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Nucleic Acids Research, 1985, Vol. 13, No. 6 1939-1952
© 1985

Articles

Cloning of the E. coli O6-methylguanine and methylphosphotriester methyltransferase gene using a functional DNA repair assay

Geoffrey P. Margison1, Donald P. Cooper1,3 and John Brennand2

1Department of Carcinogenesis Manchester, M20 9BX, UK 2Department of Biochemical Genetics, Paterson Laboratories, Christie Hospital Manchester, M20 9BX, UK

Received December 13, 1984. Revised February 25, 1985. Accepted February 25, 1985.

Akylating agents react with various nitrogen and oxygen atoms in DNA and many of the products are substrates repair processes. Oxygen atom derivatives such as 06 -methylguanine (06 -meG) 04 -methylthymine and methylphosphotriesters (MP) have been shown to undergo repair by methyl group removal. The proteins involved in the latter reaction can be considered to be methyltranferases (MT) because their action results in the transfer of the methyl group to a cysteine residue within a polypeptide. A rapid and sensitive assay for MT activity has been developed and used to screen extracts of bacteria harbouring an E.coli genomic DNA library carried in a glasmid vector. We report here the cloning of an E.coli gene coding for 06 -meG and MP MT repair functions. These two activities reside on a 37Kd protein that can undergo a host-dependent cleavage to produce an l8Kd protein which contains only 06 -meG MT and a l3Kd protein which contains only MP MT.

3Present address: Cancer Research Unit, University of York, York YO 5DU. UK


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