A genomic analysis of this organism revealed two sets of type III secretion systems, T3SS1 and T3SS2 (Makino check details et al., 2003), and functional assays were carried out to examine the contribution of each T3SS to the pathogenicity of V. parahaemolyticus (Park et al., 2004; Ono et al., 2006; Hiyoshi et al., 2010; Pineyro et al., 2010). The results indicated that the enterotoxicity of this bacterium in humans was dependent on T3SS2. The genes encoding for T3SS2 are located within the V. parahaemolyticus pathogenicity island (Vp-PAI) (Sugiyama et al., 2008) that

causes fluid accumulation in a rabbit ileal loop model (Park et al., 2004; Hiyoshi et al., 2010), and it has been confirmed that T3SS2 causes diarrhea in a piglet model (Pineyro et al., ICG-001 molecular weight 2010). Many Gram-negative bacteria utilize the T3SS to efficiently manipulate their hosts by injecting virulence factors, so-called effectors, into host cells (Coburn et al., 2007; Galan, 2009). Protein secretion by T3SS is co-operatively regulated by the control of transcription of T3SS effectors/components, and at the post-transcriptional level (Francis et al., 2002; Yahr & Wolfgang, 2006). Previous studies have shown that the T3SS effector/chaperone complex is indispensable for the efficient delivery of effectors into host cells (Galan & Wolf-Watz, 2006), as hypothesized in the model of the protein secretion mechanism

(Arnold et al., 2009). The established model is based on a single T3SS apparatus present

Palmatine in one bacterium, and questions have arisen as to how the destination of effectors is determined in a bacterium equipped with multiple T3SSs. There are several bacteria with multiple T3SSs, including Salmonella (Knodler et al., 2002), enterohemorrhagic Escherichia coli (Hartleib et al., 2003), Burkholderia pseudomallei (Attree & Attree, 2001), and V. parahaemolyticus (Makino et al., 2003). Of these, V. parahaemolyticus is the best model for exploring the specificity of protein secretion mechanisms in the presence of multiple T3SSs because V. parahaemolyticus can specifically secrete multiple effectors via two individual T3SSs under the same culture conditions (Akeda et al., 2009). Based on the current model of protein secretion through the T3SS, T3SS-specific chaperones or the amino-terminal secretion signal sequence of secreted effectors could be the determinant of the specificity of effector secretion via individual apparatuses (Arnold et al., 2009). The specificity of effector secretion through Salmonella pathogenicity island-1 (SPI-1) or the flagellar system is dependent on the T3SS chaperones of the secreted effectors (Lee & Galan, 2004). However, the requirements for specificity in nonflagellar-type T3SSs for the secretion of T3SS effectors in the same bacterial cell have not been investigated. In V. parahaemolyticus, there are a number of T3SS1- and 2-specific effectors. The T3SS2-specific effectors include VopP (Park et al.