Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1981 Jul 10;9(13):3089–3103. doi: 10.1093/nar/9.13.3089

A system in mouse liver for the repair of O6-methylguanine lesions in methylated DNA.

J M Bogden, A Eastman, E Bresnick
PMCID: PMC327333  PMID: 7279663

Abstract

An activity from mouse liver with catalyzes the disappearance of O6-methylguanine from DNA methylated with methylnitrosourea has been partially purified by ammonium sulfate fractionation and DNA-cellulose chromatography. The activity does not require divalent metal ions and is not affected by EDTA. It is specific for the repair of O6-methylguanine lesions and does not affect the removal of 7-methylguanine, 7-methyladenine or 3-methyladenine. The disappearance of O6-methylguanine is linear with respect to the concentration of protein and is dependent on incubation temperature. The kinetics and substrate dependence experiments suggest that the protein factor is product-inactivated. Amino acid analysis of hydrolysates of protein obtained after incubation of methylated DNA with the protein factor indicates the presence of radiolabeled S-methyl-L-cysteine, suggesting that during the repair of O6-methylguanine from methylated DNA, the methyl group is transferred to a sulfhydryl of a cysteine residue of a protein. This represents the first such demonstration in a mammalian system.

Full text

PDF
3093

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abanobi S. E., Columbano A., Mulivor R. A., Rajalakshmi S., Sarma D. S. In vivo replication of hepatic deoxyribonucleic acid of rats treated with dimethylnitrosamine: presence of dimethylnitrosamine-induced O6-methylguanine, N7-methylguanine, and N3-methyladenine in the replicated hybrid deoxyribonucleic acid. Biochemistry. 1980 Apr 1;19(7):1382–1387. doi: 10.1021/bi00548a018. [DOI] [PubMed] [Google Scholar]
  2. Abbott P. J., Saffhill R. DNA synthesis with methylated poly(dC-dG) templates. Evidence for a competitive nature to miscoding by O(6)-methylguanine. Biochim Biophys Acta. 1979 Mar 28;562(1):51–61. doi: 10.1016/0005-2787(79)90125-4. [DOI] [PubMed] [Google Scholar]
  3. Beranek D. T., Weis C. C., Swenson D. H. A comprehensive quantitative analysis of methylated and ethylated DNA using high pressure liquid chromatography. Carcinogenesis. 1980 Jul;1(7):595–606. doi: 10.1093/carcin/1.7.595. [DOI] [PubMed] [Google Scholar]
  4. Cox R., Irving C. C. O6-methylguanine accumulates in DNA of mammary glands after administration of N-methyl-N-nitrosourea to rats. Cancer Lett. 1979 Apr;6(4-5):273–278. doi: 10.1016/s0304-3835(79)80045-2. [DOI] [PubMed] [Google Scholar]
  5. Frei J. V., Lawley P. D. Methylation of DNA in various organs of C57B1 mice by a carcinogenic dose of N-methyl-N-nitrosourea and stabiltty of some methylation products up to 18 hours. Chem Biol Interact. 1975 Jun;10(6):413–427. doi: 10.1016/0009-2797(75)90072-1. [DOI] [PubMed] [Google Scholar]
  6. Frei J. V., Swenson D. H., Warren W., Lawley P. D. Alkylation of deoxyribonucleic acid in vivo in various organs of C57BL mice by the carcinogens N-methyl-N-nitrosourea, N-ethyl-N-nitrosourea and ethyl methanesulphonate in relation to induction of thymic lymphoma. Some applications of high-pressure liquid chromatography. Biochem J. 1978 Sep 15;174(3):1031–1044. doi: 10.1042/bj1741031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gerchman L. L., Ludlum D. B. The properties of O 6 -methylguanine in templates for RNA polymerase. Biochim Biophys Acta. 1973 May 10;308(2):310–316. doi: 10.1016/0005-2787(73)90160-3. [DOI] [PubMed] [Google Scholar]
