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  • Title: Reconstitution of intercalator-induced DNA scission by an active component from nuclear extracts.
    Author: Filipski J, Yin J, Kohn KW.
    Journal: Biochim Biophys Acta; 1983 Oct 13; 741(1):116-22. PubMed ID: 6688534.
    Abstract:
    Treatment with intercalating agents causes formation of protein-associated DNA breaks in mammalian cells in culture and in the nuclei isolated from these cells. We found that this effect, when induced by the intercalator m-AMSA, required a component which could be dissociated from nuclei by 0.3 M NaCl. The effect was restored by combining the extracted nuclei with the nuclear extract. The active component of the extract eluted in gel filtration at a point corresponding to a molecular weight of 800 000. During its reaction with DNA, DNA-protein links and DNA breaks appeared in approximately equal frequencies. In this respect the reaction stimulated by m-AMSA resembled the reaction of a topoisomerase with DNA. However, intercalator-stimulated formation of protein-associated DNA breaks differed from the activity of the nuclear topoisomerase I in that there was a different optimum salt concentration and a different apparent molecular weight.
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