The Expression of Bacterial DNA Alkylation Repair Enzymes in Mer- Human Cells

  • Leona Samson
  • Patrick Carroll
  • Bruce Derfler
  • William Rebeck


Simple monofunctional alkylating agents such as methylmethane sulphonate (MMS) and N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) cause a variety of alkylated lesions in DNA, and these lesions can cause the induction of mutation and cell death in both human and bacterial cells. The repair of these DNA alkylation damages reduces cell killing and the induction of mutations and chromosome damage. In E. coli, the repair of 06-methylguanine (06MeG) and 04-methylth5anine (04MeT) specifically prevents these lesions from causing G:C to A:T and A:T to G:C transition mutations, because if left unrepaired these lesions mispair during replication (1–6). On the other hand, the repair of N3-methylpurines (N3MeA and N3MeG) and 02-methylpyrimidines (02MeC and 02MeT) in E. coli specifically prevents cell killing (7–9). The repair of DNA alkylation damage in mammalian cells is somewhat less well understood, and it is not yet clear which alkylated lesions cause mutation and which cause cell death. Like E. coli, mammalian cells can repair 06alkylG, N3alkylA, N3alkylG, 04alkylT, 02alkylT and 02alkylC lesions (10–21). However, because observations have been made with a variety of cell types the results have not always been consistent. For instance, some studies indicated that 04MeT is repaired by rat liver cells (13,20,21), while others did not (22–24). Not all cell lines are able to repair all the alkylated lesions; for instance, CHO and V79 cells (25,26) and some human tumor cell lines (27–29) are unable to repair 06MeG. The ability of these particular cell lines to repair alkylated pyrimidines has not yet been measured. Other human cells, however, have been shown to repair 06MeG (19,27–29), 04alkylT, 02alkylT, 02alkylC (11), N3alkylA, N7alkylG (11,14,15), N3alkylG (15,30) and N7alkylA (15).


