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The Use of DNA-Repair-Deficient Mutants of Chinese Hamster Ovary Cells in Studying Mutagenesis Mechanisms and Testing for Environmental Mutagens

  • Larry H. Thompson
Part of the Basic Life Sciences book series (volume 15)

Summary

Our laboratory has taken a somatic-cell-genetics approach to the study of mutagenesis by utilizing mutant strains of Chinese hamster ovary (CHO) cells that are deficient in DNA repair processes. From more than 150 UV-sensitive strains tested, five complementation classes were identified, and representative mutants were found to be defective at, or before, the incision step of excision repair. A representative mutant, strain UV-5, was compared with the parental strain in terms of cytotoxicity and dose-response curves for mutation induction after treatment with UV and several chemicals that are known to produce large adducts in DNA. Excision repair in normal CHO cells protects against both cytotoxicity and mutagenesis, but the degree of protection depends on both the agent and the genetic marker used for detecting mutations. Upon treatment with low doses (100% cell survival) of the polyaromatic hydrocarbon 7-romomethylbenz(a)anthracene, repair-deficient UV-5 cells had linear responses for mutation induction to thioguanine resistance or azaadenine resistance, whereas the normal repair-proficient cells showed curvilinear responses in which the slope increased with dose. This behavior suggests that in the normal cells the repair system acting on potentially mutagenic lesions becomes saturated at doses that produce cytotoxicity. In no instance was a lower mutation frequency induced in UV-5 cells than the parental cells, at a given dose of mutagen, suggesting that the excision repair system is error-free in normal CHO cells.

Keywords

Chinese Hamster Ovary Cell Chinese Hamster Ovary Excision Repair Sister Chromatid Exchange Mutation 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.

