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Vibrio cholerae is an intestinal pathogen that causes the diarrheal disease cholera. Colonization of the intestine depends upon the expression of genes that allowV. cholerae to overcome host barriers, including low pH, bile acids, and the innate immune system. ToxR is a major contributor to this process. ToxR is a membrane-spanning transcription factor that coordinates gene expression in response to environmental cues. In previous work we showed that ToxR upregulatedleuO expression in response to bile salts. LeuO is a LysR family transcription factor that contributes to acid tolerance, bile resistance, and biofilm formation inV. cholerae . Here, we investigated the function of ToxR and LeuO in cationic antimicrobial peptide (CAMP) resistance. We report that ToxR and LeuO contribute to CAMP resistance by regulatingcarRS transcription. CarRS is a two-component regulatory system that positively regulatesalmEFG expression. AlmEFG confers CAMP resistance by glycinylation of lipid A. We found that the expression ofcarRS andalmEFG and the polymyxin B MIC increased in mutants lackingtoxRS orleuO . Conversely,leuO overexpression decreased the polymyxin B MIC. Furthermore, we found that LeuO directly bound to thecarRS promoter and that ToxR-dependent activation ofleuO transcription regulatedcarRS transcription in response to bile salts. Our results suggest that LeuO functions downstream of ToxR to modulatecarRS expression in response to environmental cues. This study extends the functional role of ToxR and LeuO in environmental adaptation to include cell surface remodeling and CAMP resistance.

Vibrio cholerae LeuO Links the ToxR Regulon to Expression of Lipid A Remodeling Genes