DNA Repair of Alkylation Damage and Its Relevance to Mutation Fixation in Mammalian Cells

  • M. Bignami
  • G. Aquilina
  • A. Zijno
  • G. Frosina
  • A. Abbondandolo
  • E. Dogliotti
Chapter

Abstract

Simple monofunctional alkylating agents such as Nmethyl-N’-nitro-N-nitrosoguanidine (MNNG) and N-ethyl-Nnitrosourea (ENU) act directly on oxygen and ring nitrogen atoms of DNA bases, and on oxygen atoms of the phosphate groups to generate a variety of adducts. Two major pathways for the repair of alkylation damage have been identified. The removal of O6-methylguanine (O6-meG) adducts proceeds via the transfer of the alkyl group to a cysteine residue of an acceptor protein, O6-alkylguanine-DNA methyltransferase (MT), which thereby becomes irreversibly inactivated 1,2. Other alkyl adducts are excised through the action of DNA glycosylases which hydrolyze the N-glycosylic bond linking the damaged base to the sugar phosphate backbone. The pathways for the repair of alkylation damage in mammalian cells have not been characterized to the same extent as in bacteria, although the basic strategies employed are the same in prokaryotes and eukaryotes. For example, an alkyltransferase which transfers in a stoichiometric fashion the methyl group from the oxygen 6 of guanine to a cysteine thiol in the protein has been identified3,4 along with a glycosylase activity capable of excising alkyl groups from nitrogen 3 of adenine and guanine and from nitrogen 7 of guanine5. Among the lesions produced by alkylating agents considerable evidence suggests that O6 -alkylguanine (O6 -alkG) is a major premutagenic lesion responsible for carcinogenicity and mutagenicity. Accumulation and persistence of O6 -meG and O6 -ethylguanine (O6 -etG) in target organs for carcinogenesis by alkylating agents has been demonstrated6,7. Comparative mutagenicity studies using a series of human fibroblast cell lines that differ in their levels of MT have shown a low mutagenicity in lines with high capacity to remove O6 -alkG8. In contrast some reports suggest that other lesions may be implicated as well in the mutagenic process9. Good candidates are the Oalkylpyrimidines (O4 -alkylthymine and O2 -alkylthymine)10.

