Formation and removal of DNA adducts after treatment of Chinese hamster ovary cells with N-methyl- and N-ethyl-N-nitrosourea.
We have studied formation and stability of alkylguanines following treatment of Chinese hamster ovary cells with either N-[3H]methyl-N-nitrosourea (MeNOUr) (applied at 50 microM and 40 microM concentrations) or N-[3H]ethyl-N-nitrosourea (EtNOUr) (applied at 43.1 microM). Analyses of acid hydrolysates of the methylated DNA revealed that 9.3% and 57.0% of the total DNA were O6-methylguanine (m6Gua) and 7-methylguanine (m7Gua), respectively. Analysis of enzymic hydrolysate resulted in 8.2% m6Gua and 50.3% m7Gua. For ethylation, the % of ethylated purines identified as O6-ethylguanine (e6Gua) and 7-ethylguanine (e7Gua) were 20.4% and 31.3%, respectively. Half-lives of the main alkylated purines were determined by analysing DNA of dividing cultures over a time interval of 48 h after treatment with carcinogens. Half-lives measured for methylated DNA bases were: m1Ade, 20.6 h; m3Ade, 25.5 h; m7Ade, 0.9 h; m3Gua, 1.1 h; m6Gua, infinity; m7Gua, 39.1 h. Determinations at the level of deoxyribonucleosides resulted in similar half-lives: m3dA, 15.2 h; m7dA, 2.7 h; m3dG, 2.3 h; m6dG, 224 h; m7dG, 25.6 h. The corresponding values for ethylated purines were: e3Ade, 2.9 h; e7Ade, 7.1 h; e3Gua, 1.4 h; e6Gua, infinity; e7Gua, 42.6 h. The relatively high yields of the premutagenic m6Gua and e6Gua, and their long half-lives (greater than or equal to 224 h) are consistent with the suggestion that these adducts play a dominant role in mutation induction at the hypoxanthine-guanine phosphoribosyltransferase (hgprt) locus in CHO cells.
Thielmann
H W
HW
Institute of Biochemistry, German Cancer Research Center, Heidelberg, F.R.G.
Schröder
C H
CH
Hsie
A W
AW
D000478:Alkylation; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D004252:DNA Mutational Analysis; D004260:DNA Repair; D005038:Ethylnitrosourea; D008745:Methylation; D008770:Methylnitrosourea; D013997:Time Factors
D008770:Methylnitrosourea; D004247:DNA; D005038:Ethylnitrosourea