A novel mechanism to ensure terminal initiation by hepatitis C virus NS5B polymerase

Virology. 2001 Jun 20;285(1):6-11. doi: 10.1006/viro.2001.0948.

Abstract

Hepatitis C virus (HCV) nonstructural protein 5B (NS5B) RNA-dependent RNA polymerase (RdRp) has acquired a unique beta-hairpin in the thumb subdomain which protrudes toward the active site. We report here that this beta-hairpin plays an important role in positioning the 3' terminus of the viral RNA genome for correct initiation of replication. The presence of this beta-hairpin interferes with polymerase binding to preannealed double-stranded RNA (dsRNA) molecules and allows only the single-stranded 3' terminus of an RNA template to bind productively to the active site. We propose that this beta-hairpin may serve as a "gate" which prevents the 3' terminus of the template RNA from slipping through the active site and ensures initiation of replication from the terminus of the genome. This hypothesis is supported by the ability of a beta-hairpin deletion mutant that utilizes dsRNA substrates and initiates RNA synthesis internally. The proposed terminal initiation mechanism may represent a novel replication strategy adopted by HCV and related viruses.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3' Untranslated Regions
  • Binding Sites
  • Hepacivirus / enzymology*
  • Hepatitis C / virology*
  • Point Mutation
  • RNA Replicase / chemistry
  • RNA Replicase / genetics*
  • RNA, Viral / biosynthesis*
  • Templates, Genetic
  • Transcription, Genetic
  • Viral Nonstructural Proteins / chemistry
  • Viral Nonstructural Proteins / genetics*
  • Virus Replication

Substances

  • 3' Untranslated Regions
  • NS-5 protein, hepatitis C virus
  • RNA, Viral
  • Viral Nonstructural Proteins
  • RNA Replicase