Ross-Thriepland, D, Amako, Y and Harris, M (2013) The C terminus of NS5A domain II is a key determinant of hepatitis C virus genome replication, but is not required for virion assembly and release. Journal of General Virology, 94 (Pt 5). 1009 - 1018. ISSN 0022-1317
Abstract
The NS5A protein of hepatitis C virus (HCV) plays roles in both virus genome replication and the assembly of infectious virus particles. NS5A comprises three domains, separated by low-complexity sequences. Whilst the function of domain I appears to be predominantly involved with genome replication, the roles of domains II and III are less well defined. It has been reported previously that a deletion spanning the majority of domain II but retaining the C-terminal 35 residues had no effect on virus production; however, deletion of the entire domain II eliminated genome replication, pointing to a key role for the C terminus of this domain. Recent work has also highlighted this region as the potential binding site of the host factor cyclophilin A (CypA). To define this requirement for replication in more detail, and to investigate the involvement of CypA, we conducted a mutagenic study of the C-terminal 30 residues of domain II within the context of both the infectious JFH-1 virus and a JFH-1-derived subgenomic replicon. We showed that 12 of these residues were absolutely required for virus genome replication, whilst mutations of the remainder either had no phenotype or exhibited a partial reduction in genome replication. There was an absolute correlation between the datasets for virus and subgenomic replicon, indicating that this region is involved solely in the process of genome replication. Comparison of our data with a previously published analysis of the same region in genotype 1b revealed some important differences between the two genotypes of HCV.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2013 SGM. This is an open access article published by the Microbiology Society under the Creative Commons Attribution License. |
Keywords: | Amino Acid Motifs; Amino Acid Sequence; Amino Acid Substitution; Cell Line, Tumor; Consensus Sequence; Cyclophilin A; Genotype; Hepacivirus; Humans; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Phenotype; Protein Binding; Protein Structure, Tertiary; RNA, Viral; Sequence Analysis, DNA; Viral Nonstructural Proteins; Virus Assembly; Virus Replication |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 11 Nov 2015 16:22 |
Last Modified: | 11 Nov 2015 16:22 |
Published Version: | http://dx.doi.org/10.1099/vir.0.050633-0 |
Status: | Published |
Publisher: | Society for General Microbiology |
Identification Number: | 10.1099/vir.0.050633-0 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:85129 |