chemical genetics reveals bacterial and host cell functions critical for type iv effector translocation by legionella pneumophila化学遗传学显示细菌和宿主细胞功能的关键iv型效应由嗜肺性军团菌易位.pdfVIP

Chemical Genetics Reveals Bacterial and Host Cell Functions Critical for Type IV Effector Translocation by Legionella pneumophila 1 ¨ 1 1 2 2 Xavier Charpentier , Joelle E. Gabay , Moraima Reyes , Jing W. Zhu , Arthur Weiss , Howard A. Shuman1* 1 Department of Microbiology, Columbia University Medical Center, New York, New York, United States of America, 2 Departments of Medicine and of Microbiology and Immunology, Howard Hughes Medical Institute, Rosalind Russell Medical Research Center for Arthritis, University of California, San Francisco, California, United States of America Abstract Delivery of effector proteins is a process widely used by bacterial pathogens to subvert host cell functions and cause disease. Effector delivery is achieved by elaborate injection devices and can often be triggered by environmental stimuli. However, effector export by the L. pneumophila Icm/Dot Type IVB secretion system cannot be detected until the bacterium encounters a target host cell. We used chemical genetics, a perturbation strategy that utilizes small molecule inhibitors, to determine the mechanisms critical for L. pneumophila Icm/Dot activity. From a collection of more than 2,500 annotated molecules we identified specific inhibitors of effector translocation. We found that L. pneumophila effector translocation in macrophages requires host cell factors known to be involved in phagocytosis such as phosphoinositide 3-kinases, actin and tubulin. Moreover, we found that L. pneumophila phagocytosis and effector translocation also specifically require the receptor protein tyrosine phosphate phosphatases CD45 and CD148. We further show that phagocytosis is required to trigger effector delivery unless intimate contac