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References

  1. 1.
    Hollstein, M. and McCann J.: Short-term tests for carcinogens and mutagens. Mutat. Res., 65: 133–226 (1979).PubMedGoogle Scholar
  2. 2.
    Ames, B.N.: Identifying environmental chemicals causing mutations and cancer. Science, 204: 587–593 (1979).PubMedCrossRefGoogle Scholar
  3. 3.
    Hsie, A.W.: O’Neill, J.P., Couch, D.B., SanSebastian, J.R., Brimer, P.A., Machanoff, R., Fuscoe, J.C., Riddle, J.C., Li, A.P., Forbes, N.L., and Hsie, M.H.: Quantitative analysis of radiation-and chemical-induced lethality and mutagenesis in Chinese hamster ovary cells. Radiat. Res., 76: 471–492 (1978).Google Scholar
  4. 4.
    Clive, D., Johnson, K.O., Spector, J.F.S., Batson, A.G., and Brown, M.M.M.: Validation and characterization of the L5178Y/TK+/- mouse lymphoma mutagen assay system. Mutat. Res., 59: 61–108 (1979).PubMedCrossRefGoogle Scholar
  5. 5.
    Hsie, A.W., O’Neill, J.P., and McElheny, V.K., Eds.: Banbury Report 2, Mammalian Cell Mutagenesis: The Maturation of Test Systems. Cold Spring Harbor Laboratory (1979).Google Scholar
  6. 6.
    Wolff, S. and Perry, P.: Differential Giemsa staining of sister chromatids and the study of sister chromatid exchanges without autoradiography. Chromosoma, 48: 341–353 (1974).PubMedCrossRefGoogle Scholar
  7. 7.
    Perry, P.E.: Chemical mutagens and sister chromatid exchange. Vol. 6 in Chemical Mutagens, Ed. by F.J. deSerres and A. Hollaender. Plenum Press, New York, pp. 1–39 (1980).Google Scholar
  8. 8.
    Carrano, A.V., Minkler, J.L., Stetka, D.G., and Moore II, D.H.: Variation in the baseline sister chromatid exchange frequency in human lymphocytes. Environmental Mutagen., 2: 325–337 (1980).CrossRefGoogle Scholar
  9. 9.
    Carrano, A.V. and Moore II, D.H.: The rationale and methodology for quantifying siter chromatid exchange in humans. In Mutagenicity: From Bacteria to Man. Ed. by J.A. Meddle. Academic Press, New York, in press.Google Scholar
  10. 10.
    Strauss, G.H. and Albertini, R.J.: Enumeration of 6-thiog anine-resistant peripheral blood lymphocytes in man as a potential test for somatic cell mutations arising in vivo. Mutat. Res., 61: 353–379 (1979).PubMedCrossRefGoogle Scholar
  11. 11.
    Ansari, A.A., Baig, M.A., and Malling, H.V.: In Vivo germinal mutation detection with “monospecific” antibody against lactate dehydrogenase-X. Proc. Natl. Acad. Sci. USA, 77: 7352–7356 (1980).PubMedCrossRefGoogle Scholar
  12. 12.
    Ansari, A.A., Baig, M.A., and Malling, H.V.: Development of in vivo somatic mutation system using antibody against hemoglobin. Preparation and use of an anti-hemoglobin antibody for identifying C57BL/6 red cells in artificial mixture of DBA/2 and C57BL/6 red cells, Mutat. Res., 81: 243–255 (1981).Google Scholar
  13. 13.
    Yang, L.L., Maher, V.M., and McCormick J.J.: Error-free excision of the cytotoxic, mutagenic N -deoxyguanosine DNA adduct formed in human fibroblasts by (±)-7ß, 8adihydroxy-9a, 10a-epoxy-7,8,9,10-tetrahydro-benzo(a)pyrene. Proc. Natl. Acad. Sci. USA, 77: 5933–5937 (1980).Google Scholar
  14. 14.
    Maher, V.M., Dorney, D.J., Mendrala, A.L., Konze-Thomas, B., and McCormick, J.J.: DNA excision-repair processes in human cells can eliminate the cytotoxic and mutagenic consequences of ultraviolet irradiation. Mutat. Res., 62: 311–323 (1979).PubMedCrossRefGoogle Scholar
  15. 15.
    Cleaver, J.E.: Xeroderma pigmentosum. In Metabolic Basis of Inherited Disease, Ed. by J.B. Stanbury, J.B. Wyngaarden, and D.S. Fredrickson, 4th edition, McGraw-Hill, New York, pp. 1072–1095 (1978).Google Scholar
  16. 16.
    Maher, V.M., McCormick, J.J., Grover, P.L., and Sims, P.: Effect of DNA repair on the cytotoxicity and mutagenicity of polycyclic hydrocarbon derivatives in normal and xeroderma pigmentosum human fibroblasts. Mutat. Res., 43: 117–138 (1977).PubMedCrossRefGoogle Scholar
  17. 17.
