delalande
Subjects: geography, geography.geographical_feature_category, Computer science, Acoustics, General Medicine, Sound (geography)
Published: 2020

18. Combining label-free and label-based accurate quantifications with SWATH-MS: Comparison with SRM and PRM for the evaluation of bovine muscle type effects

Author: Brigitte Picard, Frédéric Bertrand, Joanna Bons, Muriel Bonnet, Christine Carapito, Sarah Cianférani, Marie Chion, François Delalande, Gauthier Husson, Myriam Maumy-Bertrand, Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche Mathématique Avancée (IRMA), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche sur les Herbivores - UMR 1213 (UMRH), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire Modélisation et Sûreté des Systèmes (LM2S), Laboratoire Informatique et Société Numérique (LIST3N), Université de Technologie de Troyes (UTT)-Université de Technologie de Troyes (UTT), French Ministry of Science INRAE-Phase Division (RefQuant project) French Proteomic Infrastructure (ProFI) ANR-10-INBS-08-03, Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Université Clermont Auvergne (UCA), Institut Charles Delaunay (ICD), Université de Technologie de Troyes (UTT)-Centre National de la Recherche Scientifique (CNRS)-Université de Technologie de Troyes (UTT)-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)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Technologie de Troyes (UTT), and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)
Subjects: Swath ms, Proteome, Computer science, [SDV]Life Sciences [q-bio], Marbled meat, Computational biology, Mass spectrometry, Biochemistry, Mass Spectrometry, 03 medical and health sciences, Meat tenderness, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.IDA]Life Sciences [q-bio]/Food engineering, Animals, Humans, Bovine muscle, Beef meat quality, Targeted proteomics, Molecular Biology, 030304 developmental biology, Label free, 0303 health sciences, Muscles, 030302 biochemistry & molecular biology, Selected reaction monitoring, Quantitative MS, SWATH-MS, Cattle, Data-Independent Acquisition (DIA), Biomarkers
Abstract: International audience; Mass spectrometry has proven to be a valuable tool for the accurate quantification of proteins. In this study, the performances of three targeted approaches, namely selected reaction monitoring (SRM), parallel reaction monitoring (PRM) and sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS), to accurately quantify ten potential biomarkers of beef meat tenderness or marbling in a cohort of 64 muscle samples were evaluated. So as to get the most benefit out of the complete MS2 maps that are acquired in SWATH-MS, an original label-free quantification method to estimate protein amounts using an I-spline regression model was developed. Overall, SWATH-MS outperformed SRM in terms of sensitivity and dynamic range, while PRM still performed the best, and all three strategies showed similar quantification accuracies and precisions for the absolute quantification of targets of interest. This targeted picture was extended by 585 additional proteins for which amounts were estimated using the label-free approach on SWATH-MS; thus, offering a more global profiling of muscle proteomes and further insights into muscle type effect on candidate biomarkers of beef meat qualities as well as muscle metabolism.
Published: 2021

19. Rab7-harboring vesicles are carriers of the transferrin receptor through the biosynthetic secretory pathway

Author: Cristina M. Dorobantu, Vincent Lucansky, Franck Perez, Christine Carapito, Gaelle Boncompain, François Delalande, Maïka Deffieu, Raphael Gaudin, Ieva Cesonyte, Sarah Cianférani, Eli Song, Aurélie Hirschler, Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg Inserm Faculté de médecine, Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut Curie [Paris], Biologie Cellulaire et Cancer, and Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0303 health sciences, Multidisciplinary, Chemistry, Vesicle, [SDV]Life Sciences [q-bio], SciAdv r-articles, Life Sciences, Transferrin receptor, Compartment (chemistry), Cell Biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Membrane transport, Secretory Vesicle, 3. Good health, Cell biology, 03 medical and health sciences, 0302 clinical medicine, RAB7A, 030217 neurology & neurosurgery, Intracellular, Secretory pathway, Research Articles, 030304 developmental biology, Research Article
Abstract: CRISPR-based bioengineering of the transferrin receptor reveals a new role for Rab7 in the biosynthetic secretory pathway., The biosynthetic secretory pathway is particularly challenging to investigate as it is underrepresented compared to the abundance of the other intracellular trafficking routes. Here, we combined the retention using selective hook (RUSH) to a CRISPR-Cas9 gene editing approach (eRUSH) and identified Rab7-harboring vesicles as an important intermediate compartment of the Golgi–to–plasma membrane transport of neosynthesized transferrin receptor (TfR). These vesicles did not exhibit degradative properties and were not associated to Rab6A-harboring vesicles. Rab7A was transiently associated to neosynthetic TfR-containing post–Golgi vesicles but dissociated before fusion with the plasma membrane. Together, our study reveals a role for Rab7 in the biosynthetic secretory pathway of the TfR, highlighting the diversity of the secretory vesicles’ nature.
Published: 2021

20. Author response: Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes

Author: Coralie Spiegelhalter, Frédéric Gros, Anne-Marie Haeberlé, Ignacio Busnelli, Christine Carapito, Vincent Mittelheisser, Nicolas Vitale, Jacky G. Goetz, Marcel Blot-Chabaud, Julie Gavard, Laetitia Fouillen, Paul Timpson, Nandini Asokan, Gwennan André-Grégoire, Alexandre Detappe, Raphael Carapito, Benjamin Mary, Olivier Lefebvre, Naël Osmani, Shima Ghoroghi, Kendelle J. Murphy, Nicodème Paul, Annabel Larnicol, Vincent Hyenne, Annick Klein, Catherine A. Royer, Sébastien Halary, and François Delalande
Subjects: business.industry, Cancer research, Medicine, Breast cancer metastasis, GTPase, business, Microvesicles, Biogenesis
Published: 2020

21. Ral GTPases promote metastasis by controlling biogenesis and organ colonization of exosomes

Author: Marcel Blot-Chabaud, François Delalande, Shima Ghoroghi, Nicolas Vitale, Annick Klein, Laetitia Fouillen, Benjamin Mary, Coralie Spiegelhalter, Sébastien Halary, Nicodème Paul, Naël Osmani, Annabel Larnicol, Ignacio Busnelli, Jacky G. Goetz, Frédéric Gros, Julie Gavard, Anne-Marie Haeberlé, Raphael Carapito, Catherine A. Royer, Paul Timpson, Olivier Lefebvre, Gwennan André-Grégoire, Kendelle J. Murphy, Vincent Hyenne, Christine Carapito, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg (UNISTRA), Immuno-Rhumatologie Moléculaire, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Molécules de Communication et Adaptation des Micro-organismes (MCAM), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de biogenèse membranaire (LBM), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS), Institut des Neurosciences Cellulaires et Intégratives (INCI), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre de Neurochimie, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre recherche en CardioVasculaire et Nutrition = Center for CardioVascular and Nutrition research (C2VN), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Gavard, Julie, and Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0303 health sciences, RALB, Stromal cell, [SDV]Life Sciences [q-bio], GTPase, Biology, medicine.disease, Microvesicles, RALA, Metastasis, Cell biology, [SDV] Life Sciences [q-bio], 03 medical and health sciences, 0302 clinical medicine, 030220 oncology & carcinogenesis, medicine, CD146, Secretion, 030304 developmental biology
Abstract: International audience; Cancer extracellular vesicles (EVs) mainly exert pro-tumoral functions by changing the phenotypes of stromal cells to the benefit of tumor growth and metastasis. They shuttle to distant organs and fertilize pre-metastatic niches facilitating subsequent seeding by circulating tumor cells. The levels of tumor secreted EVs correlate with tumor aggressiveness, however, the link between EV secretion mechanisms and their capacity to form pre-metastatic niches remains obscure. Here, we show that GTPases of the Ral family control, through the phospholipase D1, multi-vesicular bodies homeostasis and thereby tune the biogenesis and secretion of pro-metastatic EVs. RalA and RalB promote lung metastasis in a syngeneic mouse model. Importantly, EVs from RalA or RalB depleted cells have limited organotropic capacities in vivo and, as a consequence, are less efficient in promoting lung metastasis. RalA or RalB modulate the EV levels of the adhesion molecule MCAM/CD146, which mediates lung col-onization. Finally, RalA and RalB, but also MCAM/CD146, are factors of poor prognosis in human breast cancer patients. Altogether , our study identifies Ral GTPases as central molecules linking the mechanisms of EVs secretion, cargo loading to their capacity to disseminate and induce pre-metastatic niches.
Published: 2020

22. CRISPR-based bioengineering of the Transferrin Receptor revealed a role for Rab7 in the biosynthetic secretory pathway

