The Polar Flagellar Transcriptional Regulatory Network in Deviates from Canonical Species.

Swimming motility is a critical virulence factor in pathogenesis for numerous species. Vibrio campbellii DS40M4 is a wild-type isolate that has been recently established as a highly tractable model strain for bacterial genetics studies. We sought to exploit the tractability and relevance of this strain for characterization of flagellar gene regulation in . Using comparative genomics, we identified homologs of flagellar and chemotaxis genes conserved in other members of the and determined the transcriptional profile of these loci using differential RNA-seq. We systematically deleted all 63 predicted flagellar and chemotaxis genes in and examined their effects on motility and flagellum production. We specifically focused on the core regulators of the flagellar hierarchy established in other vibrios: RpoN (σ), FlrA, FlrC, and FliA. Our results show that transcription of flagellar and chemotaxis genes is governed by a multitiered regulatory hierarchy similar to other motile species. However, there are several critical differences in : (i) the σ-dependent regulator FlrA is dispensable for motility; (ii) the , , , and operons do not require σ for expression; and (iii) FlrA and FlrC coregulate class II genes. Our model proposes that the flagellar transcriptional hierarchy has three classes of genes, in contrast to the four-class hierarchy in Vibrio cholerae. Our genetic and phenotypic dissection of the flagellar regulatory network highlights the differences that have evolved in flagellar regulation across the . Vibrio campbellii is a Gram-negative bacterium that is free-living and ubiquitous in marine environments and is an important global pathogen of fish and shellfish. Disruption of the flagellar motor significantly decreases host mortality of , suggesting that motility is a key factor in pathogenesis. Using this model organism, we identified >60 genes that encode proteins with predicted structural, mechanical, or regulatory roles in function of the single polar flagellum in . We systematically tested strains containing single deletions of each gene to determine the impact on motility and flagellum production. Our studies have uncovered differences in the regulatory network and function of several genes in compared to established systems in Vibrio cholerae and Vibrio parahaemolyticus.