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The accuracy of reverse transcriptase from HIV-1.

A study was conducted to determine the fidelity of DNA synthesis catalyzed in vitro by the reverse transcriptase from a human immunodeficiency virus type 1 (HIV-1). Like other retroviral reverse transcriptases, the HIV-1 enzyme does not correct errors by exonucleolytic proofreading. Measurements with M13mp2-based fidelity assays indicated that the HIV-1 enzyme, isolated either from virus particles or from Escherichia coli cells infected with a plasmid expressing the cloned gene, was exceptionally inaccurate, having an average error rate per detectable nucleotide incorporated of 1/1700. It was, in fact, the least accurate reverse transcriptase described to date, one-tenth as accurate as the polymerases isolated from avian myeloblastosis or murine leukemia viruses, which have average error rates of approximately 1/17,000 and approximately 1/30,000, respectively. DNA sequence analyses of mutations generated by HIV-1 polymerase showed that base substitution, addition, and deletion errors were all produced. Certain template positions were mutational hotspots where the error rate could be as high as 1 per 70 polymerized nucleotides. The data are consistent with the notion that the exceptional diversity of the HIV-1 genome results from error-prone reverse transcription.

Pubmed ID: 2460925

Authors

  • Roberts JD
  • Bebenek K
  • Kunkel TA

Journal

Science (New York, N.Y.)

Publication Data

November 25, 1988

Associated Grants

None

Mesh Terms

  • Avian myeloblastosis virus
  • DNA
  • DNA-Directed DNA Polymerase
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Exonucleases
  • HIV
  • Moloney murine leukemia virus
  • Mutation
  • Nucleotides
  • RNA-Directed DNA Polymerase
  • Recombinant Proteins