Author: Raphael Gaudin, Christine Carapito, Cristina M. Dorobantu, François Delalande, Eli Song, Ieva Cesonyte, Gaelle Boncompain, Maïka Deffieu, Tao Xu, Sarah Cianférani, Franck Perez, Vincent Lucansky, and Aurélie Hirschler
Subjects: Vesicle fusion, RAB7A, Chemistry, Vesicle, Quantitative proteomics, Transferrin receptor, Rab, Secretory Vesicle, Secretory pathway, Cell biology
Abstract: The regulated secretory trafficking of neosynthesized transmembrane receptors is particularly challenging to investigate as it is under-represented at steady state compared to the abundance of the other trafficking routes. Here, we combined the retention using selective hook (RUSH) system to a CRISPR/Cas9 gene editing approach (eRUSH) to identify molecular players involved in the trafficking of neosynthesized Transferrin Receptor (TfR) en route to the plasma membrane (PM). TfR-eRUSH monoclonal cells expressing endogenous, ER-retainable and fluorescent TfR were engineered and characterized. Spatiotemporal quantitative proteomics of TfR-eRUSH cells allowed the identification of molecular partners associated with TfR-containing membranes and provided a comprehensive list of potential regulators, co-trafficking cargos, and enriched pathways. Furthermore, we chose to focus our attention on the Rab GTPase family members for their function as vesicle trafficking regulators and performed a Rab-targeted siRNA screen that we correlated to our proteomics data. Unexpectedly, we identified Rab7-harboring vesicles as an intermediate compartment of the Golgi-to-PM transport of the neosynthetic TfR. These vesicles did not exhibit degradative properties and were not associated to Rab6A-harboring vesicles, also involved in Golgi-to-PM transport. However, Rab6A-TfR vesicles delivered TfR directly to the PM, while in contrast, Rab7A was transiently associated to neosynthetic TfR-containing post-Golgi vesicles but dissociated before PM vesicle fusion. Together, our study proposes the eRUSH as a powerful tool to further study the secretory pathway and reveals an unforeseen role for Rab7 in the neosynthetic transport of the TfR, highlighting the diversity of the secretory vesicles’ nature for a given cargo.
Published: 2020
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23. Stable isotope-labelled morphine to study in vivo central and peripheral morphine glucuronidation and brain transport in tolerant mice

Author: Pierrick Poisbeau, Marc Lamshöft, François Delalande, Jinane Mouheiche, Pascal Darbon, Marie-Odile Parat, Florian Gabel, Yannick Goumon, Virginie Chavant, Ivan Weinsanto, Sarah Cianférani, Alain Van Dorsselaer, Alexis Laux-Biehlmann, Tando Maduna, and Alexandre Charlet
Subjects: 0301 basic medicine, Pharmacology, Chemistry, Metabolite, Central nervous system, Analgesic, Glucuronidation, Metabolism, 3. Good health, 03 medical and health sciences, chemistry.chemical_compound, 030104 developmental biology, 0302 clinical medicine, medicine.anatomical_structure, Drug tolerance, In vivo, medicine, Morphine, 030217 neurology & neurosurgery, medicine.drug
Abstract: Chronic administration of medication can have an important impact on metabolic enzymes leading to physiological adaptations. Morphine metabolism in the liver has been extensively studied following acute morphine treatment but morphine metabolic processes in the central nervous system are poorly characterised. Long-term morphine treatment is limited by the development of tolerance, resulting in a decrease of its analgesic effect. Whether or not morphine analgesic tolerance affects in vivo brain morphine metabolism and blood-brain barrier (BBB) permeability remains a major pending question. Thus, our aim was to characterise the in vivo metabolism and BBB permeability of morphine after long-term treatment at both central and peripheral levels. Mice were injected with morphine or saline solution for 8 consecutive days in order to induce morphine analgesic tolerance. On the ninth day, both groups received a final injection of morphine (85%) and d3-morphine (morphine bearing three H; 15%, w/w). Mice were then euthanized and blood, urine, brain and liver samples were collected. LC-MS/MS was used to quantify morphine, its metabolite morphine-3-glucuronide (M3G) and their respective d3-labelled counterparts. We found no significant differences in morphine CNS uptake and metabolism between control and tolerant mice. This suggests that morphine analgesic tolerance is not linked to an increase of morphine glucuronidation into M3G or an alteration of the drug's global BBB permeability. Interestingly, d3-morphine metabolism was decreased compared to morphine without any interference with our study.
Published: 2018

24. Zika virus enhances monocyte adhesion and transmigration favoring viral dissemination to neural cells

Author: Emma Partiot, Nilda Vanesa Ayala-Nunez, Jacky G. Goetz, Gautier Follain, Judith R.E. Roels, Anita Eckly, Béatrice Uring-Lambert, Maxime Chazal, Raphael Gaudin, Gillian Hale, Frank M. J. Jacobs, Sandrine Bourdoulous, Orestis Faklaris, Sherif R. Zaki, Aurélie Hirschler, Nolwenn Jouvenet, Margot Carocci, Florian Bakoa, Sarah Cianférani, Christine Carapito, Frédéric Doussau, Brigid C. Bollweg, François Delalande, Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA), Institut National de la Santé et de la Recherche Médicale (INSERM), Immuno-Rhumatologie Moléculaire, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Strasbourg (UNISTRA), Fédération de Médecine Translationnelle de Strasbourg (FMTS), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centers for Disease Control and Prevention [Atlanta] (CDC), Centers for Disease Control and Prevention, Swammerdam Institute for Life Sciences, University of Amsterdam [Amsterdam] (UvA)-Center for Neurosciences, Génomique virale et vaccination, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université Paris Descartes - Paris 5 (UPD5), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), BioCampus Montpellier (BCM), Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Biologie et pharmacologie des plaquettes sanguines: hémostase, thrombose, transfusion, Université de Strasbourg (UNISTRA)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Les Hôpitaux Universitaires de Strasbourg (HUS), Institut des Neurosciences Cellulaires et Intégratives (INCI), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), This work has received funding from the French State via the French National Research Agency (ANR) as part of the program Investissements d’avenir (IdEx Université de Strasbourg) to RG. This work was supported by an ATIP-AVENIR starting grant to RG. The research leading to these results has received funding from the People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program (FP7/2007–2013) under REA grant agreement no. PCOFUND-GA-2013-609102, through the PRESTIGE program coordinated by Campus France, and from the French Agency for Research on AIDS and Viral Hepatitis (ANRS), both attributed to NVAN. Experiments conducted by G.F. and J.G.G. were funded by Plan Cancer (OptoMetaTrap), Ligue contre le Cancer, Idex Attractivités (University of Strasbourg), and by institutional funds from INSERM and University of Strasbourg. G.F. was supported by a doctoral fellowship from Ligue Contre le Cancer. Mass spectrometry experiments were supported by the French Proteomic Infrastructure (ProFI, ANR-10-INBS-08-03). F.B. is a recipient of a PhD grant provided by ANRT (Association Nationale de la Recherche et de la Technologie). N.J. is funded by Agence Nationale pour la Recherche (ANR-16-CE15-0025-01), Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur. CDC contributions to this paper written and edited by G.L.H. and B.C.B. in their private capacity. No official support or endorsement by the Centers for Disease Control and Prevention, Department of Health and Human Services is intended, nor should be inferred. All authors read, provided feedback, and approved the paper. The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 NL4-3 AD8 Infectious Molecular Clone (pNL(AD8)) from Dr. Eric O. Freed (cat# 11346)., ANR-16-CE15-0025,Viro-Storm,Mécanismes de production incontrôlée de cytokines au cours de l'infection virale(2016), European Project: 609102,EC:FP7:PEOPLE,FP7-PEOPLE-2013-COFUND,PRESTIGE(2014), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-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 Cité (UPCité), BioCampus (BCM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), EFS-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), jouvenet, nolwenn, Mécanismes de production incontrôlée de cytokines au cours de l'infection virale - - Viro-Storm2016 - ANR-16-CE15-0025 - AAPG2016 - VALID, PRES Towards International Gain of Excellence - PRESTIGE - - EC:FP7:PEOPLE2014-09-01 - 2019-08-31 - 609102 - VALID, Equipe Direction scientifique, Sciences et Technologies de la Musique et du Son (STMS), Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS), École pratique des hautes études (EPHE), Neuro-Immunologie Virale, Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Photonique Quantique et Moléculaire (LPQM), Centre National de la Recherche Scientifique (CNRS)-CentraleSupélec-École normale supérieure - Cachan (ENS Cachan), Biologie et pharmacologie des plaquettes sanguines: hémostase, thrombose, transfusion (http://www.u949.inserm.fr/), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS, Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse BioOrganique, Département des Sciences Analytiques, Institut Pluridisciplinaire Hubert Curien, Strasbourg, France (LSMBO-DSA-IPHC), Centre National de la Recherche Scientifique (CNRS), Immunorhumathologie moléculaire, Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), and Molecular Neuroscience (SILS, FNWI)
Subjects: Central Nervous System, 0301 basic medicine, MESH: Neurons, General Physics and Astronomy, MESH: Organoids, MESH: Monocytes, Monocytes, Zika virus, 0302 clinical medicine, MESH: Transendothelial and Transepithelial Migration, Cerebellum, MESH: Animals, lcsh:Science, MESH: Embryonic Stem Cells, ComputingMilieux_MISCELLANEOUS, Zebrafish, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Neurons, Multidisciplinary, Zika Virus Infection, Cell adhesion molecule, food and beverages, Sciences du Vivant [q-bio]/Microbiologie et Parasitologie, Phenotype, 3. Good health, Cell biology, Organoids, Mechanisms of disease, medicine.anatomical_structure, MESH: Cell Survival, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Female, Endothelium, Cell Survival, Science, MESH: Zika Virus, Biology, MESH: Endothelium, Article, General Biochemistry, Genetics and Molecular Biology, Virus, MESH: Cell Adhesion, 03 medical and health sciences, Virology, medicine, Animals, Humans, MESH: Central Nervous System, Cellular microbiology, Cell adhesion, MESH: Zebrafish, Embryonic Stem Cells, MESH: Humans, Monocyte, fungi, Transendothelial and Transepithelial Migration, Zika Virus, General Chemistry, biology.organism_classification, Embryonic stem cell, MESH: Cerebellum, Disease Models, Animal, 030104 developmental biology, lcsh:Q, MESH: Disease Models, Animal, Cell Adhesion Molecules, MESH: Female, 030217 neurology & neurosurgery
Abstract: Zika virus (ZIKV) invades and persists in the central nervous system (CNS), causing severe neurological diseases. However the virus journey, from the bloodstream to tissues through a mature endothelium, remains unclear. Here, we show that ZIKV-infected monocytes represent suitable carriers for viral dissemination to the CNS using human primary monocytes, cerebral organoids derived from embryonic stem cells, organotypic mouse cerebellar slices, a xenotypic human-zebrafish model, and human fetus brain samples. We find that ZIKV-exposed monocytes exhibit higher expression of adhesion molecules, and higher abilities to attach onto the vessel wall and transmigrate across endothelia. This phenotype is associated to enhanced monocyte-mediated ZIKV dissemination to neural cells. Together, our data show that ZIKV manipulates the monocyte adhesive properties and enhances monocyte transmigration and viral dissemination to neural cells. Monocyte transmigration may represent an important mechanism required for viral tissue invasion and persistence that could be specifically targeted for therapeutic intervention., Zika virus (ZIKV) can infect the central nervous system, but it is not clear how it reaches the brain. Here, Ayala-Nunez et al. show in ex vivo and in vivo models that ZIKV can hitch a ride in monocytes in a Trojan Horse manner to cross the endothelium and disseminate the virus.
Published: 2019

