DNA-purine methylation in hepatic chromatin following exposure to dimethylnitrosamine or methylnitrosourea

The investigations reported in this paper were designed to analyze the patterns of DNA-purine methylation in hepatic chromatin following in vivo exposure to the carcinogenic alkylating agents dimethylnitrosamine (DMN) or methylnitrosourea (MNU). Male Sprague-Dawley rats were exposed to [14C]DMN (8 mumoles, 1.0 microCi per mumole per 100 g) or [3H]MNU (15 mumoles, 10 microCi per mumole per 100 g) via gastric intubation. Hepatic chromatin was fractionated into portions having characteristics of template-active euchromatin (S2) and template-repressed heterochromatin (P2) by digestion with DNase II followed by MgCl2 precipitation. Specific DNA purines were identified at 24 hr post-intubation using an isocratic high pressure liquid chromatographic system. A qualitatively similar pattern of 7-methylguanine, O6-methylguanine, 1-methyladenine and 3-methyladenine alkylation was observed in DNA from total chromatin versus heterochromatin at 24 hr following exposure to either carcinogen. These assessments were made at times following carcinogen exposure which produced maximal quantitative differences in alkylation of euchromatin versus heterochromatin DNA. Similar patterns of DNA purine alkylation were observed in total chromatin and heterochromatin. These observations suggest that, once the reactive species is generated and access to chromatin DNA occurs, a similar pattern of DNA-purine alkylation is produced in different regions of the genome.