Stimulation of hepatitis C virus (HCV) nonstructural protein 3 (NS3) helicase activity by the NS3 protease domain and by HCV RNA-dependent RNA polymerase

Chen Zhang, Zhaohui Cai, Young Chan Kim, Ranjith Kumar, Fenghua Yuan, Pei-Yong Shi, Cheng Kao, Guangxiang Luo

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Hepatitis C virus (HCV) nonstructural protein 3 (NS3) possesses multiple enzyme activities. The N-terminal one-third of NS3 primarily functions as a serine protease, while the remaining two-thirds of NS3 serve as a helicase and nucleoside triphosphatase. Whether the multiple enzyme activities of NS3 are functionally interdependent and/or modulated by other viral NS proteins remains unclear. We performed biochemical studies to examine the functional interdependence of the NS3 protease and helicase domains and the modulation of NS3 helicase by NS5B, an RNA-dependent RNA polymerase (RdRp). We found that the NS3 protease domain of the full-length NS3 (NS3FL) enhances the NS3 helicase activity. Additionally, HCV RdRp stimulates the NS3FL helicase activity by more than sevenfold. However, the helicase activity of the NS3 helicase domain was unaffected by HCV RdRp. Glutathione S-transferase pull-down as well as fluorescence anisotropy results revealed that the NS3 protease domain is required for specific NS3 and NS5B interaction. These findings suggest that HCV RdRp regulates the functions of NS3 during HCV replication. In contrast, NS3FL does not increase NS5B RdRp activity in vitro, which is contrary to a previously published report that the HCV NS3 enhances NS5B RdRp activity.

Original languageEnglish (US)
Pages (from-to)8687-8697
Number of pages11
JournalJournal of Virology
Volume79
Issue number14
DOIs
StatePublished - Jul 2005
Externally publishedYes

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RNA-directed RNA polymerase
RNA Replicase
Hepatitis C virus
Hepacivirus
Peptide Hydrolases
proteinases
Proteins
proteins
Viral Nonstructural Proteins
Nucleoside-Triphosphatase
enzyme activity
Fluorescence Polarization

ASJC Scopus subject areas

  • Immunology

Cite this

Stimulation of hepatitis C virus (HCV) nonstructural protein 3 (NS3) helicase activity by the NS3 protease domain and by HCV RNA-dependent RNA polymerase. / Zhang, Chen; Cai, Zhaohui; Kim, Young Chan; Kumar, Ranjith; Yuan, Fenghua; Shi, Pei-Yong; Kao, Cheng; Luo, Guangxiang.

In: Journal of Virology, Vol. 79, No. 14, 07.2005, p. 8687-8697.

Research output: Contribution to journalArticle

Zhang, Chen ; Cai, Zhaohui ; Kim, Young Chan ; Kumar, Ranjith ; Yuan, Fenghua ; Shi, Pei-Yong ; Kao, Cheng ; Luo, Guangxiang. / Stimulation of hepatitis C virus (HCV) nonstructural protein 3 (NS3) helicase activity by the NS3 protease domain and by HCV RNA-dependent RNA polymerase. In: Journal of Virology. 2005 ; Vol. 79, No. 14. pp. 8687-8697.
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