rnai screen reveals an abl kinase-dependent host cell pathway involved in pseudomonas aeruginosa internalizationrnai屏幕显示一个abl kinase-dependent宿主细胞通路参与铜绿假单胞菌内化.pdfVIP

RNAi Screen Reveals an Abl Kinase-Dependent Host Cell Pathway Involved in Pseudomonas aeruginosa Internalization 1,2,3 4 4 1,2,3 Julia F. Pielage , Kimberly R. Powell , Daniel Kalman , Joanne N. Engel * 1 Program in Microbial Pathogenesis and Host Defense, University of California San Francisco, San Francisco, California, United States of America, 2 Department of Medicine, University of California San Francisco, San Francisco, California, United States of America, 3 Department of Microbiology Immunology, University of California San Francisco, San Francisco, California, United States of America, 4 Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, United States of America Abstract Internalization of the pathogenic bacterium Pseudomonas aeruginosa by non-phagocytic cells is promoted by rearrangements of the actin cytoskeleton, but the host pathways usurped by this bacterium are not clearly understood. We used RNAi-mediated gene inactivation of ,80 genes known to regulate the actin cytoskeleton in Drosophila S2 cells to identify host molecules essential for entry of P. aeruginosa. This work revealed Abl tyrosine kinase, the adaptor protein Crk, the small GTPases Rac1 and Cdc42, and p21-activated kinase as components of a host signaling pathway that leads to internalization of P. aeruginosa. Using a variety of complementary approaches, we validated the role of this pathway in mammalian cells. Remarkably, ExoS and ExoT, type III secreted toxins of P. aeruginosa, target this pathway by interfering with GTPase function and, in the case of ExoT, by abrogating P. aeruginosa–induced Abl-dependent Crk phosphorylation.