The marine bacterium causes food-borne diseases, which may lead to life-threatening septicemia in some individuals. Therefore, identifying virulence factors in is of high priority. We performed a transcriptome analysis on after infection of human intestinal HT29-methotrexate cells and found induction of, encoding a putative phospholipase, PlpA. Bioinformatics, biochemical, and genetic analyses demonstrated thatPlpA is a phospholipase A secreted in a type II secretion system-dependent manner. Compared with the wild type, the mutant exhibited reduced mortality, systemic infection, and inflammation in mice as well as low cytotoxicity toward the human epithelial INT-407 cells. Moreover, mutation attenuated the release of actin and cytosolic cyclophilin A from INT-407 cells, indicating that PlpA is a virulence factor essential for causing lysis and necrotic death of the epithelial cells. transcription was growth phase-dependent, reaching maximum levels during the early stationary phase. Also, transcription factor HlyU and cAMP receptor protein (CRP) mediate additive activation and host-dependent induction of Molecular biological analyses revealed that expression is controlled via the promoter, P , and that HlyU and CRP directly bind to P upstream sequences. Taken together, this study demonstrated that PlpA is a type II secretion system-dependent secretory phospholipase A regulated by HlyU and CRP and is essential for the pathogenicity of .