  8. Goth R., Rajewsky M. F. Molecular and cellular mechanisms associated with pulse-carcinogenesis in the rat nerbous system by ethyinitrosourea: ethylation of nucleic acids and elimination rates of ethylated bases from the DNA of different tissues. Z Krebsforsch Klin Onkol Cancer Res Clin Oncol. 1974;82(1):37–64. doi: 10.1007/BF00304382. [DOI] [PubMed] [Google Scholar]
  9. Hartwick R. A., Brown P. R. Evaluation of microparticle chemically bonded reversed-phase packings in the high-pressure liquid chromatographic analysis of nucleosides and their bases. J Chromatogr. 1976 Nov 3;126:679–691. doi: 10.1016/s0021-9673(01)84111-x. [DOI] [PubMed] [Google Scholar]
  10. Karran P., Lindahl T., Griffin B. Adaptive response to alkylating agents involves alteration in situ of O6-methylguanine residues in DNA. Nature. 1979 Jul 5;280(5717):76–77. doi: 10.1038/280076a0. [DOI] [PubMed] [Google Scholar]
  11. Kleihues P., Doerjer G., Keefer L. K., Rice J. M., Roller P. P., Hodgson R. M. Correlation of DNA methylation by methyl(acetoxymethyl)nitrosamine with organ-specific carcinogenicity in rats. Cancer Res. 1979 Dec;39(12):5136–5140. [PubMed] [Google Scholar]
  12. Kleihues P., Margison G. P. Carcinogenicity of N-methyl-N-nitrosourea: possible role of excision repair of O6-methylguanine from DNA. J Natl Cancer Inst. 1974 Dec;53(6):1839–1841. [PubMed] [Google Scholar]
  13. Kleihues P., Margison G. P. Exhaustion and recovery of repair excision of O6-methylguanine from rat liver DNA. Nature. 1976 Jan 15;259(5539):153–155. doi: 10.1038/259153a0. [DOI] [PubMed] [Google Scholar]
  14. Lawley P. D., Orr D. J. Specific excision of methylation products from DNA of Escherichia coli treated with N-methyl-N'-nitro-N-nitrosoguanidine. Chem Biol Interact. 1970 Aug;2(2):154–157. doi: 10.1016/0009-2797(70)90047-5. [DOI] [PubMed] [Google Scholar]
  15. Lawley P. D., Shah S. A. Methylation of DNA by 3H-14C-methyl-labelled N-methyl-N-nitrosourea--evidence for transfer of the intact methyl group. Chem Biol Interact. 1973 Aug;7(2):115–120. doi: 10.1016/0009-2797(73)90020-3. [DOI] [PubMed] [Google Scholar]
  16. Lawley P. D. Some chemical aspects of dose-response relationships in alkylation mutagenesis. Mutat Res. 1974 Jun;23(3):283–295. doi: 10.1016/0027-5107(74)90102-x. [DOI] [PubMed] [Google Scholar]
  17. Lawley P. D., Warren W. Specific excision of ethylated purines from DNA of Escherichia coli treated with N-ethyl-N-nitrosourea. Chem Biol Interact. 1975 Jul;11(1):55–57. doi: 10.1016/0009-2797(75)90066-6. [DOI] [PubMed] [Google Scholar]
  18. Loveless A. Possible relevance of O-6 alkylation of deoxyguanosine to the mutagenicity and carcinogenicity of nitrosamines and nitrosamides. Nature. 1969 Jul 12;223(5202):206–207. doi: 10.1038/223206a0. [DOI] [PubMed] [Google Scholar]
  19. Montesano R., Brésil H., Planche-Martel G., Margison G. P., Pegg A. E. Effect of chronic treatment of rats with dimethylnitrosamine on the removal of O6-methylguanine from DNA. Cancer Res. 1980 Feb;40(2):452–458. [PubMed] [Google Scholar]
  20. Nicoll J. W., Swann P. F., Pegg A. E. Effect of dimethylnitrosamine on persistence of methylated guanines in rat liver and kidney DNA. Nature. 1975 Mar 20;254(5497):261–262. doi: 10.1038/254261a0. [DOI] [PubMed] [Google Scholar]