Keywords

Chinese Hamster Ovary Cell Methyl Transferase Cysteine Thiol Methylating Agent Sugar Phosphate Backbone 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Olsson, M. and Lindahl, T. Repair of alkylated DNA in E. coli: methyl group transfer from 06 -methylguanine to a protein cystein residue, J. Biol. Chem., 255: 10569–10571 (1980)PubMedGoogle Scholar
  2. 2.
    Demple, B., Sedgwick, B. Robins, P., Totty, N., Waterfield, M.D. and Lindahl, T. Active site and complete sequence of the suicidal methyltransferase that counters alkylation mutagenesis, Proc. Natl. Acad. Sci. USA, 82: 26882692 (1985).Google Scholar
  3. 3.
    Bogden, J.M., Eastman, A. and Bresnick, E. A system in mouse liver for the repair of 06 -methylguanine lesions in methylated DNA, Nucl. Acids Res., 9: 3089–3103 (1981).CrossRefGoogle Scholar
  4. 4.
    Pegg, A.E., Roberfroid, M., von Bahr, C., Foote, R.S., Mitra, S., Bresil,H., Likhachev, A. and Montesano, R. Removal of 06 -methylguanine from DNA by human liver fractions, Proc. Natl. Acad. Sci. USA, 79: 5162–5165 (1982).CrossRefGoogle Scholar
  5. 5.
    Singer, B. and Brent, T.P. Human lymphoblasts contain DNA glycosylase activity excising N-3 and N-7 methyl and ethyl purines but not 06 -alkylguanines or 1-alkyladenines Proc.Natl.Acad. Sci. 78: 856–860 (1981).Google Scholar
  6. 6.
    Goth, R. and Rajewski, M.F. Persistence of 06–ethylguanine in rat brain DNA: correlation with nervous system-specific carcinogenesis by ethylnitrosourea. Proc. Natl. Acad. Sci. USA 71: 639–643 (1974).PubMedCrossRefGoogle Scholar
  7. Montesano, R., Bresil, H., Planche-Martel, G., Margison, P.G., and Pegg, A.E. Effect of chronic treatment of rats with dimethylnitrosoamine on the removal of 06 -methylguanine from DNA. Cancer Res. 40: 452–458 (1980).Google Scholar
  8. 8.
    Domoradzki, J., Pegg, A.E., Dolan, M.E., Maher, V.M. and McCormick, J.J.Correlation between 06 -methylguanine DNAmethyltransferase activity and resistance of human cells to the cytotoxic and mutagenic effect of N-methyl-N’-nitro-Nnitrosoguanidine, Carcinogenesis, 5: 1641–1647 (1984).PubMedCrossRefGoogle Scholar
  9. 9.
    Fox, M. and Brennand, J. Evidence for the involvement of lesions other than 06 -alkylguanine in mammalian cell mutagenesis, Carcinogenesis, 1: 795–799 (1980)PubMedCrossRefGoogle Scholar
  10. 10.
    Singer, B., Sagi, J. and Kusmierek, J.T. Escherichia coli polymerase I can use 04 -methyldeoxythymidine or 5I—: methyldeoxythymidine in place of deoxythymidine in primed poly-(dA-dT)-poly-(dA-dT) synthesis. Proc.Natl.Acad.Sci. USA, 80: 4884–4888 (1983).CrossRefGoogle Scholar
  11. 11.
    Ding, R., Ghosh, K., Eastman, A. and Bresnick, E. DNA-mediated transfer and expression of a human DNA repair gene that demethylates 06 -methylguanine, Mol. Cell. Biol., 5: 3293–3296 (1985).Google Scholar
  12. 12.
    Goth-Goldstein, R. Inability of Chinese hamster ovary cells to excise 06 -alkylguanine, Cancer Res., 40: 2623–2624 (1980).PubMedGoogle Scholar
  13. 13.
    Pegg, A.E., Scicchitano D., and M.E. Dolan, Comparison of the rates of repair of 06 -alkylguanines in DNA by rat liver and bacterial 06 -alkylguanine-DNA alkyltransferase, Cancer Res. 44: 3806–3811 (1984).PubMedGoogle Scholar
  14. 14.
    Schimke, R.T., Gene amplification, drug resistance and cancer, Cancer Res., 44: 1735–1742 (1984).PubMedGoogle Scholar
  15. 15.
    Beranek, D.T., Heflich, R.H., Kodell, R.L., Morris, S.M. and Casciano, D.A. Correlation between specific DNA- methylation products and mutation induction at the HGPRT locus in Chinese hamster ovary cells, Mutation Res., 110: 171–180 (1983).PubMedCrossRefGoogle Scholar
  16. 16.
    Newbold, R.F., Warren, W., Medcalf, A.S.C. and Amos, J. Mutagenicity of carcinogenic methylating agents is associated with a specific DNA modification, Nature, 283 596–597 (1980).PubMedCrossRefGoogle Scholar
  17. 17.
    Natarajan, A.T., Simons, J.W.I.M., Vogel, E.W. and van Zeeland, A.A. Relationship between cell killing, chromosomal aberrations, sister-chromatid exchanges and point mutations induced by monofunctional alkylating agents in Chinese hamster cells. A correlation with different ethylation products, Mutation Res., 128: 31–40. (1984)PubMedCrossRefGoogle Scholar
