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Measurement of the removal of O6-methylguanine and O4-methylthymine from oligodeoxynucleotides using an
N M Mironov1, C P Wild, G Martel-Planche
1Unit of Mechanisms of Carcinogenesis, International Agency for Research on Cancer, Lyon, France.
Abstract:
A sensitive and rapid procedure for measurement of alkyltransferase repair activity involving oligodeoxynucleotides followed by immunoprecipitation is described. Dodecadeoxynucleotides containing O6-methylguanine or O4-methylthymine were used as substrates for alkyltransferases and the reaction products of methylated or demethylated substrates were separated by precipitation with highly specific antibodies. This approach for O6-alkylguanine-DNA alkyltransferase measurement is far more rapid than when the reaction products are separated by chromatography. This technique makes the assay applicable to large-scale epidemiological or clinical studies and suggests a similar methodology could be applied for other DNA repair enzymes.
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