Reovirus polymerase lambda 3 localized by cryo-electron microscopy of virions at a resolution of 7.6 A

Nat Struct Biol. 2003 Dec;10(12):1011-8. doi: 10.1038/nsb1009. Epub 2003 Nov 9.

Abstract

Reovirus is an icosahedral, double-stranded (ds) RNA virus that uses viral polymerases packaged within the viral core to transcribe its ten distinct plus-strand RNAs. To localize these polymerases, the structure of the reovirion was refined to a resolution of 7.6 A by cryo-electron microscopy (cryo-EM) and three-dimensional (3D) image reconstruction. X-ray crystal models of reovirus proteins, including polymerase lambda 3, were then fitted into the density map. Each copy of lambda 3 was found anchored to the inner surface of the icosahedral core shell, making major contacts with three molecules of shell protein lambda 1 and overlapping, but not centering on, a five-fold axis. The overlap explains why only one copy of lambda 3 is bound per vertex. lambda 3 is furthermore oriented with its transcript exit channel facing a small channel through the lambda 1 shell, suggesting how the nascent RNA is passed into the large external cavity of the pentameric capping enzyme complex formed by protein lambda 2.

Publication types

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

MeSH terms

  • Cryoelectron Microscopy
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / ultrastructure*
  • Image Processing, Computer-Assisted
  • Nucleic Acid Conformation
  • Protein Conformation
  • RNA, Double-Stranded / ultrastructure*
  • RNA, Viral / ultrastructure
  • RNA-Dependent RNA Polymerase / chemistry
  • RNA-Dependent RNA Polymerase / ultrastructure*
  • Reoviridae / enzymology*
  • Reoviridae / ultrastructure*
  • Sensitivity and Specificity
  • Viral Proteins / chemistry
  • Viral Proteins / ultrastructure*
  • Virion / enzymology
  • Virion / ultrastructure

Substances

  • RNA, Double-Stranded
  • RNA, Viral
  • Viral Proteins
  • lambda3 protein, Reovirus
  • RNA-Dependent RNA Polymerase
  • DNA-Directed RNA Polymerases