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Here we present a systematic study of the internalization mechanism of JEV in fibroblasts and neuroblastoma cells. To verify the roles of distinct pathways of cell entry, we used fluorescently labeled virus particles, a combination of pharmacological inhibitors, RNA interference (RNAi), and dominant-negative (DN) mutants of regulatory proteins involved in endocytosis. Our study demonstrates that JEV infects fibroblasts in a clathrin-dependent manner, but it deploys a clathrin-independent mechanism to infect neuronal cells. The clathrin-independent pathway requires dynamin and plasma membrane cholesterol. Virus binding to neuronal cells leads to rapid actin rearrangements and an intact and dynamic actin cytoskeleton, and the small GTPase RhoA plays an important role in viral entry. Immunofluorescence analysis of viral colocalization with endocytic markers showed that JEV traffics through Rab5-positive early endosomes and that release of the viral nucleocapsid occurs at the level of the early and not the late endosomes.<\/jats:p>","DOI":"10.1128\/jvi.01399-12","type":"journal-article","created":{"date-parts":[[2012,10,11]],"date-time":"2012-10-11T09:01:29Z","timestamp":1349946089000},"page":"148-162","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":139,"title":["Japanese Encephalitis Virus Infects Neuronal Cells through a Clathrin-Independent Endocytic Mechanism"],"prefix":"10.1128","volume":"87","author":[{"given":"Manjula","family":"Kalia","sequence":"first","affiliation":[{"name":"Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renu","family":"Khasa","sequence":"additional","affiliation":[{"name":"Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manish","family":"Sharma","sequence":"additional","affiliation":[{"name":"Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minu","family":"Nain","sequence":"additional","affiliation":[{"name":"Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sudhanshu","family":"Vrati","sequence":"additional","affiliation":[{"name":"Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, Gurgaon, Haryana, India"},{"name":"National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"1101","volume-title":"Fields virology","author":"Lindenbach BD","year":"2007","unstructured":"LindenbachBD Thiel Heinz-Jurgen RiceCM. 2007. 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