Visualization of the structures of the hepatitis C virus replication complex |
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Authors: | Chan Shih-Ching Lo Shih-Yen Liou Je-Wen Lin Min-Ching Syu Ciao-Ling Lai Meng-Jiun Chen Yi-Cheng Li Hui-Chun |
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Affiliation: | Graduate Institute of Molecular and Cellular Biology, Tzu Chi University, Hualien, Taiwan. |
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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. |
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