The Cyclic AMP Receptor Protein Modulates Colonial Morphology in Vibrio cholerae

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Abstract
Inactivation of the quorum-sensing regulator HapR causes Vibrio cholerae El Tor biotype strain C7258 to adopt a rugose colonial morphology that correlates with enhanced biofilm formation. V. cholerae mutants lacking the cyclic AMP (cAMP) receptor protein (CRP) produce very little HapR, which results in elevated expression of Vibrio exopolysaccharide ( vps ) genes and biofilm compared to the wild type. However, Δ crp mutants still exhibited smooth colonial morphology and expressed reduced levels of vps genes compared to isogenic hapR mutants. In this study we demonstrate that deletion of crp and cya (adenylate cyclase) converts a rugose Δ hapR mutant to a smooth one. The smooth Δ hapR Δ crp and Δ hapR Δ cya double mutants could be converted back to rugose by complementation with crp and cya , respectively. CRP was found to enhance the expression of VpsR, a strong activator of vps expression, but to diminish transcription of VpsT. Ectopic expression of VpsR in smooth Δ hapR Δ crp and Δ hapR Δ cya double mutants restored rugose colonial morphology. Lowering intracellular cAMP levels in a Δ hapR mutant by the addition of glucose diminished VpsR expression and colonial rugosity. On the basis of our results, we propose a model for the regulatory input of CRP on exopolysaccharide biosynthesis.