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DAF and other GPI-anchored proteins can be found in cholesterol-rich ordered domains within the plasma membrane that are known as \u201clipid rafts.\u201d We have shown, by using drugs to specifically inhibit various endocytosis routes, that infection by a DAF-using strain of echovirus 11 (EV11) is dependent upon cholesterol and an intact cytoskeleton, whereas a non-DAF-using mutant derived from it was unaffected by these drugs. Using RNA transfection and virus-binding assays, we have shown that this requirement for cholesterol, the actin cytoskeleton, and the microtubule network occurs postbinding of the virus but prior to uncoating of the RNA, indicating a role during virus entry. Confocal microscopy of virus infection supported the role of cholesterol and the cytoskeleton during entry. In addition, [<jats:sup>35<\/jats:sup>S]methionine-labeled DAF-using EV11, but not the non-DAF-using EV11, could be copurified with lipid raft components during infection after Triton X-100 extraction. These data indicate that DAF usage by EV11 enables the virus to associate with lipid rafts and enter cells through this novel route.<\/jats:p>","DOI":"10.1128\/jvi.76.18.9307-9322.2002","type":"journal-article","created":{"date-parts":[[2002,10,1]],"date-time":"2002-10-01T19:36:19Z","timestamp":1033500979000},"page":"9307-9322","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":106,"title":["A Novel Cell Entry Pathway for a DAF-Using Human Enterovirus Is Dependent on Lipid Rafts"],"prefix":"10.1128","volume":"76","author":[{"given":"Amanda D.","family":"Stuart","sequence":"first","affiliation":[{"name":"Division of Virology, Department of Pathology, University of Cambridge, CB2 1QP Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah E.","family":"Eustace","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Pathology, University of Cambridge, CB2 1QP Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas A.","family":"McKee","sequence":"additional","affiliation":[{"name":"Institute Universitaire de Pathologie, CH-1011 Lausanne, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T. 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