Metalloprotease Vsm Is the Major Determinant of Toxicity for Extracellular Products of Vibrio splendidus
| Autor: | Denis Saulnier, Didier Mazel, Hélène Munier-Lehmann, Monique Zagorec, Claude Delsert, Marie-Christine Champomier-Vergès, Frédérique Le Roux, Johan Binesse |
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| Přispěvatelé: | Biologie systémique - Systems Biology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Génétique et Pathologie des Mollusques Marins, 17390 La Tremblade, France. (LGPMM), Santé, Génétique et Microbiologie des Mollusques (IFREMER SG2M), Institut Français de Recherche pour l'Exploitation de la Mer - Atlantique (IFREMER Atlantique), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer - Atlantique (IFREMER Atlantique), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Centre de recherche en Biologie Cellulaire (CRBM), Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1), Unité de recherche Flore Lactique et Environnement Carné (UFLEC), Institut National de la Recherche Agronomique (INRA), Institut Pasteur [Paris], Plasticité du Génome Bactérien (PGB), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Génétique et Pathologie (LGP), Amélioration génétique, du contrôle des performances et de la santé des mollusques marins (AGSAE), Chimie Organique, This study was carried out with financial assistance from the Institut Pasteur, the Centre National de la Recherche Scientifique (CNRS-URA 2171), the Institut Françcais de Recherche pour l'Exploitation de la Mer (IFREMER), and the Institut de Génomique Marine (Ministère de la Recherche contract no. 0425). Johan Binesse acknowledges a Ph.D. scholarship from the French Ministry of Research and Technology (MRT)., Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
MESH: Sequence Analysis
DNA Proteome MESH: Sequence Homology Amino Acid [SDV]Life Sciences [q-bio] Mutant species Applied Microbiology and Biotechnology Virulence factor Mice MESH: Animals Pathogen MESH: Bacterial Proteins Cells Cultured 0303 health sciences Metalloproteinase Ecology MESH: Genomics Genomics musculoskeletal system MESH: Mollusca MESH: Proteome cardiovascular system MESH: Metalloproteases Biotechnology MESH: Cells Cultured DNA Bacterial Virulence Factors education Molecular Sequence Data Bacillus thuringiensis Virulence Biology Microbiology 03 medical and health sciences MESH: Vibrio Bacterial Proteins Extracellular Invertebrate Microbiology Animals MESH: Mice 030304 developmental biology MESH: Virulence Factors Vibrio MESH: Bacillus thuringiensis MESH: Molecular Sequence Data Sequence Homology Amino Acid 030306 microbiology Wild type Sequence Analysis DNA Fibroblasts biology.organism_classification Molecular biology MESH: DNA Bacterial proteins MESH: Fibroblasts MESH: Gene Deletion Mollusca Metalloproteases Genomic Gene Deletion Food Science |
| Zdroj: | Applied and Environmental Microbiology Applied and Environmental Microbiology, American Society for Microbiology, 2008, 74 (23), pp.7108-7117. ⟨10.1128/AEM.01261-08⟩ Applied and Environmental Microbiology, 2008, 74 (23), pp.7108-7117. ⟨10.1128/AEM.01261-08⟩ Applied and environmental microbiology (0099-2240) (American society for microbiology), 2008-12, Vol. 74, N. 23, P. 7108-7117 |
| ISSN: | 0099-2240 1098-5336 |
| DOI: | 10.1128/AEM.01261-08⟩ |
| Popis: | Genomic data combined with reverse genetic approaches have contributed to the characterization of major virulence factors of Vibrio species; however, these studies have targeted primarily human pathogens. Here, we investigate virulence factors in the oyster pathogen Vibrio splendidus LGP32 and show that toxicity is correlated to the presence of a metalloprotease and its corresponding vsm gene. Comparative genomics showed that an avirulent strain closely related to LGP32 lacked the metalloprotease. The toxicity of LGP32 metalloprotease was confirmed by exposing mollusk and mouse fibroblastic cell lines to extracellular products (ECPs) of the wild type (wt) and a vsm deletion mutant (Δ vsm mutant). The ECPs of the wt induced a strong cytopathic effect whose severity was cell type dependent, while those of the Δ vsm mutant were much less toxic, and exposure to purified protein demonstrated the direct toxicity of the Vsm metalloprotease. Finally, to investigate Vsm molecular targets, a proteomic analysis of the ECPs of both LGP32 and the Δ vsm mutant was performed, revealing a number of differentially expressed and/or processed proteins. One of these, the VSA1062 metalloprotease, was found to have significant identity to the immune inhibitor A precursor, a virulence factor of Bacillus thuringiensis . Deletion mutants corresponding to several of the major proteins were constructed by allelic exchange, and the ECPs of these mutants proved to be toxic to both cell cultures and animals. Taken together, these data demonstrate that Vsm is the major toxicity factor in the ECPs of V. splendidus . |
| Databáze: | OpenAIRE |
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