a cytotoxic type iii secretion effector of vibrio parahaemolyticus targets vacuolar h+-atpase subunit c and ruptures host cell lysosomes的细胞毒性iii型分泌效应弧菌parahaemolyticus目标空泡的h + atp酶亚基c和破裂宿主细胞溶酶体.pdfVIP

A Cytotoxic Type III Secretion Effector of Vibrio parahaemolyticus + Targets Vacuolar H -ATPase Subunit c and Ruptures Host Cell Lysosomes 1 1¤ 2 2 1 Shigeaki Matsuda , Natsumi Okada , Toshio Kodama , Takeshi Honda , Tetsuya Iida * 1 Laboratory of Genomic Research on Pathogenic Bacteria, International Research Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan, 2 Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan Abstract Vibrio parahaemolyticus is one of the human pathogenic vibrios. During the infection of mammalian cells, this pathogen exhibits cytotoxicity that is dependent on its type III secretion system (T3SS1). VepA, an effector protein secreted via the T3SS1, plays a major role in the T3SS1-dependent cytotoxicity of V. parahaemolyticus. However, the mechanism by which VepA is involved in T3SS1-dependent cytotoxicity is unknown. Here, we found that protein transfection of VepA into HeLa cells resulted in cell death, indicating that VepA alone is cytotoxic. The ectopic expression of VepA in yeast Saccharomyces cerevisiae interferes with yeast growth, indicating that VepA is also toxic in yeast. A yeast genome-wide screen identified the yeast gene VMA3 as essential for the growth inhibition of yeast by VepA. Although VMA3 encodes subunit c of the vacuolar H+-ATPase (V-ATPase), the toxicity of VepA was independent of the function of V-ATPases. In HeLa cells, knockdown of V- ATPase subunit c decreased VepA-mediated cytotoxicity. We al