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Nucleic Acids Research, 1985, Vol. 13, No. 8 2683-2698
© 1985

Articles

Methyl phosphotriesters in alkylated DNA are repaired by the Ada regulatory protein of E. coli

Tommie V. McCarthy and Tomas Lindahl

Imperial Cancer Research Fund, Mill Hill Laboratories London NW7 1AD, UK

Received February 19, 1985. Accepted March 27, 1985.

The E.coli ada+ gene product that controls the adaptive response to alkylating agents has been purified to apparent homogeneity using an overproducing expression vector system. This 39 kDa protein repairs 06-methylguanine and 04-methylthymine residues in alkylated DNA by transfer of the methyl group from the base to a cysteine residue in the protein itself. The Ada protein also corrects one of the stereoisomers of methyl phosphotriesters in DNA by the same mechanism, while the other isomer is left unrepaired. Different cysteine residues in the Ada protein are used as acceptors in the repair of methyl groups derived from phosphotriesters and base residues.


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