Sister Chromatid Exchange Methyl Methane Sulfonate alkB Gene Alkylation Damage Sister Chromatid Exchange Induction 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    A. Loveless, Possible relevance of 0–6 alkylation of deoxyguanosine to the mutagenicity and carcinogenicity of nitrosamines and nitrosamides. Nature (London) 223, 206–207 (1969).CrossRefGoogle Scholar
  2. 2.
    P. F. Schendel and P. E. Robins, Repair of 06-methylguanine in adapted Escherichia coli. Proc. Natl. Acad. Sci. USA 75, 6017–6020 (1978).PubMedCrossRefGoogle Scholar
  3. 3.
    P. Karran, T. Lindahl, and B. Griffin, Adaptive response to alkylating agents involves alteration in situ of 06-methylguanine residues in DNA. Nature (London) 280, 76–77 (1979).CrossRefGoogle Scholar
  4. 4.
    L. A. Dodson, R. S. Foote, S. Mitra, and W. E. Masker, Mutagenesis of bacteriophage T7 in vitro by Incorporation of 06-methylguanine during DNA synthesis. Proc. Natl. Acad. Sci. USA 79, 7440–7444 (1982).PubMedCrossRefGoogle Scholar
  5. 5.
    E. L. Loechler, C. L. Green, and J. M. Essigman, In vivo mutagenesis by 06-methylguanine built into a unique site in a virai genome. Proc. Nati. Acad. Sci. USA 81, 6271–6275 (1984).Google Scholar
  6. 6.
    B. D. Preston, B. Singer, and L. A. Loeb, Mutagenic potential of 04-methylthymine vivo determined by an enzymatic approach to site-specific mutagenesis. Proc. Natl. Acad. Sci. USA 8501–8505 (1986).Google Scholar
  7. 7.
    P. Karran, T. Lindahl, I. Ofsteng, G. B. Evensen, and E. Seeberg, Escherichia coli mutants deficient in 3-methyladenine-DNA glycosylase. J. Mol. Biol. 140, 101–127 (1980).PubMedCrossRefGoogle Scholar
  8. 8.
    P. Karran, T. Hjelmgren, and T. Lindahl, Induction of a DNA glycosylase for N-methylated purines is part of the adaptive response to alkylating agents. Nature (London) 296, 770–773 (1982).CrossRefGoogle Scholar
  9. 9.
    G. Evensen and E. Seeberg, Adaptation to alkylation resistance involves the induction of a DNA glycosylase. Nature (London) 296, 773–775 (1982).CrossRefGoogle Scholar
  10. 10.
    J. Shackleton, W. Warren, and J. J. Roberts, The excision of N-methyl-N-nitrosourea-induced lesions from the DNA of Chinese hamster cells as measured by the loss of sites sensitive to an enzyme extract that excises 3-methyIpurInes but not 06-methylguanine. Eur. J. Biochem 97, 425–433 (1979).PubMedCrossRefGoogle Scholar
  11. 11.
    W. J. Bodell, B. Singer, G. H. Thomas, and J. E. Cleaver, Evidence for removal at different rates of 0-ethyl pyrimidines and ethyl-phosphotriesters in two human fibroblast cell lines. Nucleic Acids Res. 6, 2819–2829 (1979).PubMedCrossRefGoogle Scholar
  12. 12.
    T. P. Brent, Partial purification and characterization of a human 3-methyladenine-DNA glycosylase. Biochemistry 8, 911–916 (1979).CrossRefGoogle Scholar
  13. 13.
    B. Singer, S. Spengler, and W. J. Bodell, Tissue-dependent enzyme-mediated repair or removal of 0-ethyl pyrimidines and ethyl purines in carcinogen-treated rats. Biochemistry 18, 911–916 (1981).Google Scholar
  14. 14.
    R. Cathcart and D. A. Goldthwait, Enzymatic excision of 3-methyl-adenosine and 7-methylguanine by a rat liver nuclear fraction. Biochemistry 19, 273–280 (1981).CrossRefGoogle Scholar
  15. 15.
    B. Singer and T. P. Brent, 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. USA 78, 856–860 (1981).PubMedCrossRefGoogle Scholar
  16. 16.
    G. P. Marglson, and A. E. Pegg, Enzymatic release of 7-methylguanine from methylated DNA by rodent liver extracts. Proc. Natl. Acad. Sci. USA 78, 861–865 (1981).CrossRefGoogle Scholar
  17. 17.
    J. M. Bogden, A. Eastman, and E. Bresnik, A system in mouse liver for the repair of 06-methylguanine lesions in methylated DNA. Nucleic Acids Res. 9, 3089–3092 (1981).PubMedCrossRefGoogle Scholar
  18. 18.
    J. R. Mehta, D. B. Ludlum, A. Renard, and W. G. Verly, Repair of 06-ethylguanine in DNA by a chromatin fraction from rat liver: transfer of the ethyl group to an acceptor protein. Proc. Natl. Acad. Sci. USA 78, 6766–6770 (1981).PubMedCrossRefGoogle Scholar
  19. 19.
    A. E. Pegg, M. Roberfroid, C. vonBahr, R. S. Foote, S. Mitra, H. Bresil, A. Likkacher, and R. Montesano, Removal of 06-methylguanine from DNA by human liver fractions. Proc. Natl. Acad. Sci. USA 79, 5162–5165 (1982).PubMedCrossRefGoogle Scholar
  20. 20.
    R. A. Becker and R. Montesano, Repair of 04-methyldeoxythymidine residues in DNA by mammalian liver extracts. Carcinogenesis 6, 313–317 (1985).PubMedCrossRefGoogle Scholar
  21. 21.
    L. D. Engelse, G. J. Menkveld, R. J. D. Brij, and A. D. Tates, Formation and stability of alkylated pyrimidines and purines (including amidazole ring-opened 7-alkylguanine) and alkyl-phosphotriesters in liver DNA of adult rats treated with ethylnitrosourea or dimethyl-nitrosamine. Carcinogenesis 7, 393–403 (1986).CrossRefGoogle Scholar
  22. 22.
    M. E. Dolan, D. Sciccitano, B. Singer, and A. E. Pegg, Comparison of repair of methylated pyrimidines in poly(dT) by extracts from rat liver and Escherichia coli. Biochem. Biophys. Res. Commun. 123, 324–330 (1984).PubMedCrossRefGoogle Scholar
  23. 23.
    M. E. Dolan and A. E. Pegg, Extent of formation of 04-methylthymidine in calf thymus DNA methylated by N-methyl-N-nitrosourea and lack of repair of this product by rat liver 06-alkylguanine-DNA-alkyltransferase. Carcinogenesis 6, 1611–1614 (1985).PubMedCrossRefGoogle Scholar
