Crystal structure and functional analysis of the SARS-coronavirus RNA cap 2'-O-methyltransferase nsp10/nsp16 complex

PLoS Pathog. 2011 May;7(5):e1002059. doi: 10.1371/journal.ppat.1002059. Epub 2011 May 26.

Abstract

Cellular and viral S-adenosylmethionine-dependent methyltransferases are involved in many regulated processes such as metabolism, detoxification, signal transduction, chromatin remodeling, nucleic acid processing, and mRNA capping. The Severe Acute Respiratory Syndrome coronavirus nsp16 protein is a S-adenosylmethionine-dependent (nucleoside-2'-O)-methyltransferase only active in the presence of its activating partner nsp10. We report the nsp10/nsp16 complex structure at 2.0 Å resolution, which shows nsp10 bound to nsp16 through a ∼930 Ų surface area in nsp10. Functional assays identify key residues involved in nsp10/nsp16 association, and in RNA binding or catalysis, the latter likely through a SN2-like mechanism. We present two other crystal structures, the inhibitor Sinefungin bound in the S-adenosylmethionine binding pocket and the tighter complex nsp10(Y96F)/nsp16, providing the first structural insight into the regulation of RNA capping enzymes in +RNA viruses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / analogs & derivatives
  • Adenosine / metabolism
  • Crystallization
  • Magnesium / metabolism
  • Methyltransferases / chemistry*
  • Methyltransferases / metabolism*
  • Mutation / genetics
  • Plasmids
  • Protein Binding
  • RNA Caps / metabolism*
  • RNA, Viral / metabolism*
  • S-Adenosylmethionine / metabolism
  • Severe acute respiratory syndrome-related coronavirus / genetics*
  • Viral Nonstructural Proteins / chemistry*
  • Viral Nonstructural Proteins / metabolism*

Substances

  • Nsp10 protein, SARS virus
  • RNA Caps
  • RNA, Viral
  • Viral Nonstructural Proteins
  • S-Adenosylmethionine
  • Methyltransferases
  • Nsp16 protein, SARS virus
  • RNA 2'-O-methyltransferase
  • Magnesium
  • Adenosine
  • sinefungin