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Journal of virology1991; 65(4); 1952-1959; doi: 10.1128/JVI.65.4.1952-1959.1991

Equine infectious anemia virus and human immunodeficiency virus DNA synthesis in vitro: characterization of the endogenous reverse transcriptase reaction.

Abstract: The endogenous reverse transcriptase reaction of equine infectious anemia virus (EIAV) has been studied, and conditions allowing synthesis of full-length minus-strand DNA have been determined. In contrast to results reported for other retroviruses, synthesis of EIAV full-length minus-strand DNA was not impaired by high concentrations of Nonidet P-40, a nonionic detergent used to make the virion envelope permeable. All components of the reaction were titrated for maximum synthesis of complete minus strands, and a time course under the standardized conditions was determined. Minor subgenomic bands were observed in some cases, and both the size and proportion varied with reaction conditions. Conditions established for full-length EIAV DNA synthesis also allowed full-genome-length human immunodeficiency virus type 1 DNA synthesis. The human immunodeficiency virus type 1 DNA product contained a greater proportion of reverse transcripts that were shorter than the complete virus genome. Also in contrast to EIAV, the endogenous synthesis of high-molecular-weight human immunodeficiency virus type 1 DNA was drastically reduced at Nonidet P-40 concentrations above 0.02%. These results indicated that a detergent-stable core is not a property shared by all lentiviruses. The EIAV virion synthetic machinery is unusually stable and provides a convenient system for further in vitro study of reverse transcription.
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Publication Date: 1991-04-01 PubMed ID: 1705993PubMed Central: PMC240025DOI: 10.1128/JVI.65.4.1952-1959.1991Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates the synthesis process of equine infectious anemia virus (EIAV) and human immunodeficiency virus (HIV) DNA. The study finds the EIAV DNA synthesis reaction conditions that allow for full-length minus-strand DNA, and observes its stability in the presence of high concentrations of a detergent, Nonidet P-40. The methods also enabled the full-genome-length HIV-1 DNA synthesis, however the HIV DNA product contained a higher proportion of shorter reverse transcripts, and its synthesis was significantly reduced when Nonidet P-40 concentrations exceeded 0.02%.

EIAV and HIV DNA Synthesis

  • The research focused on the endogenous reverse transcriptase reaction of the EIAV, aiming to characterize the conditions that enable synthesis of full-length minus-strand DNA.
  • Different from other retroviruses, EIAV full-length minus-strand DNA synthesis was not affected by high concentrations of a nonionic detergent called Nonidet P-40. This detergent is used to permeabilize the virion envelope and its tolerance indicates EIAV’s unusual stability.

Components of the Reaction

  • The researchers optimized all components of the reaction for maximum synthesis of complete minus strands and determined a time course under these standardized conditions.
  • Notably, minor subgenomic bands were observed in certain cases, and both their size and proportion varied depending on the reaction conditions.

Comparative Synthesis in HIV

  • Using the same conditions established for EIAV DNA synthesis, researchers were also able to facilitate full-genome-length HIV type 1 DNA synthesis.
  • However, the HIV type 1 DNA product contained more reverse transcripts that were shorter than the complete virus genome, compared to EIAV.
  • The reaction to Nonidet P-40 was drastically different in HIV, where the endogenous synthesis of high-molecular-weight HIV type 1 DNA was significantly reduced at concentrations above 0.02%.
  • These findings suggested that a detergent-stable core is not a consistent property among all lentiviruses.

Implications for Further Research

  • As a result of its stability and predictable reactions, the EIAV virion synthetic machinery provides a convenient system for further in vitro study of reverse transcription.
  • The uncovering of these properties may aid in the development of treatments or interventions for equine and human diseases caused by these retroviruses.

Cite This Article

APA
Borroto-Esoda K, Boone LR. (1991). Equine infectious anemia virus and human immunodeficiency virus DNA synthesis in vitro: characterization of the endogenous reverse transcriptase reaction. J Virol, 65(4), 1952-1959. https://doi.org/10.1128/JVI.65.4.1952-1959.1991

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 65
Issue: 4
Pages: 1952-1959

Researcher Affiliations

Borroto-Esoda, K
  • Division of Virology, Wellcome Research Laboratories, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.
Boone, L R

    MeSH Terms

    • Acetates / metabolism
    • Acetic Acid
    • Animals
    • Base Sequence
    • Cell Membrane Permeability
    • Cells, Cultured
    • DNA Replication / drug effects
    • DNA, Viral / biosynthesis
    • HIV-1 / enzymology
    • HIV-1 / genetics
    • HIV-1 / growth & development
    • Horses
    • In Vitro Techniques
    • Infectious Anemia Virus, Equine / enzymology
    • Infectious Anemia Virus, Equine / genetics
    • Infectious Anemia Virus, Equine / growth & development
    • Molecular Sequence Data
    • Nucleotides / metabolism
    • Octoxynol
    • Polyethylene Glycols / pharmacology
    • RNA-Directed DNA Polymerase / metabolism
    • Virion / drug effects
    • Virion / genetics
    • Virion / growth & development
    • Virus Replication / drug effects

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