title: Quorum Sensing Determines the Choice of Antiphage Defense Strategy in Vibrio anguillarum.
abstract: Selection for phage resistance is a key driver of bacterial diversity and evolution, and phage-host interactions may therefore have strong influence on the genetic and functional dynamics of bacterial communities. In this study, we found that an important, but so far largely overlooked, determinant of the outcome of phage-bacterial encounters in the fish pathogen Vibrio anguillarum is bacterial cell-cell communication, known as quorum sensing. Specifically, V. anguillarum PF430-3 cells locked in the low-cell-density state (ΔvanT mutant) express high levels of the phage receptor OmpK, resulting in a high susceptibility to phage KVP40, but achieve protection from infection by enhanced biofilm formation. By contrast, cells locked in the high-cell-density state (ΔvanΟ mutant) are almost completely unsusceptible due to quorum-sensing-mediated downregulation of OmpK expression. The phenotypes of the two quorum-sensing mutant strains are accurately reflected in the behavior of wild-type V. anguillarum, which (i) displays increased OmpK expression in aggregated cells compared to free-living variants in the same culture, (ii) displays a clear inverse correlation between ompK mRNA levels and the concentration of N-acylhomoserine lactone quorum-sensing signals in the culture medium, and (iii) survives mainly by one of these two defense mechanisms, rather than by genetic mutation to phage resistance. Taken together, our results demonstrate that V. anguillarum employs quorum-sensing information to choose between two complementary antiphage defense strategies. Further, the prevalence of nonmutational defense mechanisms in strain PF430-3 suggests highly flexible adaptations to KVP40 phage infection pressure, possibly allowing the long-term coexistence of phage and host.
  
  
  OBJECTIVE
  Comprehensive knowledge on bacterial antiphage strategies and their regulation is essential for understanding the role of phages as drivers of bacterial evolution and diversity. In an applied context, development of successful phage-based control of bacterial pathogens also requires detailed understanding of the mechanisms of phage protection in pathogenic bacteria. Here, we demonstrate for the first time the presence of quorum-sensing-regulated phage defense mechanisms in the fish pathogen Vibrio anguillarum and provide evidence that quorum-sensing regulation allows V. anguillarum to alternate between different phage protection mechanisms depending on population cell density. Further, our results demonstrate the prevalence of nonmutational defense mechanisms in the investigated V. anguillarum strain, which allow flexible adaptations to a dynamic phage infection pressure.
authors.affiliation: Marine Biological Section, Department of Biology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Demeng
authors.initials: D
authors.lastname: Tan
authors.affiliation: Section for Biomolecular Sciences, Department of Biology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Sine Lo
authors.initials: SL
authors.lastname: Svenningsen
authors.affiliation: Marine Biological Section, Department of Biology, University of Copenhagen, Copenhagen, Denmark mmiddelboe@bio.ku.dk.
authors.forename: Mathias
authors.initials: M
authors.lastname: Middelboe
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001435:Bacteriophages / Q000254:growth & development*; D015964:Gene Expression Regulation, Bacterial; D006790:Host-Parasite Interactions; D053038:Quorum Sensing; D011991:Receptors, Virus / Q000378:metabolism; D014733:Vibrio / Q000502:physiology* / Q000821:virology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D011991:Receptors, Virus