title: The major outer membrane protein OmpU of Vibrio splendidus contributes to host antimicrobial peptide resistance and is required for virulence in the oyster Crassostrea gigas.
abstract: Vibrio splendidus, strain LGP32, is an oyster pathogen associated with the summer mortalities affecting the production of Crassostrea gigas oysters worldwide. Vibrio splendidus LGP32 was shown to resist to up to 10 microM Cg-Def defensin and Cg-BPI bactericidal permeability increasing protein, two antimicrobial peptides/proteins (AMPs) involved in C. gigas immunity. The resistance to both oyster Cg-Def and Cg-BPI and standard AMPs (polymyxin B, protegrin, human BPI) was dependent on the ompU gene. Indeed, upon ompU inactivation, minimal bactericidal concentrations decreased by up to fourfold. AMP resistance was restored upon ectopic expression of ompU. The susceptibility of bacterial membranes to AMP-induced damages was independent of the ompU-mediated AMP resistance. Besides its role in AMP resistance, ompU proved to be essential for the adherence of V. splendidus LGP32 to fibronectin. Interestingly, in vivo, ompU was identified as a major determinant of V. splendidus pathogenicity in oyster experimental infections. Indeed, the V. splendidus-induced oyster mortalities dropped from 56% to 11% upon ompU mutation (Kaplan-Meier survival curves, P < 0.01). Moreover, in co-infection assays, the ompU mutant was out competed by the wild-type strain with competitive indexes in the range of 0.1-0.2. From this study, ompU is required for virulence of V. splendidus. Contributing to AMP resistance, conferring adhesive properties to V. splendidus, and being essential for in vivo fitness, the OmpU porin appears as an essential effector of the C. gigas/V. splendidus interaction.
authors.affiliation: CNRS, UMR 5119, Laboratoire Ecosystèmes lagunaires, Université Montpellier 2, 34095 Montpellier, France.
authors.forename: Marylise
authors.initials: M
authors.lastname: Duperthuy
authors.forename: Johan
authors.initials: J
authors.lastname: Binesse
authors.forename: Frédérique
authors.initials: F
authors.lastname: Le Roux
authors.forename: Bernard
authors.initials: B
authors.lastname: Romestand
authors.forename: Audrey
authors.initials: A
authors.lastname: Caro
authors.forename: Patrice
authors.initials: P
authors.lastname: Got
authors.forename: Alain
authors.initials: A
authors.lastname: Givaudan
authors.forename: Didier
authors.initials: D
authors.lastname: Mazel
authors.forename: Evelyne
authors.initials: E
authors.lastname: Bachère
authors.forename: Delphine
authors.initials: D
authors.lastname: Destoumieux-Garzón
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000276:immunology; D001798:Blood Proteins / Q000276:immunology; D049896:Crassostrea / Q000276:immunology / Q000382:microbiology*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D009154:Mutation; D014733:Vibrio / Q000235:genetics / Q000472:pathogenicity*; D014735:Vibrio Infections / Q000382:microbiology*; D014774:Virulence
chemical_list: D018829:Adhesins, Bacterial; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; C096349:OmpU protein, Vibrio cholerae; C021623:bactericidal permeability increasing protein