25. Corrigendum: Morphine Binds Creatine Kinase B and Inhibits Its Activity

Author: Arnaud Marquette, François Delalande, Sarah Cianférani, Florian Gabel, Alexis Laux-Biehlmann, Marie-Odile Parat, Yannick Goumon, Virginie Chavant, Alexandre Charlet, Ivan Weinsanto, and Jinane Mouheiche
Subjects: Research groups, biology, Drug discovery, business.industry, creatine kinase, morphine, high affinity, Ligand Binding Protein, Pharmacology, ligand-binding protein, lcsh:RC321-571, Cellular and Molecular Neuroscience, Creatine Kinase-B, biology.protein, Morphine, medicine, Creatine kinase, business, lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry, complex, medicine.drug
Abstract: In the published article, there was an error in affiliation "2." Instead of "Global Drug Discovery, Global Therapeutic Research Groups, Gynecological Therapies, Bayer Healthcare, Berlin, Germany", it should be "Laboratoire de Spectrometrie deMasse BioOrganique, IPHC-DSA, CNRS UMR7178 and Universite de Strasbourg, Strasbourg, France." Additionally, the current address of one author has been added as a footnote and is denoted using. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.
Published: 2019

26. Studying the Fate of Tumor Extracellular Vesicles at High Spatiotemporal Resolution Using the Zebrafish Embryo

Author: Jack Bauer, Benjamin Mary, Christine Carapito, Vincent Hyenne, Luc Mercier, Joanna Bons, François Delalande, Susana Garcia Silva, Frederik J. Verweij, Ana Amor López, Héctor Peinado, Jacky G. Goetz, Nina Fekonja, Mayeul Collot, Sébastien Harlepp, Farida Djouad, Andrey S. Klymchenko, Ignacio Busnelli, Olivier Lefebvre, Maria Jesus Garcia-Leon, Pedro Machado, Shima Ghoroghi, Guillaume van Niel, Gautier Follain, Immuno-Rhumatologie Moléculaire, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA)), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), European Molecular Biology Laboratory [Heidelberg] (EMBL), Compartimentation et dynamique cellulaires (CDC), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Instituto de Investigaciones Biomedicas, Universidad Nacional Autónoma de México (UNAM), Immunorhumathologie moléculaire, Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS), Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Institut Curie-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), and UMR1183, IRMB, Hôpital Saint-Eloi
Subjects: Stromal cell, [SDV]Life Sciences [q-bio], [SDV.CAN]Life Sciences [q-bio]/Cancer, Cell Communication, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Exosomes, Extracellular vesicles, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Extracellular Vesicles, 0302 clinical medicine, Animal model, Neoplasms, Tumor Microenvironment, Animals, Molecular Biology, Zebrafish, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Endothelial Cells, Cell Biology, biology.organism_classification, Microvesicles, Cell biology, Disease Models, Animal, Tumor progression, Zebrafish embryo, Disease Progression, Spatiotemporal resolution, Stromal Cells, 030217 neurology & neurosurgery, Developmental Biology
Abstract: International audience; Tumor extracellular vesicles (EVs) mediate the communication between tumor and stromal cells mostly to the benefit of tumor progression. Notably, tumor EVs travel in the bloodstream, reach distant organs, and locally modify the microenvironment. However, visualizing these events in vivo still faces major hurdles. Here, we describe an approach for tracking circulating tumor EVs in a living organism: we combine chemical and genetically encoded probes with the zebrafish embryo as an animal model. We provide a first description of tumor EVs’ hemodynamic behavior and document their intravascular arrest. We show that circulating tumor EVs are rapidly taken up by endothelial cells and blood patrolling macrophages and subsequently stored in degradative compartments. Finally, we demonstrate that tumor EVs activate macrophages and promote metastatic outgrowth. Overall, our study proves the usefulness and prospects of zebrafish embryo to track tumor EVs and dissect their role in metastatic niches formation in vivo.
Published: 2019

27. Stable isotope-labelled morphine to study in vivo central and peripheral morphine glucuronidation and brain transport in tolerant mice

Author: Ivan, Weinsanto, Alexis, Laux-Biehlmann, Jinane, Mouheiche, Tando, Maduna, François, Delalande, Virginie, Chavant, Florian, Gabel, Pascal, Darbon, Alexandre, Charlet, Pierrick, Poisbeau, Marc, Lamshöft, Alain, Van Dorsselaer, Sarah, Cianferani, Marie-Odile, Parat, Yannick, Goumon, Institut des Neurosciences Cellulaires et Intégratives (INCI), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), UNIVERSITY OF TECHNOLOGY DORTMUND DEU, Partenaires IRSTEA, Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), University of Queensland [Brisbane], Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Équipe 'Rythme, vie et mort de la rétine', Université de Strasbourg (UNISTRA)-Institut des Neurosciences Cellulaires et Intégratives (INCI)-Centre National de la Recherche Scientifique (CNRS), Département Neurotransmission et sécrétion neuroendocrine, Equipe Direction scientifique, Sciences et Technologies de la Musique et du Son (STMS), Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse BioOrganique, Département des Sciences Analytiques, Institut Pluridisciplinaire Hubert Curien (LSMBO-DSA-IPHC), Centre National de la Recherche Scientifique (CNRS), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Charlet, Alexandre, Plateau technique de Spectrométrie de Masse (SM-INCI / CNRS UPR3212), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Technische Universität Dortmund [Dortmund] (TU), and Outcomes Research Consortium [Cleveland, OH, USA] (ORC)
Subjects: Male, Sciences du Vivant [q-bio]/Neurosciences [q-bio.NC], Morphine, [SCCO.NEUR]Cognitive science/Neuroscience, [SCCO.NEUR] Cognitive science/Neuroscience, Molecular Conformation, Brain, Drug Tolerance, Research Papers, Mice, Inbred C57BL, Mice, Glucuronides, Isotope Labeling, Animals, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Cells, Cultured, ComputingMilieux_MISCELLANEOUS
Abstract: International audience; Background and purpose: Chronic administration of medication can significantly affect metabolic enzymes leading to physiological adaptations. Morphine metabolism in the liver has been extensively studied following acute morphine treatment, but such metabolic processes in the CNS are poorly characterized. Long-term morphine treatment is limited by the development of tolerance, resulting in a decrease of its analgesic effect. Whether or not morphine analgesic tolerance affects in vivo brain morphine metabolism and blood-brain barrier (BBB) permeability remains a major question. Here, we have attempted to characterize the in vivo metabolism and BBB permeability of morphine after long-term treatment, at both central and peripheral levels.Experimental approach: Male C57BL/6 mice were injected with morphine or saline solution for eight consecutive days in order to induce morphine analgesic tolerance. On the ninth day, both groups received a final injection of morphine (85%) and d3-morphine (morphine bearing three 2 H; 15%, w/w). Mice were then killed and blood, urine, brain and liver samples were collected. LC-MS/MS was used to quantify morphine, its metabolite morphine-3-glucuronide (M3G) and their respective d3-labelled forms.Key results: We found no significant differences in morphine CNS uptake and metabolism between control and tolerant mice. Interestingly, d3-morphine metabolism was decreased compared to morphine without any interference with our study.Conclusions and implications: Our data suggests that tolerance to the analgesic effects of morphine is not linked to increased glucuronidation to M3G or to altered global BBB permeability of morphine.
Published: 2018

28. A música é um jogo de criança

Author: François Delalande and François Delalande
Subjects: Music and children, Music--Instruction and study
Abstract: Este livro, publicado originalmente em 1984, em Paris, na França, traz importantes contribuições do pesquisador François Delalande acerca das relações entre a música e as crianças, ainda pouco exploradas no Brasil e em muitas partes do mundo. Por meio de dez diálogos radiofônicos com Jack Vidal e Guy Reibel, veiculados na Radio France, e depois editados pelo próprio autor, Delalande nos conduz a refletir sobre a música como um jogo de criança e entender a Educação Musical como um caminho para o despertar para a música, a descoberta da motivação, do desejo de fazer, escutar e produzir música. Esses diálogos revelam ideias e proposições poderosas, acerca da escuta, e dos modos de jogo na música, que ajudam a reinventar seus percursos nos territórios da Educação.
Published: 2019

