title: Screening for inhibition of Vibrio cholerae VipA-VipB interaction identifies small-molecule compounds active against type VI secretion.
abstract: The type VI secretion system (T6SS) is the most prevalent bacterial secretion system and an important virulence mechanism utilized by Gram-negative bacteria, either to target eukaryotic cells or to combat other microbes. The components show much variability, but some appear essential for the function, and two homologues, denoted VipA and VipB in Vibrio cholerae, have been identified in all T6SSs described so far. Secretion is dependent on binding of an α-helical region of VipA to VipB, and in the absence of this binding, both components are degraded within minutes and secretion is ceased. The aim of the study was to investigate if this interaction could be blocked, and we hypothesized that such inhibition would lead to abrogation of T6S. A library of 9,600 small-molecule compounds was screened for their ability to block the binding of VipA-VipB in a bacterial two-hybrid system (B2H). After excluding compounds that showed cytotoxicity toward eukaryotic cells, that inhibited growth of Vibrio, or that inhibited an unrelated B2H interaction, 34 compounds were further investigated for effects on the T6SS-dependent secretion of hemolysin-coregulated protein (Hcp) or of phospholipase A1 activity. Two compounds, KS100 and KS200, showed intermediate or strong effects in both assays. Analogues were obtained, and compounds with potent inhibitory effects in the assays and desirable physicochemical properties as predicted by in silico analysis were identified. Since the compounds specifically target a virulence mechanism without affecting bacterial replication, they have the potential to mitigate the virulence with minimal risk for development of resistance.
authors.affiliation: Department of Clinical Microbiology, Clinical Bacteriology and Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: Kun
authors.initials: K
authors.lastname: Sun
authors.affiliation: Department of Clinical Microbiology, Clinical Bacteriology and Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: Jeanette
authors.initials: J
authors.lastname: Bröms
authors.affiliation: Department of Clinical Microbiology, Clinical Bacteriology and Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: Moa
authors.initials: M
authors.lastname: Lavander
authors.affiliation: Department of Clinical Microbiology, Clinical Bacteriology and Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: Bharat Kumar
authors.initials: BK
authors.lastname: Gurram
authors.affiliation: Department of Chemistry and Laboratories for Chemical Biology Umeå (LCBU), Umeå University, Umeå, Sweden.
authors.forename: Per-Anders
authors.initials: PA
authors.lastname: Enquist
authors.affiliation: Department of Chemistry and Laboratories for Chemical Biology Umeå (LCBU), Umeå University, Umeå, Sweden.
authors.forename: C David
authors.initials: CD
authors.lastname: Andersson
authors.affiliation: Department of Chemistry and Laboratories for Chemical Biology Umeå (LCBU), Umeå University, Umeå, Sweden.
authors.forename: Mikael
authors.initials: M
authors.lastname: Elofsson
authors.affiliation: Department of Clinical Microbiology, Clinical Bacteriology and Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden Anders.Sjöstedt@climi.umu.se.
authors.forename: Anders
authors.initials: A
authors.lastname: Sjöstedt
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000378:metabolism*; D058947:Bacterial Secretion Systems / Q000187:drug effects*; D006460:Hemolysin Proteins / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D054466:Phospholipases A1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014734:Vibrio cholerae / Q000187:drug effects* / Q000472:pathogenicity; D037521:Virulence Factors / Q000037:antagonists & inhibitors
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; C097322:Hcp protein, Vibrio cholerae; D006460:Hemolysin Proteins; D037521:Virulence Factors; D054466:Phospholipases A1