title: Visualization of the structures of the hepatitis C virus replication complex.
abstract: Hepatitis C viral RNA synthesis has been demonstrated to occur on a lipid raft membrane structure. Lipid raft membrane fraction purified by membrane flotation analysis was observed using transmission electron microscopy and atomic force microscopy. Particles around 0.7 um in size were found in lipid raft membrane fraction purified from hepatitis C virus (HCV) replicon but not their parental HuH7 cells. HCV NS5A protein was associated with these specialized particles. After several cycles of freezing-thawing, these particles would fuse into larger sizes up to 10 um. Knockdown of seven proteins associated with lipid raft (VAPA, COPG, RAB18, COMT, CDC42, DPP4, and KDELR2) of HCV replicon cells reduced the observed number of these particles and suppressed the HCV replication. Results in this study indicated that HCV replication complexes with associated lipid raft membrane form distinct particle structures of around 0.7 um as observed from transmission electron microscopy and atomic force microscopy.
authors.affiliation: Graduate Institute of Molecular and Cellular Biology, Tzu Chi University, Hualien, Taiwan.
authors.forename: Shih-Ching
authors.initials: SC
authors.lastname: Chan
authors.forename: Shih-Yen
authors.initials: SY
authors.lastname: Lo
authors.forename: Je-Wen
authors.initials: JW
authors.lastname: Liou
authors.forename: Min-Ching
authors.initials: MC
authors.lastname: Lin
authors.forename: Ciao-Ling
authors.initials: CL
authors.lastname: Syu
authors.forename: Meng-Jiun
authors.initials: MJ
authors.lastname: Lai
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Chen
authors.forename: Hui-Chun
authors.initials: HC
authors.lastname: Li
mesh_terms: D002460:Cell Line; D003902:Detergents / Q000737:chemistry; D055785:Gene Knockdown Techniques; D016174:Hepacivirus / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism / Q000821:virology*; D046529:Microscopy, Electron, Transmission; D034741:RNA, Small Interfering / Q000235:genetics; D012093:Replicon; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: D003902:Detergents; D034741:RNA, Small Interfering; D014764:Viral Proteins