title: Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis.
abstract: The p7 protein of hepatitis C virus (HCV) is a viroporin that is dispensable for viral genome replication but plays a critical role in virus morphogenesis. In this study, we generated a JFH1-based intergenotypic chimeric genome that encoded a heterologous genotype 1b (GT1b) p7. The parental intergenotypic chimeric genome was nonviable in human hepatoma cells, and infectious chimeric virions were produced only when cells transfected with the chimeric genomes were passaged several times. Sequence analysis of the entire polyprotein-coding region of the recovered chimeric virus revealed one predominant amino acid substitution in nonstructural protein 2 (NS2), T23N, and one in NS5B, K151R. Forward genetic analysis demonstrated that each of these mutations per se restored the infectivity of the parental chimeric genome, suggesting that interactions between p7, NS2, and NS5B were required for virion assembly/maturation. p7 and NS5B colocalized in cellular compartments, and the NS5B mutation did not affect the colocalization pattern. The NS5B K151R mutation neither increased viral RNA replication in human hepatoma cells nor altered the polymerase activity of NS5B in an in vitro assay. In conclusion, this study suggests that HCV NS5B is involved in virus morphogenesis.
authors.affiliation: The Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria, Australia.
authors.forename: Hamed
authors.initials: H
authors.lastname: Gouklani
authors.forename: Rowena A
authors.initials: RA
authors.lastname: Bull
authors.forename: Claudia
authors.initials: C
authors.lastname: Beyer
authors.forename: Fasséli
authors.initials: F
authors.lastname: Coulibaly
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Gowans
authors.forename: Heidi E
authors.initials: HE
authors.lastname: Drummer
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Netter
authors.forename: Peter A
authors.initials: PA
authors.lastname: White
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Haqshenas
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002384:Catalysis; D002460:Cell Line; D016679:Genome, Viral; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D011487:Protein Conformation; D021381:Protein Transport; D012367:RNA, Viral / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D019065:Virus Assembly / Q000235:genetics; D014779:Virus Replication / Q000235:genetics; D000086082:Viroporin Proteins; D012324:RNA-Dependent RNA Polymerase
chemical_list: D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C097010:NS2 protein, Hepatitis C virus; C088369:p7 protein, Hepatitis C virus; D000086082:Viroporin Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase