Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis

Hamed Gouklani, Rowena A Bull, Claudia Beyer, Fasseli J Coulibaly, Eric J Gowans, Heidi E Drummer, Hans J Netter, Peter A White, Gholamreza Haqshenas

Research output: Contribution to journalArticleResearchpeer-review

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.
Original languageEnglish
Pages (from-to)5080 - 5088
Number of pages9
JournalJournal of Virology
Volume86
Issue number9
DOIs
Publication statusPublished - 2012

Cite this

Gouklani, Hamed ; Bull, Rowena A ; Beyer, Claudia ; Coulibaly, Fasseli J ; Gowans, Eric J ; Drummer, Heidi E ; Netter, Hans J ; White, Peter A ; Haqshenas, Gholamreza. / Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis. In: Journal of Virology. 2012 ; Vol. 86, No. 9. pp. 5080 - 5088.
@article{d22f388e859743f1a45cdff318780eb8,
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.",
author = "Hamed Gouklani and Bull, {Rowena A} and Claudia Beyer and Coulibaly, {Fasseli J} and Gowans, {Eric J} and Drummer, {Heidi E} and Netter, {Hans J} and White, {Peter A} and Gholamreza Haqshenas",
year = "2012",
doi = "10.1128/JVI.07089-11",
language = "English",
volume = "86",
pages = "5080 -- 5088",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "9",

}

Gouklani, H, Bull, RA, Beyer, C, Coulibaly, FJ, Gowans, EJ, Drummer, HE, Netter, HJ, White, PA & Haqshenas, G 2012, 'Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis' Journal of Virology, vol. 86, no. 9, pp. 5080 - 5088. https://doi.org/10.1128/JVI.07089-11

Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis. / Gouklani, Hamed; Bull, Rowena A; Beyer, Claudia; Coulibaly, Fasseli J; Gowans, Eric J; Drummer, Heidi E; Netter, Hans J; White, Peter A; Haqshenas, Gholamreza.

In: Journal of Virology, Vol. 86, No. 9, 2012, p. 5080 - 5088.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Hepatitis C virus nonstructural protein 5B is involved in virus morphogenesis

AU - Gouklani, Hamed

AU - Bull, Rowena A

AU - Beyer, Claudia

AU - Coulibaly, Fasseli J

AU - Gowans, Eric J

AU - Drummer, Heidi E

AU - Netter, Hans J

AU - White, Peter A

AU - Haqshenas, Gholamreza

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

UR - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3347352/pdf/zjv5080.pdf

U2 - 10.1128/JVI.07089-11

DO - 10.1128/JVI.07089-11

M3 - Article

VL - 86

SP - 5080

EP - 5088

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 9

ER -