title: Conserved glycine 33 residue in flexible domain I of hepatitis C virus core protein is critical for virus infectivity.
abstract: Hepatitis C virus core protein forms the viral nucleocapsid and plays a critical role in the formation of infectious particles. In this study, we demonstrate that the highly conserved residue G33, located within domain 1 of the core protein, is important for the production of cell culture-infectious virus (HCVcc). Alanine substitution at this position in the JFH1 genome did not alter viral RNA replication but reduced infectivity by ∼2 logs. Virus production by this core mutant could be rescued by compensatory mutations located immediately upstream and downstream of the original G33A mutation. The examination of the helix-loop-helix motif observed in the core protein structure (residues 15 to 41; Protein Data Bank entry 1CWX) indicated that the residues G33 and F24 are in close contact with each other, and that the G33A mutation induces a steric clash with F24. Molecular simulations revealed that the compensatory mutations increase the helix-loop-helix flexibility, allowing rescue of the core active conformation required for efficient virus production. Taken together, these data highlight the plasticity of core domain 1 conformation and illustrate the relationship between its structural tolerance to mutations and virus infectivity.
authors.affiliation: MRC--University of Glasgow Centre for Virus Research, Glasgow, United Kingdom.
authors.forename: Allan G N
authors.initials: AG
authors.lastname: Angus
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.forename: Séamus J
authors.initials: SJ
authors.lastname: Stack
authors.forename: David
authors.initials: D
authors.lastname: Dalrymple
authors.forename: Derek
authors.initials: D
authors.lastname: Gatherer
authors.forename: François
authors.initials: F
authors.lastname: Penin
authors.forename: Arvind H
authors.initials: AH
authors.lastname: Patel
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D017124:Conserved Sequence; D005998:Glycine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016174:Hepacivirus / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D014758:Viral Core Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D005998:Glycine
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