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Related Experiment Videos

Reverse transcriptase and substrate dependence of the RNA hypermutagenesis reaction

M A Martínez1, M Sala, J P Vartanian

  • 1Unité de Rétrovirologie Moléculaire, Institut Pasteur, Paris, France.

Nucleic Acids Research
|July 25, 1995
PubMed
Summary

Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase is highly sensitive to biased nucleotide concentrations, promoting G-to-A hypermutation. This may explain the high adenine content in lentiviral genomes.

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Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • G-to-A hypermutation occurs during retroviral reverse transcription when deoxyribonucleotide triphosphate (dNTP) concentrations are highly biased.
  • Reverse transcriptases (RTases) from different retroviruses exhibit varying sensitivities to these dNTP biases.

Purpose of the Study:

  • To investigate the sensitivity of different retroviral reverse transcriptases (HIV-1, AMV, MoMLV) to biased dNTP concentrations, specifically focusing on G-to-A hypermutation.
  • To determine the factors influencing HIV-1 RTase's fidelity and its role in genomic composition.

Main Methods:

  • Comparative analysis of reverse transcriptase activity from HIV-1, AMV, and MoMLV.
  • Assessment of enzyme sensitivity to varying deoxycytidine triphosphate (dCTP) to deoxythymidine triphosphate (dTTP) ratios.

Related Experiment Videos

  • Evaluation of discrimination against dUTP and dITP, and insensitivity to pH.

    • Main Results:

    • HIV-1 reverse transcriptase demonstrated the highest sensitivity to biased [dCTP]/[dTTP] ratios, leading to G-to-A hypermutation.
    • HIV-1 RTase effectively discriminated between dUTP, dITP, and DNA precursors, and was not affected by pH.
    • Observed G-to-A hypermutation and multiple strand transfer events were linked to HIV-1 RTase activity.

    Conclusions:

    • The pronounced G-to-A hypermutation propensity of HIV-1 RTase suggests a significant role in shaping lentiviral genome composition.
    • This phenomenon may contribute to the unusually high adenine content observed in HIV-1 and other lentiviral genomes.