29. Postscript: Music composition, creativity and collaboration – an old story, only recently told

Author: François Delalande
Subjects: Literature, business.industry, media_common.quotation_subject, Experimental and Cognitive Psychology, Musical composition, business, Psychology, Creativity, Music, Visual arts, media_common
Published: 2016

30. Post-Translational Regulation of the Glucose-6-Phosphatase Complex by Cyclic Adenosine Monophosphate Is a Crucial Determinant of Endogenous Glucose Production and Is Controlled by the Glucose-6-Phosphate Transporter

Author: Maud Soty, Gilles Mithieux, François Delalande, Carine Zitoun, Julien Chilloux, Amandine Gautier-Stein, Fabrice Bertile, Nutrition, diabète et cerveau, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Di Carlo, Marie-Ange, and Nutrition, diabète et cerveau (NUDICE)
Subjects: 0301 basic medicine, post-translational modification (PTM), Biochemistry, Antiporters, chemistry.chemical_compound, Cyclic AMP, Phosphorylation, ADCY6, glucose-6-phosphatase, Forskolin, Type 2 diabetes, Hep G2 Cells, Biological Sciences, ADCY3, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Glucose 6-phosphatase, Signal Transduction, Biochemistry & Molecular Biology, Monosaccharide Transport Proteins, Genetic Vectors, Glucose-6-Phosphate, Biology, Glucagon, Cell Line, 03 medical and health sciences, Animals, Humans, Cyclic adenosine monophosphate, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Protein kinase A, Adenoviruses, Human, glucose-6-phosphate transporter, Colforsin, Epithelial Cells, General Chemistry, Cyclic AMP-Dependent Protein Kinases, Rats, Protein Subunits, Glucose, gluconeogenesis, 030104 developmental biology, Gene Expression Regulation, glucagon, chemistry, Glucose 6-phosphate, Protein Biosynthesis, Chemical Sciences, Mutagenesis, Site-Directed, biology.protein, protein kinase A, Caco-2 Cells
Abstract: International audience; The excessive endogenous glucose production (EGP) induced by glucagon participates in the development of type 2 diabetes. To further understand this hormonal control, we studied the short-term regulation by cyclic adenosine monophosphate (cAMP) of the glucose-6-phosphatase (G6Pase) enzyme, which catalyzes the last reaction of EGP. In gluconeogenic cell models, a 1-h treatment by the adenylate cyclase activator forskolin increased G6Pase activity and glucose production independently of any change in enzyme protein amount or G6P content. Using specific inhibitors or protein overexpression, we showed that the stimulation of G6Pase activity involved the protein kinase A (PKA). Results of site-directed mutagenesis, mass spectrometry analyses, and in vitro phosphorylation experiments suggested that the PKA stimulation of G6Pase activity did not depend on a direct phosphorylation of the enzyme. However, the temperature-dependent induction of both G6Pase activity and glucose release suggested a membrane-based mechanism. G6Pase is composed of a G6P transporter (G6PT) and a catalytic unit (G6PC). Surprisingly, we demonstrated that the increase in G6PT activity was required for the stimulation of G6Pase activity by forskolin. Our data demonstrate the existence of a post-translational mechanism that regulates G6Pase activity and reveal the key role of G6PT in the hormonal regulation of G6Pase activity and of EGP.Erratum inCorrection to "Post-Translational Regulation of the Glucose-6-Phosphatase Complex by Cyclic Adenosine Monophosphate Is a Crucial Determinant of Endogenous Glucose Production and Is Controlled by the Glucose-6-Phosphate Transporter". [J Proteome Res. 2016]PMID: 27111186 DOI: 10.1021/acs.jproteome.6b00284
Published: 2016

31. RdCVF, A TRUNCATED THIOREDOXIN EVOLUTION LINKS REDOX HOMEOSTASIS TO GLUCOSE METABOLISM IN RETINA

Author: Najate Aït-Ali, Frédéric Blond, Francois Delalande, Alain Van Dorsselaer, and Thierry Léveillard
Subjects: Neurosciences. Biological psychiatry. Neuropsychiatry, RC321-571
Published: 2023
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32. Studying the fate of tumor extracellular vesicles at high spatio-temporal resolution using the zebrafish embryo

Author: Guillaume van Niel, Vincent Hyenne, Mayeul Collot, Pedro Machado, Frederik J. Verweij, Nina Fekonja, Andrey S. Klymchenko, Jacky G. Goetz, Joanna Bons, Christine Carapito, François Delalande, Jack Bauer, Susana Garcia Silva, Sébastien Harlepp, Olivier Lefebvre, Yannick Schwab, Ignacio Busnelli, Luc Mercier, Ana Amor López, Héctor Peinado, and Shima Ghoroghi
Subjects: Stromal cell, Animal model, In vivo, Tumor progression, media_common.quotation_subject, Blood circulation, Zebrafish embryo, Biology, Internalization, Extracellular vesicles, media_common, Cell biology
Abstract: SummaryTumor extracellular vesicles (tumor EVs) mediate the communication between tumor and stromal cells mostly to the benefit of tumor progression. Notably, tumor EVs have been reported to travel in the blood circulation, reach specific distant organs and locally modify the microenvironment. However, visualizing these eventsin vivostill faces major hurdles. Here, we show a new method for tracking individual circulating tumor EVs in a living organism: we combine novel, bright and specific fluorescent membrane probes, MemBright, with the transparent zebrafish embryo as an animal model. We provide the first description of tumor EVs’ hemodynamic behavior and document their arrest before internalization. Using transgenic lines, we show that circulating tumor EVs are uptaken by endothelial cells and blood patrolling macrophages, but not by leukocytes, and subsequently stored in acidic degradative compartments. Finally, we prove that the MemBright can be used to follow naturally released tumor EVsin vivo. Overall, our study demonstrates the usefulness and prospects of zebrafish embryo to track tumor EVsin vivo.HighlightsMemBright, a new family of membrane probes, allows for bright and specific staining of EVsZebrafish melanoma EVs are very similar to human and mouse melanoma EVs in morphology and protein contentThe zebrafish embryo is an adapted model to precisely track tumor EVs dynamics and fate in a living organism from light to electron microscopyCirculating tumor EVs are rapidly uptaken by endothelial cells and patrolling macrophagesCorrelated light and electron microscopy can be used in zebrafish to identify cells and compartments uptaking tumor EVsBlurbDispersion of tumor extracellular vesicles (EVs) throughout the body promotes tumor progression. However the behavior of tumor EVs in body fluids remains mysterious due to their small size and the absence of adapted animal model. Here we show that the zebrafish embryo can be used to track circulating tumor EVsin vivoand provide the first high-resolution description of their dissemination and uptake.
Published: 2018
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33. 5. Quelques conséquences de l’origine sensori-motrice de l’expérience musicale

