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Hydroxyl radicals do not crosslink a DNA-lysozyme complex.

H Werbin, C J Cheng

    Carcinogenesis
    |December 1, 1985
    PubMed
    Summary
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    Hydroxyl radicals, generated by various methods, did not effectively crosslink DNA to lysozyme. These findings challenge previous proposals regarding the role of hydroxyl radicals in DNA-protein crosslinking.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Chemical Biology

    Background:

    • DNA-protein crosslinking is crucial in various biological processes.
    • Previous studies suggested hydroxyl radicals mediate N-acetoxy-2-acetylaminofluorene-induced DNA-lysozyme crosslinking.

    Purpose of the Study:

    • To investigate the role of hydroxyl radicals in DNA-protein crosslinking.
    • To determine if hydroxyl radicals induce crosslinking between lysozyme and DNA (Escherichia coli DNA or pBR322 DNA).

    Main Methods:

    • Utilized systems generating nanomolar hydroxyl radicals: xanthine oxidase-catalyzed xanthine oxidation and iron-catalyzed ascorbic acid oxidation.
    • Employed micromolar hydroxyl radical generation via adenosine nucleotides with ferrous iron and hydrogen peroxide.
    • Quantified radical levels using formaldehyde colorimetric analysis from dimethyl sulfoxide oxidation.

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  • Generated radicals via pulse radiolysis in a nitrous oxide atmosphere.

    • Main Results:

    • No effective crosslinking of lysozyme-DNA complexes was observed with nanomolar hydroxyl radical generating systems.
    • Micromolar hydroxyl radical concentrations also failed to induce significant crosslinking.
    • Pulse radiolysis with similar radical amounts did not induce crosslinking.

    Conclusions:

    • The study's findings do not support a role for hydroxyl radicals in the N-acetoxy-2-acetylaminofluorene-induced crosslinking of DNA to lysozyme.
    • Hydroxyl radicals are unlikely mediators of this specific DNA-protein crosslinking mechanism.