Structure of the zinc-binding domain of an essential component of the hepatitis C virus replicase

Nature. 2005 May 19;435(7040):374-9. doi: 10.1038/nature03580.

Abstract

Hepatitis C virus (HCV) is a human pathogen affecting nearly 3% of the world's population. Chronic infections can lead to cirrhosis and liver cancer. The RNA replication machine of HCV is a multi-subunit membrane-associated complex. The non-structural protein NS5A is an active component of HCV replicase, as well as a pivotal regulator of replication and a modulator of cellular processes ranging from innate immunity to dysregulated cell growth. NS5A is a large phosphoprotein (56-58 kDa) with an amphipathic alpha-helix at its amino terminus that promotes membrane association. After this helix region, NS5A is organized into three domains. The N-terminal domain (domain I) coordinates a single zinc atom per protein molecule. Mutations disrupting either the membrane anchor or zinc binding of NS5A are lethal for RNA replication. However, probing the role of NS5A in replication has been hampered by a lack of structural information about this multifunctional protein. Here we report the structure of NS5A domain I at 2.5-A resolution, which contains a novel fold, a new zinc-coordination motif and a disulphide bond. We use molecular surface analysis to suggest the location of protein-, RNA- and membrane-interaction sites.

Publication types

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

MeSH terms

  • Amino Acid Motifs
  • Binding Sites
  • Crystallography, X-Ray
  • Dimerization
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Hepacivirus / enzymology*
  • Hepacivirus / genetics
  • Models, Molecular
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary
  • RNA Replicase / chemistry*
  • RNA Replicase / metabolism
  • Viral Nonstructural Proteins / chemistry*
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism*
  • Zinc / metabolism*

Substances

  • Disulfides
  • NS-5 protein, hepatitis C virus
  • Viral Nonstructural Proteins
  • RNA Replicase
  • Zinc

Associated data

  • PDB/1ZH1