Author: François Delalande
Published: 2018

34. The ESCRT-III Protein CHMP1A Mediates Secretion of Sonic Hedgehog on a Distinctive Subtype of Extracellular Vesicles

Author: Song Pang, Frank M. J. Jacobs, Michael E. Coulter, Raphael Gaudin, C. Shan Xu, Gerrald A Lodewijk, Ganeshwaran H. Mochida, Sarah Cianférani, Harald F. Hess, Wei-Chung Allen Lee, Christopher A. Walsh, Tomas Kirchhausen, Dilenny M. Gonzalez, Monica L. Calicchio, François Delalande, Edward Yang, Cristina M. Dorobantu, Richard S. Smith, Hart G.W. Lidov, David Haussler, Eric T. Wong, Vijay S. Ganesh, Maria K. Lehtinen, Elaine T. Lim, Thorsten M. Schlaeger, Molecular Neuroscience (SILS, FNWI), Equipe Direction scientifique, Sciences et Technologies de la Musique et du Son (STMS), Institut de Recherche et Coordination Acoustique/Musique (IRCAM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche et Coordination Acoustique/Musique (IRCAM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Computer Science and Artificial Intelligence Laboratory [Cambridge] (CSAIL), Massachusetts Institute of Technology (MIT), McMaster Univ, Med Phys & Appl Radiat Sci Dept, Hamilton, ON L8S 4K1, Canada, Analytic and Translational Genetics Unit, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Harvard Medical School [Boston] (HMS), Center for Biomolecular Science and Engineering, University of California [Santa Cruz] (UCSC), and University of California-University of California
Subjects: 0301 basic medicine, [SDV]Life Sciences [q-bio], Medical Physiology, Vesicular Transport Proteins, Aucun, Mice, 0302 clinical medicine, microcephaly, Sonic hedgehog, Pediatric, biology, neurodevelopment, Chemistry, Brain, Cell biology, Stem Cell Research - Nonembryonic - Non-Human, Adult, Endosome, 1.1 Normal biological development and functioning, Pontocerebellar hypoplasia, CHMP1A, Sciences du Vivant [q-bio]/Médecine humaine et pathologie, Article, General Biochemistry, Genetics and Molecular Biology, ESCRT, multivesicular body, 03 medical and health sciences, Extracellular Vesicles, sonic hedgehog, Rare Diseases, Clinical Research, Underpinning research, medicine, Animals, Humans, Secretion, Hedgehog Proteins, Progenitor, Endosomal Sorting Complexes Required for Transport, Infant, Newborn, Neurosciences, Infant, medicine.disease, Newborn, Stem Cell Research, Brain Disorders, 030104 developmental biology, Choroid Plexus, biology.protein, NIH 3T3 Cells, Congenital Structural Anomalies, Biochemistry and Cell Biology, 030217 neurology & neurosurgery, RAB18, Biogenesis
Abstract: SUMMARY Endosomal sorting complex required for transport (ESCRT) complex proteins regulate biogenesis and release of extracellular vesicles (EVs), which enable cell-to-cell communication in the nervous system essential for development and adult function. We recently showed human loss-of-function (LOF) mutations in ESCRT-III member CHMP1A cause autosomal recessive microcephaly with pontocerebellar hypoplasia, but its mechanism was unclear. Here, we show Chmp1a is required for progenitor proliferation in mouse cortex and cerebellum and progenitor maintenance in human cerebral organoids. In Chmp1a null mice, this defect is associated with impaired sonic hedgehog (Shh) secretion and intraluminal vesicle (ILV) formation in multivesicular bodies (MVBs). Furthermore, we show CHMP1A is important for release of an EV subtype that contains AXL, RAB18, and TMED10 (ART) and SHH. Our findings show CHMP1A loss impairs secretion of SHH on ART-EVs, providing molecular mechanistic insights into the role of ESCRT proteins and EVs in the brain., Graphical Abstract, In Brief Extracellular vesicles (EVs) are essential for cell-to-cell communication in developing brain. Coulter et al. show that the human microcephaly gene CHMP1A is required for neuroprogenitor proliferation through regulation of vesicular secretion of the growth factor sonic hedgehog (SHH). CHMP1A specifically impairs SHH secretion on a distinctive EV subtype, ART-EV.
Published: 2018

35. Validating Missing Proteins in Human Sperm Cells by Targeted Mass-Spectrometry- and Antibody-based Methods

Author: Yves Vandenbrouck, Charlotte Macron, Thomas Fréour, Lydie Lane, Karine Rondel, Charles Pineau, Paula D. Duek, François Delalande, Christine Carapito, Marine Seffals, Cecilia Lindskog, Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Swiss Institute of Bioinformatics [Genève] (SIB), H2P2 - Histo Pathologie Hight Precision (H2P2), Université de Rennes (UR)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Uppsala University, Centre hospitalier universitaire de Nantes (CHU Nantes), Centre de Recherche en Transplantation et Immunologie (U1064 Inserm - CRTI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Swiss Institute of Bioinformatics [Lausanne] (SIB), Université de Lausanne = University of Lausanne (UNIL), French National Agency for Research (ANR) [ANR-10-INBS-08], Biogenouest, Infrastructures en Biologie Sante et Agronomie (IBiSA), Conseil Regional de Bretagne, ANR-10-INBS-0008,ProFI,Infrastructure Française de Protéomique(2010), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Université de Lausanne (UNIL), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Foie, métabolismes et cancer, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Equipe Direction scientifique, Sciences et Technologies de la Musique et du Son (STMS), Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS), Etude de la dynamique des protéomes (EDyP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Département de science des protéines humaines [Genève], Université de Genève (UNIGE)-Faculté de médecine [Genève], Groupe d'Etude de la Reproduction Chez l'Homme et les Mammiferes (GERHM), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-IFR140-Institut National de la Santé et de la Recherche Médicale (INSERM), Jonchère, Laurent, and Infrastructure Française de Protéomique - - ProFI2010 - ANR-10-INBS-0008 - INBS - VALID
Subjects: 0301 basic medicine, Male, Proteome, [SDV]Life Sciences [q-bio], Tandem mass spectrometry, Biochemistry, immunocytochemistry, Tandem Mass Spectrometry, Testis, Human proteome project, ComputingMilieux_MISCELLANEOUS, ddc:616, biology, Histocytochemistry, bioinformatics, Immunohistochemistry, Spermatozoa, [SDV] Life Sciences [q-bio], spermatozoon, Chromosomes, Human, Pair 2, immunohistochemistry, Antibody, missing proteins, Human, human proteome project, targeted proteomics, Octoxynol, Human Protein Atlas, Context (language use), Computational biology, Chromosomes, Antibodies, 03 medical and health sciences, Spermatozoa/chemistry, Proteome/analysis, Humans, Chromosomes, Human, Pair 14, 030102 biochemistry & molecular biology, Testis/chemistry, General Chemistry, data mining, Sperm, Molecular biology, parallel reaction monitoring, 030104 developmental biology, Targeted mass spectrometry, biology.protein, Pair 14/genetics, Pair 2/genetics
Abstract: The present study is a contribution to the "neXt50 challenge", a coordinated effort across C-HPP teams to identify the 50 most tractable missing proteins (MPs) on each chromosome. We report the targeted search of 38 theoretically detectable MPs from chromosomes 2 and 14 in Triton X-100 soluble and insoluble sperm fractions from a total of 15 healthy donors. A targeted mass-spectrometry-based strategy consisting of the development of LC-PRM assays (with heavy labeled synthetic peptides) targeting 92 proteotypic peptides of the 38 selected MPs was used. Out of the 38 selected MPs, 12 were identified with two or more peptides and 3 with one peptide after extensive SDS-PAGE fractionation of the two samples and with overall low-intensity signals. The PRM data are available via ProteomeXchange in PASSEL (PASS01013). Further validation by immunohistochemistry on human testes sections and cytochemistry on sperm smears was performed for eight MPs with antibodies available from the Human Protein Atlas. Deep analysis of human sperm still allows the validation of MPs and therefore contributes to the C-HPP worldwide effort. We anticipate that our results will be of interest to the reproductive biology community because an in-depth analysis of these MPs may identify potential new candidates in the context of human idiopathic infertilities.
Published: 2017

36. FACTORS INFLUENCING REIMBURSEMENT OF MEDICAL DEVICES IN FRANCE

Author: Marie-Christine Reymond, Sylvia Germain, François Delalande, and Pierre Loge
Subjects: medicine.medical_specialty, Technology Assessment, Biomedical, Multivariate analysis, media_common.quotation_subject, Technical standard, 030209 endocrinology & metabolism, Reimbursement Mechanisms, 03 medical and health sciences, Government Agencies, 0302 clinical medicine, Humans, Medicine, Quality (business), 030212 general & internal medicine, Reimbursement, media_common, business.industry, Health Policy, Univariate, Evidence-based medicine, Clinical trial, Equipment and Supplies, Family medicine, France, business, Inclusion (education)
Abstract: Objectives: This study aims to analyze the key factors considered for the first application of the National Committee for the Evaluation of Medical Devices (CNEDiMTS) for achieving reimbursement through registration in the list of products and services qualifying for reimbursement (LPPR).Methods: All the appraisals studied on medical devices (MD) for first inclusion in the LPPR during 2011 and 2012 were retrieved from the French National Authority for Health or Haute Autorité de santé (HAS) Web site. A list of relevant factors was analyzed for each included opinion, followed by univariate and multivariate analyses to highlight the key factors that impacted the expected benefit (EB) provided by HAS.Results: A total of 151 appraisals were included in the study. Of them, 94 (62 percent) were granted with sufficient EB. The manufacturers were mostly from the United States (36 percent), while most of the applicants were from France (84 percent). After adjusting for other retrieved factors, it was observed that MDs complying with the technical standards, requests supported by opinion(s) from previous generation of MD, and the presence of recommendations or guidelines had more probability to obtain a sufficient EB. A lower probability was related to MDs supported by low-quality studies and with no specific health public benefit.Conclusions: Our results confirmed that manufacturers seeking reimbursement should be aware of the expectations of the health authorities (level of evidence, technical standard, etc.) and foresee their plan of sending requests for funding so that they can provide evidence of good quality.
Published: 2015

37. A ruthenium anticancer compound interacts with histones and impacts differently on epigenetic and death pathways compared to cisplatin

