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The accuracy of reverse transcriptase from HIV-1.

J D Roberts1, K Bebenek, T A Kunkel

  • 1Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.

Science (New York, N.Y.)
|November 25, 1988
PubMed
Summary

The human immunodeficiency virus type 1 (HIV-1) reverse transcriptase is highly inaccurate, introducing errors during DNA synthesis. This error-prone process contributes to the genetic diversity observed in the HIV-1 genome.

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Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Retroviral reverse transcriptases play a crucial role in viral replication.
  • Proofreading mechanisms are essential for maintaining DNA synthesis fidelity in many polymerases.

Purpose of the Study:

  • To assess the DNA synthesis fidelity of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase in vitro.
  • To compare the accuracy of HIV-1 reverse transcriptase with other retroviral polymerases.

Main Methods:

  • Utilized M13mp2-based fidelity assays to quantify error rates.
  • Analyzed DNA sequences of mutations generated by the HIV-1 polymerase.
  • Isolated HIV-1 reverse transcriptase from both viral particles and recombinant Escherichia coli.

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Main Results:

  • HIV-1 reverse transcriptase exhibits exceptionally low fidelity, with an average error rate of 1/1700 nucleotides.
  • The enzyme lacks exonucleolytic proofreading activity, unlike some other polymerases.
  • Generated mutations included base substitutions, additions, and deletions, with hotspots showing error rates as high as 1/70.

Conclusions:

  • HIV-1 reverse transcriptase is the least accurate retroviral reverse transcriptase characterized to date.
  • The high error rate of HIV-1 reverse transcriptase likely contributes to the genetic diversity and rapid evolution of the HIV-1 genome.