  24. 24.
    N. Huh and M. F. Rajewsky, Enzymatic elimination of 06-ethylguanine and stability of 04-ethylthymlne in the DNA of malignant neural cell lines exposed to N-ethyl-N-nitrosourea in culture. Carcinogenesis 7, 435–439 (1986).PubMedCrossRefGoogle Scholar
  25. 25.
    W. Warren, A. R. Crathom, and K. V. Shooter, The stability of methylated purines and of methylphosphotriesters in the DNA of V79 cells after treatment with N-methyl-N-nitrosourea. Biochem. Biophys. Res. Commun. 563, 82–88 (1979).Google Scholar
  26. 26.
    R. Goth-Goldstein, Inability of Chinese hamster ovary cells to excise 06-alkylguanine. Cancer Res. 40, 2623–2624 (1980).PubMedGoogle Scholar
  27. 27.
    R. S. Day III, C. H. J. Ziolkowski, D. A. Scuidero, S. A. Meyer, A. S. Lubiniecki, A. J. Girardi, S. M. Galloway, and G. D. Bynum, Defective repair of alkylated DNA by human tumour and SV40-transformed human cell strains. Nature (London) 288, 724–727 (1980).CrossRefGoogle Scholar
  28. 28.
    R. S. Day III, C. H. J. Ziolkowski, D. A. Scuidero, S. A. Meyer, and M. R. Mattem, Human tumor cell strains defective in the repair of alkylatlon damage. Carcinogenesis 1, 21–31 (1980).PubMedCrossRefGoogle Scholar
  29. 29.
    R. Sklar and B. Strauss, Removal of 06-methylguanlne from DNA of normal and xeroderma pigmentosum-derived lymphoblastoid lines. Nature (London) 289, 417–420 (1981).CrossRefGoogle Scholar
  30. 30.
    L. Samson and S. Linn, DNA alkylatlon repair and the induction of cell death and sister chromatid exchange in human cells. Carcinogenesis 8, 227–230 (1987).PubMedCrossRefGoogle Scholar
  31. 31.
    L. Samson and J. L. Schwartz, Evidence for an adaptive DNA repair pathway in CHO and human skin fibroblast cell lines. Nature (London) 287, 861–863 (1980).CrossRefGoogle Scholar
  32. 32.
    I. Teo, B. Sedgewick, M. W. Kilpatrick, T. V. McCarthy, and T. Lindahl, The intracellular signal for induction of resistance to alkylating agents in E. coll. Cell 45, 315–324 (1986).Google Scholar
  33. 33.
    P. Robins and J. Calms, Quantitation of the adaptive response to alkylating agents. Nature (London) 280, 74–76 (1979).CrossRefGoogle Scholar
  34. 34.
    T. Lindahl, B. Demple, and P. Robins, Suicide inactivation of the E. coli 06-methylguanine-DNA methyltransferase. EMBO J. 1, 1359–1363 (1982).PubMedGoogle Scholar
  35. 35.
    B. Demple, A. Jacobsson, M. Olsson, P. Robins, and T. Lindahl, Repair of alkylated DNA in Escherichia coll. J. Biol. Chem. 257, 13776–13780 (1982).PubMedGoogle Scholar
  36. 36.
    H. Kataoka, Y. Yamamoto, and M. Sekiguchi, A new gene (alkB) of scherichia coll that controls sensitivity to methyl methane sulfonate. J. Bacteriol. 153, 1301–1307 (1983).PubMedGoogle Scholar
  37. 37.
    Y. Nakabeppu, H. Kondo, and M. Sekiguchi, Cloning and characterization of the alkA gene of Escherichia coli that encodes 3-methyl-adenine DNA glycosylase II. J. Biol. Chem. 259, 13723–13729 (1984).PubMedGoogle Scholar
  38. 38.
    R. M. Baker, W. C. VanVoorhis, and L. A. Spencer, HeLa cell variants that differ in sensitivity to monofunctional alkylating agents, with independence of cytotoxic and mutagenic responses. Proc. Natl. Acad. Sci. USA 76, 5249–5253 (1979).PubMedCrossRefGoogle Scholar
  39. 39.
    D. B. Yarosh, A. J. Fomace, and R. S. Day, 06alkylguanlne-DNA alkyltransferase fails to repair 04methylthymine and methylphosphotriesters in DNA as efficiently as does the alkyltransferase from Escherichia coli. Carcinogenesis 6, 949–953 (1985).PubMedCrossRefGoogle Scholar
  40. 40.
    P. Karran, Possible depletion of a DNA repair enzyme in human lymphoma cells by subversive repair. Proc. Natl. Acad. Sci. USA 82, 5285–5289 (1985).PubMedCrossRefGoogle Scholar
  41. 41.
    P. Karran and S. A. Williams, 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
  42. 42.
    J. Domoradzki, A. E. Pegg, M. E. Dolan, V. M. Maher, and J. J. McCormick, 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
  43. 43.
    M. E. Dolan, C. D. Corsico, and A. E. Pegg, Exposure of HeLa cells to 06-alkylguanines increases sensitivity to the cytotoxic effects of alkylating agents. Biochem. Biophys. Res. Commun. 132, 178–185 (1985).PubMedCrossRefGoogle Scholar
  44. K. Ishizaki, T. Tsujimura, H. Yawata, C. Fujio, Y. Nakabeppu, M. Sekiguchi, and M. Ikenaga, Transfer of the coli 06-methyl-guanlne methyltransferase genes into repair deficient human cells and restoration of cellular resistance to N-methyl-N’-nitro-N-nitrosoguanidine. Mutat. Res. 166, 135–141 (1986).PubMedGoogle Scholar
  45. 45.
    H. Kataoka, J. Hall, and P. Karran, Complementation of sensitivity to alkylating agents in Escherichia coli and Chinese hamster ovary cells by expression a cloned bacterial DNA repair gene. EMBO J. 5, 3195–3200 (1986).PubMedGoogle Scholar
  46. 46.
    J. Brennand and G.P. Margison, Reduction of the 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

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Leona Samson
    • 1
  • Patrick Carroll
    • 1
  • Bruce Derfler
    • 1
  • William Rebeck
    • 1
  1. 1.Charles A. Dana Laboratory of ToxicologyHarvard School of Public HealthBostonUSA

Personalised recommendations