Author: Cynthia Licona, François Delalande, Moussa Ali, Marie-Elodie Spaety, Georg Mellitzer, John Spencer, Christian Gaiddon, Antonelle Capuozzo, Sarah Cianférani, Alain Van Dorsselaer, Rita Santamaria, Olivier Armant, Michel Pfeffer, Licona, Cynthia, Spaety, Marie Elodie, Capuozzo, Antonella, Ali, Moussa, Santamaria, Rita, Armant, Olivier, Delalande, Francoi, Van Dorsselaer, Alain, Cianferani, Sarah, Spencer, John, Pfeffer, Michel, Mellitzer, Georg, Gaiddon, Christian, INSERM 1113 Voies de signalisation du développement et du stress cellulaire dans les cancers digestifs et urologiques, 'Federico II' University of Naples Medical School, Institut de Chimie de Strasbourg, Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Karlsruhe Institute of Technology (KIT), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Department of Chemistry, School of Life Sciences, University of Sussex, and Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
Subjects: 0301 basic medicine, p53, Aucun, cisplatin, Sciences du Vivant [q-bio]/Cancer, Epigenesis, Genetic, Histones, 0302 clinical medicine, Neoplasms, Gene expression, Gene Regulatory Networks, biology, Epigenetic, Endoplasmic Reticulum Stress, Ruthenium, Gene Expression Regulation, Neoplastic, Histone, Oncology, Biochemistry, 030220 oncology & carcinogenesis, ER stre, Signal transduction, ER stress, Research Paper, QD0241, medicine.drug, Life sciences, Cell Survival, chemistry.chemical_element, 03 medical and health sciences, Histone H3, Cell Line, Tumor, ddc:570, Organometallic Compounds, medicine, Humans, [CHIM]Chemical Sciences, Epigenetics, ruthenium, Cell Proliferation, QD0415, Cisplatin, epigenetics, Gene Expression Profiling, HCT116 Cells, 030104 developmental biology, chemistry, biology.protein, Histone deacetylase, QD0146
Abstract: Ruthenium complexes are considered as potential replacements for platinum compounds in oncotherapy. Their clinical development is handicapped by a lack of consensus on their mode of action. In this study, we identify three histones (H3.1, H2A, H2B) as possible targets for an anticancer redox organoruthenium compound (RDC11). Using purified histones, we confirmed an interaction between the ruthenium complex and histones that impacted on histone complex formation. A comparative study of the ruthenium complex versus cisplatin showed differential epigenetic modifications on histone H3 that correlated with differential expression of histone deacetylase (HDAC) genes. We then characterized the impact of these epigenetic modifications on signaling pathways employing a transcriptomic approach. Clustering analyses showed gene expression signatures specific for cisplatin (42%) and for the ruthenium complex (30%). Signaling pathway analyses pointed to specificities distinguishing the ruthenium complex from cisplatin. For instance, cisplatin triggered preferentially p53 and folate biosynthesis while the ruthenium complex induced endoplasmic reticulum stress and trans-sulfuration pathways. To further understand the role of HDACs in these regulations, we used suberanilohydroxamic acid (SAHA) and showed that it synergized with cisplatin cytotoxicity while antagonizing the ruthenium complex activity. This study provides critical information for the characterization of signaling pathways differentiating both compounds, in particular, by the identification of a non-DNA direct target for an organoruthenium complex.
Published: 2017
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38. Endogenous morphine-6-glucuronide (M6G) is present in the plasma of patients: Validation of a specific anti-M6G antibody for clinical and basic research

Author: Marc Lamshöft, Alexis Laux-Biehlmann, François Delalande, Alain Van Dorsselaer, Hélène Chung, Jinane Mouheiche, Francis Schneider, Pierrick Poisbeau, Stéphanie Soldevila, Yannick Goumon, Julie Vérièpe, Patrick Garnero, Laurent Lamarque, Ingeborg Welters, and Herve Bazin
Subjects: biology, business.industry, Clinical Biochemistry, Endogeny, General Medicine, Morphine-6-glucuronide, Pharmacology, medicine.disease, Biochemistry, Sepsis, Polyclonal antibodies, Basic research, Dopamine, biology.protein, Morphine, Molecular Medicine, Medicine, Antibody, business, medicine.drug
Abstract: Endogenous morphine and its derivatives (morphine-6-glucuronide [M6G]; morphine-3-glucuronide [M3G]) are formed by mammalian cells from dopamine. Changes in the concentrations of endogenous morphine have been demonstrated in several pathologies (sepsis, Parkinson's disease, etc.), and they might be relevant as pathological markers. While endogenous morphine levels are detectable using enzyme-linked immunosorbant assay (ELISA), mass spectrometry (MS) analysis was, so far, the only approach to detect and quantify M6G. This study describes the preparation of a specific anti-M6G rabbit polyclonal antibody and its validation. The specificity of this antibody was assessed against 30 morphine-related compounds. Then, a M6G-specific ELISA-assay was tested to quantify M6G in the plasma of healthy donors, morphine-treated, and critically ill patients. The antibody raised against M6G displays a strong affinity for M6G, codeine-6-glucuronide, and morphine-3-6-glucuronide, whereas only weak cross-reactivities were observed for the other compounds. Both M6G-ELISA and LC-MS/MS approaches revealed the absence of M6G in the plasma of healthy donors (controls, n = 8). In all positive donors treated with morphine-patch (n = 5), M6G was detected using both M6G-ELISA and LC-MS/MS analysis. Finally, in a study on critically ill patients with circulating endogenous morphine (n = 26), LC-MS/MS analysis revealed that 73% of the positive-patients (19 of 26), corresponding to high M6G-levels in M6G-ELISA, contained M6G. In conclusion, we show that endogenous M6G can be found at higher levels than morphine in the blood of morphine-naive patients. With respect to the interest of measuring endogenous M6G in pathologies, we provide evidences that our ELISA procedure represents a powerful tool as it can easily and specifically detect endogenous M6G levels.
Published: 2013

39. Correction to 'Post-Translational Regulation of the Glucose-6-Phosphatase Complex by Cyclic Adenosine Monophosphate Is a Crucial Determinant of Endogenous Glucose Production and Is Controlled by the Glucose-6-Phosphate Transporter'

Author: Maud Soty, Julien Chilloux, François Delalande, Carine Zitoun, Fabrice Bertile, Gilles Mithieux, and Amandine Gautier-Stein
Subjects: General Chemistry, Biochemistry
Published: 2016

40. Maltaricin CPN, a new class IIa bacteriocin produced by Carnobacterium maltaromaticum CPN isolated from mould-ripened cheese

Author: Saïd Ennahar, Ikram Hammi, François Delalande, Rajae Belkhou, Sarah Cianférani, and Eric Marchioni
Subjects: 0301 basic medicine, 030106 microbiology, medicine.disease_cause, Gram-Positive Bacteria, Applied Microbiology and Biotechnology, Microbiology, 03 medical and health sciences, Bacteriocin, Listeria monocytogenes, Bacteriocins, Cheese, medicine, Animals, Amino Acid Sequence, chemistry.chemical_classification, biology, Strain (chemistry), Edman degradation, General Medicine, biology.organism_classification, Antimicrobial, Amino acid, Anti-Bacterial Agents, 030104 developmental biology, chemistry, Listeria, Carnobacterium, Bacteria, Biotechnology
Abstract: Aims The purpose of this study was to isolate, characterize and determine the structure and the antibacterial activities of a bacteriocin produced by Carnobacterium maltaromaticum CPN, a strain isolated from unpasteurized-milk Camembert cheese. Methods and Results This bacteriocin, termed maltaricin CPN, was produced at higher amounts in MRS broth at temperatures between 15 and 25°C. It was purified to homogeneity from culture supernatant by using a simple method consisting of cation-exchange and reversed-phase chromatographies. Mass spectrometry showed that maltaricin was a 4427.29 Da bacteriocin. Its amino acid sequence was determined by Edman degradation which showed that it had close similarity with bacteriocins of the class IIa. Maltaricin CPN consisted in fact of 44 unmodified amino acids including 2 cysteine residues at positions 9 and 14 linked by disulphide bond. The antimicrobial activity of maltaricin CPN covered a range of bacteria, with strong activity against many species of Gram-positive bacteria, especially the food-borne pathogen Listeria monocytogenes, but no activity against Gram-negative ones. Conclusions In the studied conditions, Carnobacterium maltaromaticum CPN produced a new class IIa bacteriocin with strong anti-Listeria activity. Significance and impact of the study The study covers the purification and the structural characterization of a new bacteriocin produced by strain Carnobacterium maltaromaticum CPN isolated from Camembert cheese. Its activity against strains of L. monocytogenes and higher production rates at relatively low temperatures show potential technological applications to improve the safety of refrigerated food. This article is protected by copyright. All rights reserved.
Published: 2016

41. Cdx2 homeoprotein inhibits non-homologous end joining in colon cancer but not in leukemia cells

Author: Isabelle Duluc, Benjamin Renouf, Jean-Noël Freund, François Delalande, Marie Vanier, Christine Soret, Thoueiba Saandi, Isabelle Gross, Claire Domon-Dell, and Elisabeth Martin
Subjects: DNA End-Joining Repair, Ku80, Transcription, Genetic, Cell Survival, DNA repair, DNA-Activated Protein Kinase, Genome Integrity, Repair and Replication, Biology, DNA-binding protein, DNA repair complex, Cell Line, Tumor, Genetics, Humans, CDX2 Transcription Factor, Protein Interaction Domains and Motifs, CDX2, Ku Autoantigen, Transcription factor, Etoposide, Homeodomain Proteins, Leukemia, Tumor Suppressor Proteins, Antigens, Nuclear, digestive system diseases, DNA-Binding Proteins, Non-homologous end joining, Colonic Neoplasms, embryonic structures, Cancer research, Ectopic expression
Abstract: Cdx2, a gene of the paraHox cluster, encodes a homeodomain transcription factor that plays numerous roles in embryonic development and in homeostasis of the adult intestine. Whereas Cdx2 exerts a tumor suppressor function in the gut, its abnormal ectopic expression in acute leukemia is associated to a pro-oncogenic function. To try to understand this duality, we have hypothesized that Cdx2 may interact with different protein partners in the two tissues and set up experiments to identify them by tandem affinity purification. We show here that Cdx2 interacts with the Ku heterodimer specifically in intestinal cells, but not in leukemia cells, via its homeodomain. Ku proteins do not affect Cdx2 transcriptional activity. However, Cdx2 inhibits in vivo and in vitro the DNA repair activity mediated by Ku proteins in intestinal cells. Whereas Cdx2 does not affect the recruitment of Ku proteins and DNA-PKcs into the DNA repair complex, it inhibits DNA-PKcs activity. Thus, we report here a new function of Cdx2, acting as an inhibitor of the DNA repair machinery, that may contribute to its tumor suppressor function specifically in the gut.
Published: 2011

