title: The second extracellular loop dictates Occludin-mediated HCV entry.
abstract: Recent findings have implicated tight junction (TJ) protein Occludin (OCLN) as an essential factor for hepatitis C virus (HCV) to enter human hepatocytes. To gain insights into OCLN-mediated HCV entry, we created a panel of OCLN deletion mutants and found that without impairing OCLN's cell surface localization, removal of the extracellular loop 2 (EL2) from OCLN abolished both its ability to mediate HIV-HCV pseudotypes' (HCVpp) entry as well as its ability to coprecipitate HCV glycoprotein E2. Recombinant OCLN EL2, however, failed to robustly bind soluble E2 (sE2) in pull-down assays. Subsequent studies revealed that OCLN formed complex with Dynamin II, an important GTPase for endocytosis, in an EL2-dependent fashion. HCVpp, as well as cell culture grown HCV (HCVcc), was sensitive to Dynamin knockdown or inhibition. We conclude that OCLN EL2 dictates the Dynamin-dependent HCV entry. Furthermore, OCLN could function to bridge virions to Dynamin-dependent endocytic machineries.
authors.affiliation: Department of Infectious Diseases and Microbiology, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Shufeng
authors.initials: S
authors.lastname: Liu
authors.forename: Wayne
authors.initials: W
authors.lastname: Kuo
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.forename: Weiqun
authors.initials: W
authors.lastname: Liu
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Gibson
authors.forename: Kenneth
authors.initials: K
authors.lastname: Dorko
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Strom
authors.forename: Tianyi
authors.initials: T
authors.lastname: Wang
mesh_terms: D002460:Cell Line; D034285:Dynamin II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006678:HIV / Q000235:genetics; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D050505:Mutant Proteins / Q000235:genetics / Q000502:physiology; D062793:Occludin; D011485:Protein Binding; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology*; D017384:Sequence Deletion; D053586:Virus Internalization
chemical_list: D008565:Membrane Proteins; D050505:Mutant Proteins; C568985:OCLN protein, human; D062793:Occludin; D011991:Receptors, Virus; D034285:Dynamin II
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