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Related Experiment Videos

Effect of dimethyl sulfoxide on reverse transcriptase activity

M B Jucá1, H Aoyama

  • 1Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas, SP, Brasil.

Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Medicas E Biologicas
|March 1, 1995
PubMed
Summary

Dimethyl sulfoxide (DMSO) enhances avian myeloblastosis virus reverse transcriptase activity at low concentrations, potentially by reducing the Km for poly(rA). Higher DMSO concentrations inhibit this enzyme activity.

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Area of Science:

  • Biochemistry
  • Molecular Biology

Background:

  • Dimethyl sulfoxide (DMSO) is a common solvent with known biological effects.
  • Reverse transcriptase enzymes are crucial in retroviral replication and molecular biology.
  • Understanding enzyme kinetics and the influence of chemical agents is vital for research.

Purpose of the Study:

  • To investigate the effect of dimethyl sulfoxide (DMSO) on avian myeloblastosis virus (AMV) reverse transcriptase activity.
  • To determine the concentration-dependent effects of DMSO on enzyme kinetics.
  • To explore the potential mechanisms behind DMSO's influence on AMV reverse transcriptase.

Main Methods:

  • Enzyme activity assays were performed using avian myeloblastosis virus reverse transcriptase.
  • Activated DNA and polyriboadenylic acid (poly(rA)) were used as templates.

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  • Divalent cations (Mg2+ and Mn2+) were employed, and DMSO concentrations were varied.

    • Main Results:

    • Low concentrations of DMSO (around 5%) stimulated AMV reverse transcriptase activity by up to 40%.
    • DMSO also stimulated poly(rA)-dependent activity with Mn2+.
    • Inhibition was observed at DMSO concentrations above 15%, except for a 100% activation with a poly(rA) analog at 20% DMSO.
    • DMSO reduced the apparent Km for poly(rA) from 9.1 to 3.3 µg/ml, suggesting a mechanism for stimulation.

    Conclusions:

    • DMSO exhibits a biphasic effect on AMV reverse transcriptase activity: stimulation at low concentrations and inhibition at high concentrations.
    • The observed stimulation may be attributed to a decrease in the enzyme's affinity for the poly(rA) template.
    • These findings highlight DMSO's complex interaction with reverse transcriptase and have implications for its use in molecular biology applications.