rna interference screen identifies abl kinase and pdgfr signaling in chlamydia trachomatis entryrna干扰屏幕识别abl激酶和pdgfr信号在沙眼衣原体条目.pdfVIP

RNA Interference Screen Identifies Abl Kinase and PDGFR Signaling in Chlamydia trachomatis Entry 1 1 1 2 1,3 Cherilyn A. Elwell , Alhaji Ceesay , Jung Hwa Kim , Daniel Kalman , Joanne N. Engel * 1 Department of Medicine, University of California San Francisco, San Francisco, California, United States of America, 2 Department of Pathology and Laboratory of Medicine, Emory University, Atlanta, Georgia, United States of America, 3 Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California, United States of America Abstract To elucidate the mechanisms involved in early events in Chlamydia trachomatis infection, we conducted a large scale unbiased RNA interference screen in Drosophila melanogaster S2 cells. This allowed identification of candidate host factors in a simple non-redundant, genetically tractable system. From a library of 7,216 double stranded RNAs (dsRNA), we identified ,226 host genes, including two tyrosine kinases, Abelson (Abl) kinase and PDGF- and VEGF-receptor related (Pvr), a homolog of the Platelet-derived growth factor receptor (PDGFR). We further examined the role of these two kinases in C. trachomatis binding and internalization into mammalian cells. Both kinases are phosphorylated upon infection and recruited to the site of bacterial attachment, but their roles in the infectious process are distinct. We provide evidence that PDGFRb may function as a receptor, as inhibition of PDGFRb by RNA interference or by PDGFRb neutralizing antibodies significantly reduces bacterial binding, whereas depletion of Abl kinase has no effect on binding