Programmed ribosomal frameshifting in decoding the SARS-CoV genome

Virology. 2005 Feb 20;332(2):498-510. doi: 10.1016/j.virol.2004.11.038.

Abstract

Programmed ribosomal frameshifting is an essential mechanism used for the expression of orf1b in coronaviruses. Comparative analysis of the frameshift region reveals a universal shift site U_UUA_AAC, followed by a predicted downstream RNA structure in the form of either a pseudoknot or kissing stem loops. Frameshifting in SARS-CoV has been characterized in cultured mammalian cells using a dual luciferase reporter system and mass spectrometry. Mutagenic analysis of the SARS-CoV shift site and mass spectrometry of an affinity tagged frameshift product confirmed tandem tRNA slippage on the sequence U_UUA_AAC. Analysis of the downstream pseudoknot stimulator of frameshifting in SARS-CoV shows that a proposed RNA secondary structure in loop II and two unpaired nucleotides at the stem I-stem II junction in SARS-CoV are important for frameshift stimulation. These results demonstrate key sequences required for efficient frameshifting, and the utility of mass spectrometry to study ribosomal frameshifting.

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

  • Base Sequence
  • Cell Line
  • Coronavirus / genetics*
  • Frameshifting, Ribosomal / genetics*
  • Genes, Reporter
  • Genome, Viral*
  • Humans
  • Luciferases / genetics
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA, Viral / chemistry
  • RNA, Viral / genetics
  • SARS Virus / genetics*
  • Sequence Alignment
  • Spectrometry, Mass, Electrospray Ionization

Substances

  • RNA, Viral
  • Luciferases