'Primer alignment-and-extension': a novel mechanism of viral RNA recombination responsible for the rescue of inactivated poliovirus cDNA clones

J Gen Virol. 1999 Aug:80 ( Pt 8):1889-1897. doi: 10.1099/0022-1317-80-8-1889.

Abstract

In the course of experiments designed to assess the potential role of alternative open reading frames (ORF) present in the 5'-terminal untranslated region (5'-UTR) of poliovirus type 1 (Mahoney strain) genomic RNA, we came across a double mutation that completely abrogated the infectivity of full-length cDNA clones. The infectivity was rescued in trans by cotransfecting COS-1 cells with short RNA transcripts of the wild-type 5'-UTR of poliovirus type 2 Lansing, provided a free 3'-OH was available. Direct sequencing of the viral RNA revealed that the infectious viruses recovered were recombinants Lansing/Mahoney, with variable points of 'crossing-over'. A novel mechanism of RNA-RNA recombination, which we propose to call 'primer alignment-and-extension', is described that would explain the high rate of recombination of RNA viruses observed in natural conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions
  • Animals
  • Base Sequence
  • COS Cells
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Codon, Initiator
  • DNA, Complementary
  • DNA, Viral
  • Genome, Viral
  • Humans
  • Molecular Sequence Data
  • Mutagenesis
  • Poliovirus / genetics*
  • Protein Biosynthesis
  • RNA
  • RNA, Viral*
  • Recombination, Genetic*
  • Transfection
  • Vero Cells
  • Virion

Substances

  • 5' Untranslated Regions
  • Codon, Initiator
  • DNA, Complementary
  • DNA, Viral
  • RNA primers
  • RNA, Viral
  • RNA