  21. Olsson M., Lindahl T. Repair of alkylated DNA in Escherichia coli. Methyl group transfer from O6-methylguanine to a protein cysteine residue. J Biol Chem. 1980 Nov 25;255(22):10569–10571. [PubMed] [Google Scholar]
  22. Pegg A. E., Balog B. Formation and subsequent excision of O6-ethylguanine from DNA of rat liver following administration of diethylnitrosamine. Cancer Res. 1979 Dec;39(12):5003–5009. [PubMed] [Google Scholar]
  23. Pegg A. E. Dimethylnitrosamine inhibits enzymatic removal of O6-methylguanine from DNA. Nature. 1978 Jul 13;274(5667):182–184. doi: 10.1038/274182a0. [DOI] [PubMed] [Google Scholar]
  24. Pegg A. E. Enzymatic removal of O6-methylguanine from DNA by mammalian cell extracts. Biochem Biophys Res Commun. 1978 Sep 14;84(1):166–173. doi: 10.1016/0006-291x(78)90278-4. [DOI] [PubMed] [Google Scholar]
  25. Pegg A. E. Formation and metabolism of alkylated nucleosides: possible role in carcinogenesis by nitroso compounds and alkylating agents. Adv Cancer Res. 1977;25:195–269. doi: 10.1016/s0065-230x(08)60635-1. [DOI] [PubMed] [Google Scholar]
  26. Pegg A. E., Hui G. Formation and subsequent removal of O6-methylguanine from deoxyribonucleic acid in rat liver and kidney after small doses of dimethylnitrosamine. Biochem J. 1978 Sep 1;173(3):739–748. doi: 10.1042/bj1730739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pegg A. E., Hui G. Removal of methylated purines from rat liver DNA after administration of dimethylnitrosamine. Cancer Res. 1978 Jul;38(7):2011–2017. [PubMed] [Google Scholar]
  28. Renard A., Verly W. G. A chromatin factor in rat liver which destroys O6-ethylguanine in DNA. FEBS Lett. 1980 May 19;114(1):98–102. doi: 10.1016/0014-5793(80)80868-4. [DOI] [PubMed] [Google Scholar]
  29. Robins P., Cairns J. Quantitation of the adaptive response to alkylating agents. Nature. 1979 Jul 5;280(5717):74–76. doi: 10.1038/280074a0. [DOI] [PubMed] [Google Scholar]
  30. Samson L., Cairns J. A new pathway for DNA repair in Escherichia coli. Nature. 1977 May 19;267(5608):281–283. doi: 10.1038/267281a0. [DOI] [PubMed] [Google Scholar]
  31. Scherer E., Steward A. P., Emmelot P. Kinetics of formation of O6-ethylguanine in, and its removal from liver DNA of rats receiving diethylnitrosamine. Chem Biol Interact. 1977 Oct;19(1):1–11. doi: 10.1016/0009-2797(77)90038-2. [DOI] [PubMed] [Google Scholar]
  32. Singer B. All oxygens in nucleic acids react with carcinogenic ethylating agents. Nature. 1976 Nov 25;264(5584):333–339. doi: 10.1038/264333a0. [DOI] [PubMed] [Google Scholar]
  33. Singer B. N-nitroso alkylating agents: formation and persistence of alkyl derivatives in mammalian nucleic acids as contributing factors in carcinogenesis. J Natl Cancer Inst. 1979 Jun;62(6):1329–1339. [PubMed] [Google Scholar]
  34. Sklar R., Strauss B. Role of the uvrE gene product and of inducible O6-methylguanine removal in the induction of mutations by N-methyl-N'-nitro-N-nitrosoguanidine in Escherichia coli. J Mol Biol. 1980 Nov 15;143(4):343–362. doi: 10.1016/0022-2836(80)90217-x. [DOI] [PubMed] [Google Scholar]
  35. Swann P. F., Mace R. Changes in O6-methylguanine disappearance from rat liver DNA during chronic dimethylnitrosamine administration. A possible similarity between the system removing O6-methylguanine from DNA in rat liver and in Escherichia coli adapted to N-methyl-N'-nitro-N-nitrosoguanidine. Chem Biol Interact. 1980 Aug;31(2):239–245. doi: 10.1016/0009-2797(80)90012-5. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES