Identification of V. cholerae Type Three Secretion System Effector Proteins

AM-19226 is a pathogenic O39 serogroup Vibrio cholerae strain that lacks the typical virulence factors for colonization (TCP) and toxin production (CT), and instead encodes a Type III Secretion System (T3SS). The mechanism of pathogenesis is unknown, and few effector proteins have been identified. We therefore undertook a survey of the open reading frames within the ∼49.7 kb T3SS genomic island to identify potential effector proteins. We identified 15 ORFs for their ability to inhibit growth when expressed in yeast, and then used a β-lactamase (TEM1)-fusion reporter system to demonstrate that 11 proteins were bona fide effectors translocated into HeLa cells in vitro in a T3SS dependent manner. One effector, which we named VopX (A33_1663), is conserved only in V. cholerae and V. parahaemolyticus T3SS-positive strains and has not been previously studied. A vopX deletion reduces the ability of strain AM-19226 to colonize in vivo , and bile induced expression of a vopX-lacZ transcriptional fusion in vitro is regulated by the T3SS encoded transcriptional regulators VttR A and VttR B . An RLM1 yeast deletion strain rescued the growth inhibition induced by VopX expression, suggesting that VopX interacts with components of the cell wall integrity MAPK pathway. The collective results show that the V. cholerae T3SS encodes multiple effector proteins, one of which likely has novel activities that contribute to disease via interference with eukaryotic signaling pathways.