KINETICS OF O6-METHYLGUANINE REPAIR IN HUMAN NORMAL AND ATAXIA TELANGIECTASIA CELL-LINES AND CORRELATION OF REPAIR CAPACITY WITH CELLULAR-SENSITIVITY TO METHYLATING AGENTS

Abstract

Human lymphoblastoid cell lines from normal individuals and from patients with ataxia telangiectasia were either proficient or deficient in their ability to repair the mutagenic DNA adduct O6-methylguanine that is induced by methylating carcinogens. There was no relationship between the capacity to repair O6-methylguanine and the ataxia telangiectasia phenotype. Time-course studies done following a short pulse (2 min) of alkylation with 0.5 .mu.g of N-[3H]methyl-N''-nitro-N-nitrosoguanidine per ml revealed that the repair of O6-methylguanine in human lymphoblastoid lines proficient in this ability is a rapid process which proceeds with a t1/2 of 10-15 min. Lymphoblastoid lines with deficient capacity to repair this DNA adduct were hypersensitive to the cytotoxic effect of the methylating carcinogens N-methyl-N''-nitro-N-nitrosoguanidine, N-methyl-N-nitrosourea and methyl methanesulfonate, and this hypersensitivity was correlated with the relative amount of O6-methylguanine induced by each of the 3 chemicals. This was taken as an indication of the lethality of unrepaired O6-methylguanine. The extent of DNA repair synthesis induced by the 3 carcinogens was the same in cell lines proficient and deficient in O6-methylguanine repair, indicating no major deficiency in an excision repair pathway in the hypersensitive cell lines.