  18. 18.
    Bignami, M., A. Vitelli, A. Di Muccio, M. Terlizzese, A. Calcagnile, G.A. Zapponi, P.H.M. Lohman, L. den Engelse and E. Dogliotti, Relationship between specific alkylated bases and mutations at two gene loci induced by ethylnitrosourea and diethyl sulfate in CHO cells. Mutat. Res. (in press).Google Scholar
  19. 19.
    Richardson, K.K., F.C. Richardson, F.C., R.M. Crosby, J.A. Swenberg and T.R. Skopek, DNA base changes and alkylation following in vivo exposure of Escherichia coli to N-methyl-N-nitrosourea or N-ethyl-N-nitrosourea Proc. Natl. Acad. Sci. 84: 344–348 (1987).CrossRefGoogle Scholar
  20. 20.
    Yarosh, D.B., Fornace, A.J., Day, R.S.III. Human 06 -alkylguanine DNA alkyltransferase fails to repair 04–methylthymine and methylphosphotriesters in DNA as efficiently as does the alkyltransferase from E. coli. Carcinogenesis, 6: 949–953 (1985).CrossRefGoogle Scholar
  21. 21.
    Domoradzki, J., Pegg, E. A., Dolan, M.E., Maher, V., and McCormick, J.J. Depletion of 06 -methylguanine-DNA- methyltransferase in human fibroblasts increases the mutagenic response to N-methyl-N-’nitro-N-nitrosoguanidine, Carcinogenesis 6: 1823–1826 (1985).PubMedCrossRefGoogle Scholar
  22. 22.
    Karran, P. and Williams, S.A. The cytotoxic and mutagenic effects of alkylating agents on human lymphoid cells are caused by different DNA lesions, Carcinogenesis 6: 789–792 (1985).PubMedCrossRefGoogle Scholar
  23. 23.
    Brennand, J. and Margison, G.P. Reduction of toxicity and mutagenicity of alkylating agents in mammalian cells harboring the Escherichia coli alkyltransferase gene, Proc. Natl. Acad. Sci. USA, 83: 6292–6296 (1986).PubMedCrossRefGoogle Scholar
  24. 24.
    Kataoka, H. Hall, J. and Karran, P. Complementation analysis of sensitivity to alkylating agents in Escherichia coli and Chinese hamster ovary cells by expression of a cloned bacterial DNA repair gene, EMBO J., 5: 3195–3200 (1986).Google Scholar
  25. 25.
    Brennand, J. and Margison, G.P. Expression in mammalian cells of a truncated Escherichia coli gene coding for 06–alkylguanine alkyltransferase reduces the toxic effects of alkylating agents, Carcinogenesis, 7: 2081–2084, (1986).PubMedCrossRefGoogle Scholar
  26. 26.
    Samson, L., Derfler, B. and Waldstein, E.A. Suppression of human DNA alkylation-repair defects by Escherichia coli DNA-repair genes, Proc. Natl. Acad. Sci. USA, 83: 5607–5610 (1986).PubMedCrossRefGoogle Scholar
  27. 27.
    Ishizaki, K., Tsujimura, T., Yawata, H., Fujio, C., Nakabeppu, Y., Sekiguchi, M. and Ikenaga, M. Transfer of E. coli 06 -methylguanine methyltransferase gene into repairdeficent human cells and restoration of cellular resistance to N-methyl-N’-nitro-N-nitrosoguanidine, Mutation. Res., 166: 139–141 (1986).Google Scholar
  28. 28.
    Goth-Goldstein, R., and M. Hughes, Characterization of a CHO variant in respect to alkylating agent-induced biological effects and DNA repair Mutat. Res. 184: 139–146 (1987).Google Scholar
  29. 29.
    Robson, C. and Hickson, I.D. Isolation of alkylating agent-sensitive Chinese hamster ovary cell lines Carcinogenesis 8: 601–605 (1987).Google Scholar
  30. 30.
    Samson, L. and Linn, S., DNA alkylation repair and the induction of cell death and sister chromatid exchange in human cells, Carcinogenesis, 8, 227–230 (1987).PubMedCrossRefGoogle Scholar
  31. 31.
    Ishida, R. and T. Takahashi N-methyl-N’-nitro-N- nitrosoguanidine-resistant HeLa S3 cells still have little 04 -methylguanine-DNA-methyltransferase activity and are hypermutable by alkylating agents Carcinogenesis 8: 1109–1113 (1987).Google Scholar
  32. 32.
    Kaina, B., A.A. Van Zeeland, C. Backendorf, H.W. Thielmann and P. van de Putte Transfer of human genes conferring resistance to methylating mutagens, but not to W irradiation and cross-linking agents, into Chinese hamster ovary cells. Mol.Cell.Biol. 7: 2024–2030 (1987).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • M. Bignami
    • 1
  • G. Aquilina
    • 1
  • A. Zijno
    • 1
  • G. Frosina
    • 2
  • A. Abbondandolo
    • 2
  • E. Dogliotti
    • 1
  1. 1.Istituto Superiore di Sanita’RomaItaly
  2. 2.Istituto Nazionale per la ricerca sul CancroGenovaItaly

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