ABSTRACTSepticemia-causingVibrio vulnificusproduces at least three exoproteases, VvpE, VvpS, and VvpM, all of which participate in interactions with human cells. Expression of VvpE and VvpS is induced in the stationary phase by multiple transcription factors, including sigma factor S, SmcR, and the cAMP-cAMP receptor protein (cAMP-CRP) complex. Distinct roles of VvpM, such as induction of apoptosis, lead us to hypothesize VvpM expression is different from that of the other exoproteases. Its transcription, which was found to be independent of sigma S, is induced at the early exponential phase and then becomes negligible upon entry into the stationary phase. SmcR and CRP were studied regarding the control ofvvpMexpression. Transcription ofvvpMwas repressed by SmcR and cAMP-CRP complex individually, which specifically bound to the regions −2 to +20 and +6 to +27, respectively, relative to thevvpMtranscription initiation site. Derepression ofvvpMgene expression was 10- to 40-fold greater in ansmcR crpdouble mutant than in single-gene mutants. Therefore, these results show that the expression ofV. vulnificusexoproteases is differentially regulated, and in this way, distinct proteases can engage in specific interactions with a host.IMPORTANCEAn opportunistic human pathogen,Vibrio vulnificusproduces multiple extracellular proteases that are involved in diverse interactions with a host. The total exoproteolytic activity is detected mainly in the supernatants of the high-cell-density cultures. However, some proteolytic activity derived from a metalloprotease, VvpM, was present in the supernatants of the low-cell-density cultures sampled at the early growth period. In this study, we present the regulatory mechanism for VvpM expression via repression by at least two transcription factors. This type of transcriptional regulation is the exact opposite of those for expression of the otherV. vulnificusexoproteases. Differential regulation of each exoprotease's production then facilitates the pathogen's participation in the distinct interactions with a host.
Vibrio vulnificus rpoSExpression Is Repressed by Direct Binding of cAMP-cAMP Receptor Protein Complex to Its Two Promoter Regions