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Related Experiment Videos

HIV genetic variation is directed and restricted by DNA precursor availability

J P Vartanian1, U Plikat, M Henry

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

Journal of Molecular Biology
|July 11, 1997
PubMed
Summary

Deoxynucleoside triphosphate (dNTP) imbalances can cause human immunodeficiency virus type 1 (HIV-1) to produce hypermutants. Host cell restrictions and specific dNTP distortions influence viral mutation patterns.

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Deoxynucleoside triphosphate (dNTP) pools are crucial for DNA replication fidelity.
  • Imbalances in dNTP concentrations are known to introduce errors during DNA synthesis.
  • The fidelity of human immunodeficiency virus type 1 (HIV-1) replication is essential for its lifecycle and evolution.

Purpose of the Study:

  • To investigate the impact of dNTP imbalances on HIV-1 replication fidelity.
  • To characterize the types of mutations (hypermutants) produced under dNTP biased conditions.
  • To compare the in vitro generated mutant spectrum with naturally occurring HIV-1 hypermutants.

Main Methods:

  • Using detergent-permeabilized HIV-1 virions with controlled, biased dNTP concentrations.

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  • Employing a genetic screen utilizing the blue/white beta-galactosidase complementation assay.
  • Analyzing mutations in HIV-1 recovered from infected U937 cells and primary peripheral blood mononuclear cells (PBMCs).

    • Main Results:

    • Biased dNTP concentrations readily produced various hypermutants in vitro.
    • The spectrum of induced hypermutants differed from naturally occurring ones, suggesting host cell influence.
    • G --> A hypermutants were specifically recovered from thymidine-treated HIV-infected U937 cells.
    • A small percentage (1-2%) of primary cells (resting or activated PBMCs) yielded hypermutants, indicating endogenous dNTP distortions.

    Conclusions:

    • dNTP imbalances can significantly affect HIV-1 replication fidelity, leading to hypermutation.
    • Host cellular mechanisms may modulate or restrict the spectrum of viral mutations.
    • Endogenous dNTP pool distortions in a fraction of primary cells could contribute to somatic and germline mutations, influencing mammalian and viral genome evolution.