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Related Experiment Videos

DNA methyltransferase induced by frog virus 3.

D B Willis, R Goorha, A Granoff

    Journal of Virology
    |January 1, 1984
    PubMed
    Summary

    Frog virus 3 infection induces a novel cytoplasmic DNA methyltransferase. This virus-encoded enzyme methylates cytosine bases in viral DNA, preferring double-stranded and unmethylated DNA substrates.

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    Area of Science:

    • Virology
    • Molecular Biology
    • Epigenetics

    Background:

    • Frog virus 3 (FV3) exhibits extensive DNA methylation (>20% of cytosines).
    • The origin of this high methylation level, whether host or viral, was previously unclear.

    Purpose of the Study:

    • To investigate if FV3 encodes a specific DNA methyltransferase responsible for its DNA methylation.
    • To characterize the properties and kinetics of this potential viral enzyme.

    Main Methods:

    • Analysis of DNA methyltransferase activity in FV3-infected fathead minnow cells.
    • Characterization of enzyme kinetics, substrate specificity (single-stranded vs. double-stranded DNA, methylated vs. unmethylated DNA), and optimal exogenous substrate (dCpdG).
    • Comparison of enzyme activity in infected cells versus uninfected cells and analysis of a 5-azacytidine-resistant FV3 mutant with unmethylated DNA.

    Main Results:

    • A novel cytoplasmic DNA methyltransferase activity was detected 3 hours post-infection, independent of viral DNA replication.
    • The cytoplasmic enzyme demonstrated a preference for double-stranded, unmethylated DNA and utilized dCpdG as an exogenous substrate.
    • A FV3 mutant lacking DNA methylation failed to induce this cytoplasmic methyltransferase activity.

    Conclusions:

    • The observed DNA methyltransferase activity is likely encoded by frog virus 3.
    • This virus-specific enzyme contributes to the high level of cytosine methylation in FV3 DNA.

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