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Error-prone polymerization by HIV-1 reverse transcriptase. Contribution of template-primer misalignment, miscoding,

K Bebenek1, J Abbotts, S H Wilson

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

The Journal of Biological Chemistry
|May 15, 1993
PubMed
Summary

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Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase makes errors in homopolymeric sequences. This study investigates models for these errors, finding template-primer misalignment causes deletions and dislocation/miscoding cause substitutions.

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase exhibits error-prone DNA synthesis at homopolymeric sequences.
  • Previous work identified single-nucleotide substitution, addition, and deletion errors, along with processive synthesis termination at these sites.

Purpose of the Study:

  • To test models explaining errors at homopolymeric "hot spots" during DNA synthesis by HIV-1 reverse transcriptase.
  • Investigate the roles of template-primer misalignment, dislocation, and direct miscoding in polymerase errors.

Main Methods:

  • Introduced single-nucleotide changes within and flanking homopolymeric sequences.
  • Analyzed the impact of these sequence modifications on HIV-1 reverse transcriptase error rate, specificity, and termination probability.

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Main Results:

  • Deletion errors in homopolymers are attributed to template-primer misalignment.
  • Base substitution errors result from both direct miscoding and template-primer dislocation.
  • Frameshift error rates and termination probability are influenced by flanking sequences, even distant ones.

Conclusions:

  • The findings elucidate mechanisms underlying HIV-1 reverse transcriptase errors at homopolymeric sequences.
  • A correlation exists between frameshift fidelity and the processivity of DNA synthesis by HIV-1 reverse transcriptase.