NS3 helicase domains involved in infectious intracellular hepatitis C virus particle assembly

Yinghong Ma, Jeremy Yates, Yuqiong Liang, Stanley M. Lemon, Min Kyung Yi

Research output: Contribution to journalArticle

157 Citations (Scopus)

Abstract

A mutation within subdomain 1 of the hepatitis C virus (HCV) NS3 helicase (NS3-Q221L) (M. Yi, Y. Ma, J. Yates, and S. M. Lemon, J. Virol. 81:629-638, 2007) rescues a defect in production of infectious virus by an intergenotypic chimeric RNA (HJ3). Although NS3-Gln-221 is highly conserved across HCV genotypes, the Leu-221 substitution had no effect on RNA replication or NS3-associated enzymatic activities. However, while transfection of unmodified HJ3 RNA failed to produce either extracellular or intracellular infectious virus, transfection of HJ3 RNA containing the Q221L substitution (HJ3/QL) resulted in rapid accumulation of intracellular infectious particles with release into extracellular fluids. In the absence of the Q221L mutation, both NS5A and NS3 were recruited to core protein on the surface of lipid droplets, but there was no assembly of core into high-density, rapidly sedimenting particles. Further analysis demonstrated that a Q221N mutation minimally rescued virus production and led to a second-site I399V mutation in subdomain 2 of the helicase. Similarly, I399V alone allowed only low-level virus production and led to selection of an I286V mutation in subdomain 1 of the helicase which fully restored virus production, confirming the involvement of both major helicase subdomains in the assembly process. Thus, multiple mutations in the helicase rescue a defect in an early-intermediate step in virus assembly that follows the recruitment of NS5A to lipid droplets and precedes the formation of dense intracellular viral particles. These data reveal a previously unsuspected role for the NS3 helicase in early virion morphogenesis and provide a new perspective on HCV assembly.

Original languageEnglish (US)
Pages (from-to)7624-7639
Number of pages16
JournalJournal of Virology
Volume82
Issue number15
DOIs
StatePublished - Aug 2008

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Virus Assembly
Hepatitis C virus
Hepatitis A
virion
Hepacivirus
Virion
mutation
Mutation
Viruses
viruses
RNA
transfection
droplets
Transfection
extracellular fluids
Extracellular Fluid
lipids
Morphogenesis
morphogenesis
Membrane Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

NS3 helicase domains involved in infectious intracellular hepatitis C virus particle assembly. / Ma, Yinghong; Yates, Jeremy; Liang, Yuqiong; Lemon, Stanley M.; Yi, Min Kyung.

In: Journal of Virology, Vol. 82, No. 15, 08.2008, p. 7624-7639.

Research output: Contribution to journalArticle

Ma, Yinghong ; Yates, Jeremy ; Liang, Yuqiong ; Lemon, Stanley M. ; Yi, Min Kyung. / NS3 helicase domains involved in infectious intracellular hepatitis C virus particle assembly. In: Journal of Virology. 2008 ; Vol. 82, No. 15. pp. 7624-7639.
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