Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus

Qingxia Han, Chunlian Xu, Chunchen Wu, Wenbo Zhu, Rongge Yang, Xinwen Chen

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

In this study, an infectious HCV monocistronic reporter virus was constructed by inserting an EGFP gene into the C-terminus of NS5A in the JFH-1 genome. A robust adaptive mutant, which could produce infectious virions as robustly as the JFH-1 wild type in Huh7.5.1 cells, was subsequently isolated by monitoring EGFP fluorescence. Full genomic sequencing revealed five amino acid substitutions, three located in the helicase domain of NS3 and two positioned in the C-terminus of NS5A. Reverse genetics studies suggested that the NS3 and NS5A mutations acted synergistically to enhance virus production capability possibly by accelerating the virion assembly efficiency but did not affect the replication competence of the adaptive reporter virus. Further analysis revealed that the M260K and T462I substitutions in NS3 and NS5A, respectively, were the key mutations. These adaptive mutations were also effective in the context of the JFH-1 genome.

Original languageEnglish (US)
Pages (from-to)63-73
Number of pages11
JournalVirus Research
Volume145
Issue number1
DOIs
StatePublished - Oct 1 2009
Externally publishedYes

Fingerprint

Hepacivirus
Viruses
Virion
Mutation
Genome
Reverse Genetics
Proteins
Amino Acid Substitution
Mental Competency
Fluorescence
Genes

Keywords

  • Adaptive Mutation
  • Assembly
  • HCV
  • Hepatitis C virus
  • Reporter Virus

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

Cite this

Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus. / Han, Qingxia; Xu, Chunlian; Wu, Chunchen; Zhu, Wenbo; Yang, Rongge; Chen, Xinwen.

In: Virus Research, Vol. 145, No. 1, 01.10.2009, p. 63-73.

Research output: Contribution to journalArticle

Han, Qingxia ; Xu, Chunlian ; Wu, Chunchen ; Zhu, Wenbo ; Yang, Rongge ; Chen, Xinwen. / Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus. In: Virus Research. 2009 ; Vol. 145, No. 1. pp. 63-73.
@article{71c92ddebf9646678a2a0df6c8239c35,
title = "Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus",
abstract = "In this study, an infectious HCV monocistronic reporter virus was constructed by inserting an EGFP gene into the C-terminus of NS5A in the JFH-1 genome. A robust adaptive mutant, which could produce infectious virions as robustly as the JFH-1 wild type in Huh7.5.1 cells, was subsequently isolated by monitoring EGFP fluorescence. Full genomic sequencing revealed five amino acid substitutions, three located in the helicase domain of NS3 and two positioned in the C-terminus of NS5A. Reverse genetics studies suggested that the NS3 and NS5A mutations acted synergistically to enhance virus production capability possibly by accelerating the virion assembly efficiency but did not affect the replication competence of the adaptive reporter virus. Further analysis revealed that the M260K and T462I substitutions in NS3 and NS5A, respectively, were the key mutations. These adaptive mutations were also effective in the context of the JFH-1 genome.",
keywords = "Adaptive Mutation, Assembly, HCV, Hepatitis C virus, Reporter Virus",
author = "Qingxia Han and Chunlian Xu and Chunchen Wu and Wenbo Zhu and Rongge Yang and Xinwen Chen",
year = "2009",
month = "10",
day = "1",
doi = "10.1016/j.virusres.2009.06.005",
language = "English (US)",
volume = "145",
pages = "63--73",
journal = "Virus Research",
issn = "0168-1702",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus

AU - Han, Qingxia

AU - Xu, Chunlian

AU - Wu, Chunchen

AU - Zhu, Wenbo

AU - Yang, Rongge

AU - Chen, Xinwen

PY - 2009/10/1

Y1 - 2009/10/1

N2 - In this study, an infectious HCV monocistronic reporter virus was constructed by inserting an EGFP gene into the C-terminus of NS5A in the JFH-1 genome. A robust adaptive mutant, which could produce infectious virions as robustly as the JFH-1 wild type in Huh7.5.1 cells, was subsequently isolated by monitoring EGFP fluorescence. Full genomic sequencing revealed five amino acid substitutions, three located in the helicase domain of NS3 and two positioned in the C-terminus of NS5A. Reverse genetics studies suggested that the NS3 and NS5A mutations acted synergistically to enhance virus production capability possibly by accelerating the virion assembly efficiency but did not affect the replication competence of the adaptive reporter virus. Further analysis revealed that the M260K and T462I substitutions in NS3 and NS5A, respectively, were the key mutations. These adaptive mutations were also effective in the context of the JFH-1 genome.

AB - In this study, an infectious HCV monocistronic reporter virus was constructed by inserting an EGFP gene into the C-terminus of NS5A in the JFH-1 genome. A robust adaptive mutant, which could produce infectious virions as robustly as the JFH-1 wild type in Huh7.5.1 cells, was subsequently isolated by monitoring EGFP fluorescence. Full genomic sequencing revealed five amino acid substitutions, three located in the helicase domain of NS3 and two positioned in the C-terminus of NS5A. Reverse genetics studies suggested that the NS3 and NS5A mutations acted synergistically to enhance virus production capability possibly by accelerating the virion assembly efficiency but did not affect the replication competence of the adaptive reporter virus. Further analysis revealed that the M260K and T462I substitutions in NS3 and NS5A, respectively, were the key mutations. These adaptive mutations were also effective in the context of the JFH-1 genome.

KW - Adaptive Mutation

KW - Assembly

KW - HCV

KW - Hepatitis C virus

KW - Reporter Virus

UR - http://www.scopus.com/inward/record.url?scp=69449083296&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69449083296&partnerID=8YFLogxK

U2 - 10.1016/j.virusres.2009.06.005

DO - 10.1016/j.virusres.2009.06.005

M3 - Article

C2 - 19540283

AN - SCOPUS:69449083296

VL - 145

SP - 63

EP - 73

JO - Virus Research

JF - Virus Research

SN - 0168-1702

IS - 1

ER -