Association of hepatitis C virus replication complexes with microtubules and actin filaments is dependent on the interaction of NS3 and NS5A.
The hepatitis C virus (HCV) RNA replication complex (RC), which is composed of viral nonstructural (NS) proteins and host cellular proteins, replicates the viral RNA genome in association with intracellular membranes. Two viral NS proteins, NS3 and NS5A, are essential elements of the RC. Here, by using immunoprecipitation and fluorescence resonance energy transfer assays, we demonstrated that NS3 and NS5A interact with tubulin and actin. Furthermore, immunofluorescence microscopy and electron microscopy revealed that HCV RCs were aligned along microtubules and actin filaments in both HCV replicon cells and HCV-infected cells. In addition, the movement of RCs was inhibited when microtubules or actin filaments were depolymerized by colchicine and cytochalasin B, respectively. Based on our observations, we propose that microtubules and actin filaments provide the tracks for the movement of HCV RCs to other regions in the cell, and the molecular interactions between RCs and microtubules, or RCs and actin filaments, are mediated by NS3 and NS5A.
Lai
Chao-Kuen
CK
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
Jeng
King-Song
KS
Machida
Keigo
K
Lai
Michael M C
MM
D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000911:Antibodies, Monoclonal / Q000276:immunology; D002232:Carbocyanines / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D019085:Fluorescent Antibody Technique, Direct; D005456:Fluorescent Dyes / Q000378:metabolism; D016174:Hepacivirus / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000378:metabolism; D007668:Kidney / Q000166:cytology; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D020365:RNA Helicases / Q000378:metabolism / Q000648:ultrastructure; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012093:Replicon; D012697:Serine Endopeptidases / Q000378:metabolism / Q000648:ultrastructure; D014404:Tubulin / Q000378:metabolism; D050257:Tubulin Modulators / Q000494:pharmacology; D017361:Viral Nonstructural Proteins / Q000378:metabolism* / Q000648:ultrastructure; D014779:Virus Replication; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000911:Antibodies, Monoclonal; D002232:Carbocyanines; D003078:Colchicine; D003571:Cytochalasin B; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D007211:Indoles; D020365:RNA Helicases; D012367:RNA, Viral; D012697:Serine Endopeptidases; D014404:Tubulin; D050257:Tubulin Modulators; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C096041:NS5 protein, flavivirus; C089187:cyanine dye 3; C007293:DAPI