42. Occurrence of Vibrio cholerae non-O1 in three wastewater treatment plants in Agadir (Morocco)

Author: Jean-Michel Scheftel, Olivier Meunier, Wardi Moussaoui, Alain Van Dorsselaer, Rkia Eddabra, Gilles Prévost, Rachida Mimouni, and François Delalande
Subjects: Gel electrophoresis, Nalidixic acid, biology, Physiology, Dendrogram, General Medicine, biology.organism_classification, medicine.disease_cause, Applied Microbiology and Biotechnology, Microbiology, Vibrio cholerae non-O1, Vibrionaceae, Vibrio cholerae, medicine, Pulsed-field gel electrophoresis, Typing, Biotechnology, medicine.drug
Abstract: A total of 21 isolates of Vibriocholerae non-O1 strains were isolated from three wastewater treatment plants in Agadir, Morocco. The isolates were analyzed by biochemical analysis, antibiogram, pulsed-field gel electrophoresis and the MALDI-TOF patterns of their protein masses were compared. Over 67% of isolates were susceptible to antimicrobial agents tested and 14% proved resistant to both trimethoprim-sulfamethoxazole and nalidixic acid. Typing by pulsed-field gel electrophoresis with NotI digestion revealed that the V. cholerae non-O1 strains from Agadir (Morocco) have a lower level of genetic homogeneity, the restriction patterns of whole-chromosomal DNA grouped the V. cholerae O1 and V. alginolyticus strains into a separate cluster from V. metschnikovii and V. cholerae non-O1 isolates. Furthermore, to gain additional analytical accuracy and reliability in the analysis we used dendrogram based on MALDI-TOF spectral patterns generated by the BioTyper 1.1™ software. All m/z signatures of all strains tested indicate that the mass spectral data contained sufficient information to distinguish between strains of V. cholerae.
Published: 2010

43. Sound explorations from the ages of 10 to 37 months: the ontogenesis of musical conducts

Author: Silvia Cornara and François Delalande
Subjects: Communication, geography, geography.geographical_feature_category, business.industry, media_common.quotation_subject, Musical, Music education, Linguistics, Motion (physics), Education, Surprise, Variation (linguistics), Repetition (music), Set (psychology), Psychology, business, Music, Sound (geography), media_common
Abstract: One of the forms of first musical conduct is the exploration of sound sources. When young children produce sounds with any object, these sounds may surprise them and so they make the sounds again – not exactly the same, but introducing some variation. A process of repetition with slight changes is set in motion which can be analysed, as did Piaget, as a circular reaction, but which can be seen, from a musical standpoint, as the development of a ‘sound discovery’ by repetition and variation. It is an elementary form of ‘musical idea’ which is a central moment in the process of invention in music. Studying these first musical conducts was the aim of a three-year research project. The chief condition of observation consisted in analysing the behaviour of one child left alone in a room exploring the sound possibilities of an instrument (a zither or a pair of cymbals). Fifty-five children from the ages of 10 to 37 months were filmed twice in this situation of solitary exploration. The videos were transcribed a...
Published: 2010

44. Cdc48 and Ufd3, new partners of the ubiquitin protease Ubp3, are required for ribophagy

Author: Catherine Dargemont, François Delalande, Mélanie Bonizec, Mickael M. Cohen, Batool Ossareh-Nazari, Svetlana Dokudovskaya, Christine Schaeffer, Alain Van Dorsselaer, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), and Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Proteasome Endopeptidase Complex, Saccharomyces cerevisiae Proteins, Recombinant Fusion Proteins, medicine.medical_treatment, Saccharomyces cerevisiae, Cell Cycle Proteins, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biochemistry, 03 medical and health sciences, 0302 clinical medicine, Ubiquitin, Valosin Containing Protein, Endopeptidases, Genetics, medicine, Cell Cycle Protein, Molecular Biology, ComputingMilieux_MISCELLANEOUS, Adaptor Proteins, Signal Transducing, 030304 developmental biology, Adenosine Triphosphatases, 0303 health sciences, Protease, biology, Scientific Reports, biology.organism_classification, Yeast, Ubiquitin ligase, Cell biology, Proteasome, biology.protein, Ribosomes, 030217 neurology & neurosurgery, Deubiquitination
Abstract: Ubiquitin-dependent processes can be antagonized by substrate-specific deubiquitination enzymes involved in many cellular functions. In this study, we show that the yeast Ubp3-Bre5 deubiquitination complex interacts with both the chaperone-like Cdc48, a major actor of the ubiquitin and proteasome system, and Ufd3, a ubiquitin-binding cofactor of Cdc48. We observed that these partners are required for the Ubp3-Bre5-dependent and starvation-induced selective degradation of yeast mature ribosomes, also called ribophagy. By contrast, proteasome-dependent degradation does not participate in this process. Our data favour the idea that these factors cooperate to recognize and deubiquitinate specific substrates of ribophagy before their vacuolar degradation.
Published: 2010

45. The technological era of ‘sound’: a challenge for musicology and a new range of social practices

Author: François Delalande
Subjects: Musical notation, Musicology, Contemporary classical music, Technological revolution, Computer science, Aesthetics, Musical, Amateur, Music, Musical analysis, Computer Science Applications, Musical form, Visual arts
Abstract: The ‘technological revolution’ that took place in music during the twentieth century is equivalent to the revolution that took place between the twelfth and fourteenth century, which transformed musical notation into applications of technology related to creation. This second revolution, as well as the first one, concerns not only musical form, but also the social organisation related to music. The aesthetic of sound is the key factor (in all the genres of contemporary music), which is a major challenge for musical analysis. Society is reorganising itself, favouring the appropriation and amateur practices within musical creation. Musical research institutions – and particularly the GRM – develop new forms of collaboration with their audience and contribute to the constitution of a ‘horizontal’ society, based on exchange, in frank opposition with the ‘vertical’ society, based on a reduced number of producers and a large amount of consumers.
Published: 2007

46. Differentiation between fresh and frozen–thawed sea bass (Dicentrarchus labrax) fillets using two-dimensional gel electrophoresis

Author: Guillaume Duflos, Sylvain Marlard, Valérie Lencel, Alain Van Dorsselaer, François Delalande, Alexandre Dehaut, Christine Carapito, Mylène Delosière, Thierry Grard, Pierrette Ethuin, Université du Littoral Côte d'Opale (ULCO), Qualité des Produits Animaux (QuaPA), Institut National de la Recherche Agronomique (INRA), Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Région Nord-Pas-de-Calais, and Ethuin , Pierrette
Subjects: Fish Proteins, Frozen–thawed, Biogenic amines, Parvalbumins, [SDV]Life Sciences [q-bio], Food spoilage, Ingénierie des aliments, Analytical Chemistry, spectrométrie de masse, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Freezing, Animals, Humans, Food engineering, Fresh, Electrophoresis, Gel, Two-Dimensional, filet de poisson, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, 14. Life underwater, Sea bass, électrophorèse, gel de polyacrylamide, Polyacrylamide gel electrophoresis, congélation, Chromatography, Two-dimensional gel electrophoresis, 2D-electrophoresis, biology, decongelation, Chemistry, sea bass (dicentrarchus labrax), frozen–thawed, fresh, biogenic amines, cod Gadus-morhua, protein, muscle, fish, storage, mass-spectrometry, proteome analysis, high-pressure, quality change, parvalbumin, Sea bass (Dicentrarchus labrax), General Medicine, biology.organism_classification, amine biogène, Isoelectric point, Seafood, Total volatile, Bass, Dicentrarchus, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Food Science
Abstract: International audience; This study aimed to identify a protein marker that can differentiate between fresh skinless and frozen–thawed sea bass (Dicentrarchus labrax) fillets using the two-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Distinct gel patterns, due to proteins with low molecular weight and low isoelectric points, distinguished fresh fillets from frozen–thawed ones. Frozen–thawed fillets showed two specific protein spots as early as the first day of the study. However, these spots were not observed in fresh fillets until at least 13 days of storage between 0 and 4 °C, fillets were judged, beyond this period, fish were unfit for human consumption as revealed by complementary studies on fish spoilage indicators namely total volatile basic nitrogen and biogenic amines. Mass spectrometry identified the specific proteins as parvalbumin isoforms. Parvalbumins may thus be useful markers of differentiation between fresh and frozen–thawed sea bass fillets.
Published: 2015

