Pol of gag-pol fusion protein required for encapsidation of viral RNA of yeast L-A virus

Nature. 1992 Oct 22;359(6397):746-9. doi: 10.1038/359746a0.

Abstract

Double-stranded RNA viruses have an RNA-dependent RNA polymerase activity associated with the viral particles which is indispensable for their replication cycle. Using the yeast L-A double-stranded RNA virus we have investigated the mechanism by which the virus encapsidates its genomic RNA and RNA polymerase. The L-A gag gene encodes the principal viral coat protein and the overlapping pol gene is expressed as a gag-pol fusion protein which is formed by a -1 ribosomal frameshift. Here we show that Gag alone is sufficient for virus particle formation, but that it fails to package the viral single-stranded RNA genome. Encapsidation of the viral RNA requires only a part of the Pol region (the N-terminal quarter), which is presumably distinct from the RNA polymerase domain. Given that the Pol region has single-stranded RNA-binding activity, these results are consistent with our L-A virus encapsidation model: the Pol region of the fusion protein binds specifically to the viral genome (+) strand, and the N-terminal gag-encoded region primes polymerization of Gag to form the capsid, thus ensuring the packaging of both the viral genome and the RNA polymerase.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Capsid / metabolism*
  • DNA Mutational Analysis
  • Gene Products, gag / metabolism*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Oligodeoxyribonucleotides / chemistry
  • RNA Viruses / genetics*
  • RNA, Double-Stranded / genetics*
  • RNA-Dependent RNA Polymerase / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Structure-Activity Relationship
  • Viral Fusion Proteins / metabolism*
  • Virus Replication*

Substances

  • Gene Products, gag
  • Oligodeoxyribonucleotides
  • RNA, Double-Stranded
  • Viral Fusion Proteins
  • RNA-Dependent RNA Polymerase