Processing of Norwalk virus nonstructural proteins by a 3C-like cysteine proteinase

Virology. 2003 Apr 10;308(2):216-24. doi: 10.1016/s0042-6822(03)00004-7.

Abstract

Expression of Norwalk virus nonstructural polyprotein precursor in vitro resulted in rapid cotranslational cleavage at specific sites. The cleavage products were similar to those previously identified for Southampton virus, a highly related virus. We inactivated the virally encoded proteinase responsible for cleavage of the nonstructural polyprotein by mutation of the putative catalytic cysteine residue, which resulted in production of full-length polyprotein precursor. NV proteinase was expressed in Escherichia coli as a glutathione S-transferase fusion and purified by GST-affinity chromatography. Activity of the purified proteinase was demonstrated by incubation with the full-length precursor protein. trans cleavage of the nonstructural protein precursor resulted in cleavage products similar to those observed during cotranslational cleavage, however, at lesser efficiency. NV proteinase displayed sensitivities to cysteine and serine protease inhibitors similar to poliovirus 3C proteinase, suggesting that NV proteinase is a member of the viral cysteine proteinase family. We propose that the proteinase may play a regulatory role in viral replication.

MeSH terms

  • Cysteine Endopeptidases / metabolism*
  • Kinetics
  • Norwalk virus / physiology*
  • Protease Inhibitors / pharmacology
  • Viral Nonstructural Proteins / metabolism*
  • Viral Proteins / metabolism*
  • Virus Replication

Substances

  • Protease Inhibitors
  • Viral Nonstructural Proteins
  • Viral Proteins
  • Cysteine Endopeptidases
  • 3C proteases