Fate of direct and inverted repeats in the RNA hypermutagenesis reaction

Nucleic Acids Res. 1996 Jan 15;24(2):253-6. doi: 10.1093/nar/24.2.253.

Abstract

RNA hypermutagenesis results from cDNA synthesis in the presence of highly biased dNTP precursor concentrations and preferentially exploits human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Such reaction conditions slow down DNA synthesis, which might be conducive to strand transfer and deletion. This has been investigated. A 6 bp inverted repeat nested between 10 bp repeats was efficiently deleted at dCTP concentrations typically used. Inter- or intramolecular strand transfer between 10 bp repeated sequences separated by runs of templated G residues occurred, but at lower concentrations. If RNA hypermutagenesis of a sequence containing direct and inverted repeats is unavoidable, avian myeloblastosis virus (AMV) reverse transcriptase could be used, as strand transfer occurs with much diminished dCTP substrate dependence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA, Complementary / biosynthesis
  • Deoxycytosine Nucleotides
  • HIV Reverse Transcriptase
  • Humans
  • Molecular Sequence Data
  • Mutagenesis / genetics*
  • Osmolar Concentration
  • RNA / genetics*
  • RNA-Directed DNA Polymerase / metabolism
  • Repetitive Sequences, Nucleic Acid / genetics*
  • Retroviridae / enzymology
  • Sequence Deletion

Substances

  • DNA, Complementary
  • Deoxycytosine Nucleotides
  • RNA
  • HIV Reverse Transcriptase
  • RNA-Directed DNA Polymerase