Quantitation of O6-methylguanine-DNA methyltransferase in HeLa cells

R. S. Foote, B. C. Pal, S. Mitra

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Abstract

A synthetic DNA polymer containing [8-3H]O6-methylguanine m6G) was used as a substrate to assay the in situ demethylation of the alkylated base by an activity in HeLa cell extracts. The repair activity appears to be similar to the O6-methylguanine-DNA methyltransferase of E. coli and to be inactivated by reaction with the substrate. Extracts of a methylation-repair proficient (Mer+) cell strain, HeLa CCL2, were found to contain m6G repair activity equivalent to approx. 100 000 molecules of methyltransferase per cell, assuming that each molecule can demethylate one m6G residue. No activity could be detected in the extract of a repair deficient (Mer-) cell strain, HeLa S3, and there is no evidence of an inhibitor of repair activity in this strain.

Original languageEnglish (US)
Pages (from-to)221-228
Number of pages8
JournalMutation Research Letters
Volume119
Issue number3-4
DOIs
StatePublished - Mar 1983

ASJC Scopus subject areas

  • Medicine(all)

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