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In many bacteria, including  Vibrio cholerae , cyclic dimeric guanosine monophosphate (c-di-GMP) controls the motile to biofilm life style switch. Yet, little is known about how this occurs. In this study, we report that changes in c-di-GMP concentration impact the biosynthesis of the MshA pili, resulting in altered motility and biofilm phenotypes in  V .  cholerae . Previously, we reported that  cdgJ  encodes a c-di-GMP phosphodiesterase and a Δ cdgJ  mutant has reduced motility and enhanced biofilm formation. Here we show that loss of the genes required for the mannose - sensitive hemagglutinin  ( MshA )  pilus biogenesis restores motility in the Δ cdgJ  mutant. Mutations of the predicted ATPase proteins  mshE  or  pilT , responsible for polymerizing and depolymerizing MshA pili, impair near surface motility behavior and initial surface attachment dynamics. A Δ cdgJ  mutant has enhanced surface attachment, while the Δ cdgJmshA  mutant phenocopies the high motility and low attachment phenotypes observed in a Δ mshA  strain. Elevated concentrations of c-di-GMP enhance surface MshA pilus production. MshE, but not PilT binds c-di-GMP directly, establishing a mechanism for c-di-GMP signaling input in MshA pilus production. Collectively, our results suggest that the dynamic nature of the MshA pilus established by the assembly and disassembly of pilin subunits is essential for transition from the motile to sessile lifestyle and that c-di-GMP affects MshA pilus assembly and function through direct interactions with the MshE ATPase.

C-di-GMP Regulates Motile to Sessile Transition by Modulating MshA Pili Biogenesis and Near-Surface Motility Behavior in Vibrio cholerae