47. Rod-derived cone viability factor promotes cone survival by stimulating aerobic glycolysis

Author: Emmanuel Moyse, Emmanuelle Clérin, Géraldine Millet-Puel, Deniz Dalkara, José-Alain Sahel, Najate Aït-Ali, Frédéric Bouillaud, Anne Olivier-Bandini, Thierry Léveillard, Xavier Nicol, Alain Van Dorsselaer, Frédéric Blond, Ram Fridlich, Jacques Bellalou, Leah C. Byrne, François Delalande, Ludivine Perrocheau, Sacha Reichman, Céline Jaillard, Institut de la Vision, Centre National de la Recherche Scientifique ( CNRS ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO), Helen Wills Neuroscience Institute, University of California [Berkeley], Sanofi Aventis R&D, Production de Protéines Recombinantes (Plate-Forme) ( PRPF ), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Physiologie de la reproduction et des comportements [Nouzilly] ( PRC ), Institut National de la Recherche Agronomique ( INRA ) -Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique ( CNRS ), 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 ), Sanofi, UPMC, Inserm, CNRS, EC, FFB, and ANR, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Production de Protéines Recombinantes (Plate-Forme) (PRPF), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), 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), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], genetic structures, Glucose uptake, Mutation, Missense, Biology, Retinal Cone Photoreceptor Cells, General Biochemistry, Genetics and Molecular Biology, Retina, Mice, Thioredoxins, Retinitis pigmentosa, medicine, Animals, Humans, [ SDV.OT ] Life Sciences [q-bio]/Other [q-bio.OT], Eye Proteins, Glucose Transporter Type 1, Biochemistry, Genetics and Molecular Biology(all), Glucose transporter, medicine.disease, Alkaline Phosphatase, 3. Good health, Cell biology, Glucose, Biochemistry, Anaerobic glycolysis, biology.protein, Basigin, GLUT1, sense organs, Thioredoxin, Glycolysis, Retinitis Pigmentosa
Abstract: SummaryRod-derived cone viability factor (RdCVF) is an inactive thioredoxin secreted by rod photoreceptors that protects cones from degeneration. Because the secondary loss of cones in retinitis pigmentosa (RP) leads to blindness, the administration of RdCVF is a promising therapy for this untreatable neurodegenerative disease. Here, we investigated the mechanism underlying the protective role of RdCVF in RP. We show that RdCVF acts through binding to Basigin-1 (BSG1), a transmembrane protein expressed specifically by photoreceptors. BSG1 binds to the glucose transporter GLUT1, resulting in increased glucose entry into cones. Increased glucose promotes cone survival by stimulation of aerobic glycolysis. Moreover, a missense mutation of RdCVF results in its inability to bind to BSG1, stimulate glucose uptake, and prevent secondary cone death in a model of RP. Our data uncover an entirely novel mechanism of neuroprotection through the stimulation of glucose metabolism.
Published: 2015

48. Proteome Analysis of Plant-Virus Interactome

Author: Christine Carapito, Jean Paul Brizard, Christophe Brugidou, Alain Van Dorsselaer, and François Delalande
Subjects: biology, Rice yellow mottle virus, Computational biology, biology.organism_classification, Biochemistry, Interactome, Virus, Analytical Chemistry, Cell biology, Viral replication, Plant virus, Proteome, Human proteome project, Protein biosynthesis, Molecular Biology
Abstract: Known host-parasite molecular interactions are widespread among parasite families, but these interactions have to be particularly large considering that viruses generally encode few proteins. Although some particular virus-host interactions are well described, no global study has yet shown multiple and simultaneous interactions in a host-parasite biological system. To prove that these multiple interactions occur in biological conditions, the complexes formed by a plant virus (rice yellow mottle virus) and the proteins of its natural host (rice) were extracted and purified from infected tissue sample. Remarkably mass spectrometry permitted the identification of a large number of proteins from the complexes that are involved in different functions not encoded by the virus but probably essential for its biological life cycle. This recruiting of proteins was strongly confirmed by the repetition of experiments using different pairs of virus-host and the use of high salt concentration to extract the complexes. We mainly identified proteins involved in plant defense, metabolism, translation, and protein synthesis and some proteins involved in transport. This study demonstrates that viruses are able to recruit many proteins from their hosts to ensure their development. Among different pairs of virus-host, similar protein functions were identified suggesting a particular importance of these proteins for viruses. The identification of particular paralog proteins among multigenic families suggests the high specificity of the recruiting for some protein functions.
Published: 2006

49. Diversity of the exoproteome of Fusarium graminearum grown on plant cell wall

Author: Emmanuelle Leize-Wagner, Florence Goubet, Vincent Phalip, François Delalande, Didier Hatsch, Christine Carapito, Paul Dupree, Jean-Marc Jeltsch, Alain Van Dorsselaer, Klotz, Evelyne, Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Substances naturelles/chimie moléculaire, Université Louis Pasteur - Strasbourg I-Ecole européenne de chimie, polymères et matériaux [Strasbourg]-Centre National de la Recherche Scientifique (CNRS), and Institut Pluridisciplinaire Hubert Curien (IPHC)
Subjects: Polysaccharides/chemistry, Humulus lupulus, MESH: Plants, MESH: Cellulose 1,4-beta-Cellobiosidase, Proteomics, Cellulose 1, Plant Proteins/*chemistry/metabolism, Fusarium, Cell Wall, Gene Expression Regulation, Plant, Glucose/metabolism, Non-U.S. Gov't, Plant Proteins, chemistry.chemical_classification, MESH: Fusarium, 0303 health sciences, MESH: Plant Proteins, biology, food and beverages, General Medicine, Plants, 4-beta-Cellobiosidase/chemistry/metabolism, MESH: Glucose, Biochemistry, Fusarium/*chemistry/growth & development, Fungus, Research Support, Microbiology, Cell wall, 03 medical and health sciences, MESH: Cell Wall, Polysaccharides, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Cellulose 1,4-beta-Cellobiosidase, Genetics, Extracellular, Secretion, MESH: Gene Expression Regulation, Plant, 030304 developmental biology, Plants/*microbiology, 030306 microbiology, Cell Wall/chemistry/*metabolism, Plant, biology.organism_classification, MESH: Polysaccharides, Glucose, Enzyme, Gene Expression Regulation, chemistry
Abstract: 0172-8083 (Print) Journal Article; The exoproteome of the fungus Fusarium graminearum grown on glucose and on hop (Humulus lupulus, L.) cell wall has been investigated. The culture medium was found to contain a higher quantity of proteins and the proteins are more diverse when the fungus is grown on cell wall. Using both 1D and 2D electrophoresis followed by mass spectrometry analysis and protein identification based on similarity searches, 84 unique proteins were identified in the cell wall-grown fungal exoproteome. Many are putatively implicated in carbohydrate metabolism, mainly in cell wall polysaccharide degradation. The predicted carbohydrate-active enzymes fell into 24 different enzymes classes, and up to eight different proteins within a same class are secreted. This indicates that fungal metabolism becomes oriented towards synthesis and secretion of a whole arsenal of enzymes able to digest almost the complete plant cell wall. Cellobiohydrolase is one of the only four proteins found both after growth on glucose and on plant cell wall and we propose that this enzyme could act as a sensor of the extracellular environment. Extensive knowledge of this very diverse F. graminearum exoproteome is an important step towards the full understanding of Fusarium/plants interactions.
Published: 2005

50. Multigenic families and proteomics: Extended protein characterization as a tool for paralog gene identification

Author: Christophe Brugidou, François Delalande, Jean-Paul Brizard, Alain Van Dorsselaer, and Christine Carapito
Subjects: Proteomics, Sequence analysis, Gene Expression, Biology, Genes, Plant, Biochemistry, Genome, Chromosomes, Plant, Mass Spectrometry, Homology (biology), Plant Viruses, Mitochondrial Proteins, Sequence Analysis, Protein, RNA interference, Fructose-Bisphosphate Aldolase, Databases, Genetic, Gene expression, Nanotechnology, Gene silencing, Gene Silencing, Databases, Protein, Molecular Biology, Gene, Phenylalanine Ammonia-Lyase, Plant Proteins, Genetics, fungi, Discriminant Analysis, Reproducibility of Results, Oryza, Chaperonin 60, Freeze Drying, Multigene Family, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, RNA Interference, Protein Processing, Post-Translational, Chromatography, Liquid
Abstract: In classical proteomic studies, the searches in protein databases lead mostly to the identification of protein functions by homology due to the non-exhaustiveness of the protein databases. The quality of the identification depends on the studied organism, its complexity and its representation in the protein databases. Nevertheless, this basic function identification is insufficient for certain applications namely for the development of RNA-based gene-silencing strategies, commonly termed RNA interference (RNAi) in animals and post-transcriptional gene silencing (PTGS) in plants, that require an unambiguous identification of the targeted gene sequence. A PTGS strategy was considered in the study of the infection of Oryza sativa by the Rice Yellow Mottle Virus (RYMV). It is suspected that the RYMV recruits host proteins after its entry into plant cells to form a complex facilitating virus multiplication and spreading. The protein partners of this complex were identified by a classical proteomic approach, nano liquid chromatography tandem mass spectrometry. Among the identified proteins, several were retained for a PTGS strategy. Nevertheless most of the protein candidates appear to be members of multigenic families for which all paralog genes are not present in protein databases. Thus the identification of the real expressed paralog gene with classical protein database searches is impossible. Consequently, as the genome contains all genes and thus all paralog genes, a whole genome search strategy was developed to determine the specific expressed paralog gene. With this approach, the identification of peptides matching only a single gene, called discriminant peptides, allows definitive proof of the expression of this identified gene. This strategy has several requirements: (i) a genome completely sequenced and accessible; (ii) high protein sequence coverage. In the present work, through three examples, we report and validate for the first time a genome database search strategy to specifically identify paralog genes belonging to multigenic families expressed under specific conditions.
Published: 2005

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