    Setlow, R.B.: Repair deficient human disorders and cancer. Nature, 271: 713–717 (1978).PubMedCrossRefGoogle Scholar
  18. 18.
    Simons, J.W.I.M.: Development of a liquid-holding technique for the study of DNA-repair in human diploid fibroblasts. Mutat. Res., 59: 273–283 (1979).PubMedCrossRefGoogle Scholar
  19. 19.
    Sarasin, A. and Benoit, A.: Induction of an error-prone mode of DNA repair in UV-irradiated monkey kidney cells. Mutat. Res., 70: 71–81 (1980).PubMedCrossRefGoogle Scholar
  20. 20.
    DasGupta, U.B. and Summers, W.C.: Ultraviolet reactivation of herpes simplex virus is mutagenic and inducible in mammalian cells. Proc. Natl. Acad. Sci. USA, 75: 2378–2381 (1978).PubMedCrossRefGoogle Scholar
  21. 21.
    Laval, F.: Effect of uncouplers on radiosensitivity and mutagenicity in x-irradiated mammalian cells, Proc. Natl. Acad. Sci. USA, 77: 2702–2725 (1980).PubMedCrossRefGoogle Scholar
  22. 22.
    Corsaro, C.M. and Migeon, B.R.: Comparison of contact-mediated communication in normal and transformed human cells in culture. Proc. Natl. Acad. Sci. USA, 74: 4476–4480 (1977).PubMedCrossRefGoogle Scholar
  23. 23.
    Siminovitch, L.: On the nature of heritable variation in cultured somatic cells. Cell, 7: 1–11 (1976).PubMedCrossRefGoogle Scholar
  24. 24.
    Gautschi, J.R., Young, B.R., and Cleaver, J.E.: Repair of damaged DNA in the absence of protein synthesis in mammalian cells. Exptl. Cell Res., 76: 87–94 (1973).PubMedCrossRefGoogle Scholar
  25. 25.
    Meyn, R.E., Vizard, D.L., Hewitt, R.R., and Humphrey, R.M.: The fate of pyrimidine dimers in the DNA of ultraviolet-irradiated Chinese hamster cells. Photochem. and Photobiol., 20: 221–226 (1974).CrossRefGoogle Scholar
  26. 26.
    Dipple, A. and Roberts, J.J.: Excision of 7-bromomethylbenz(a)anthracene-DNA adducts in replicating mammalian cells. Biochem., 16: 1499–1503 (1977).CrossRefGoogle Scholar
  27. 27.
    Thompson, L.H., Rubin, J.S., Cleaver, J.E., Whitmore, G.F., and Brookman, K.: A screening method for isolating DNA repair-deficient mutants of CHO cells. Somat. Cell Genet., 6: 391–405 (1980).PubMedCrossRefGoogle Scholar
  28. 28.
    Busch, D.B., Cleaver, J.E., and Glaser, D.A.: Large-scale isolation of UV-sensitive clones of CHO cells. Somat. Cell Genet., 6: 407–418 (1980).PubMedCrossRefGoogle Scholar
  29. 29.
    Thompson, L.H., Busch, D.B., Brookman, K., Mooney, C.L., and Glaser, D.A.: Genetic diversity of ultraviolet-sensitive DNA repair mutants of Chinese hamster ovary cells. Proc. Natl. Acad. Sci. USA, in press (1981).Google Scholar
  30. 30.
    Brookman, K.W., Thompson, L.H., Salazar, E.P., Dillehay, L.E., and Mooney, C.L.: Genetic complementation, DNA repair, and mutation induction in UV-sensitive Chinese hamster ovary cell mutants. 12th Ann. Mtg. Environ. Mutagen Soc., p. 161, abstract (1981).Google Scholar
  31. 31.
    Keijzer, W., Jaspers, N.G.J., Abrahams, P.J., Taylor, A.M.R., Arlett, C.F., Zelle, B., Takebe, H., Kinmont, P.D.S., and Bootsma; D.: A seventh complementation group in excision-deficient xeroderma pigmentosum. Mutat. Res., 62: 183–190 (1979).Google Scholar
  32. 32.
    Arase, S., Kozuka, T., Tanaka, K., Ikenaga, M., and Takebe, H.: A sixth complementation group in xeroderma pigmentosum. Mutat. Res., 59: 143–146 (1979).PubMedCrossRefGoogle Scholar
  33. 33.
    Arlett, C.F. and Lehman, A.R.: Human disorders showing increased sensitivity to the induction of genetic damage. Ann. Rev. Genet., 12: 95–115 (1978).PubMedCrossRefGoogle Scholar
  34. 34.
    Thompson, L.H., Brookman, K.W., Dillehay, L.E., Carrano, A.V., Mooney, C.L., Mazrimas, J.A., and Minkler, J.A.: A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister chromatid exchange. Mutat. Res., submitted.Google Scholar
  35. 35.
    Thompson, L.H., Fong, S., and Brookman, K.: Validation of conditions for efficient detection of HPRT and APRT mutations in suspension-cultured Chinese hamster ovary cells. Mutat. Res., 74: 21–36 (1980).PubMedGoogle Scholar
  36. 36.
    Krahn, D.F. and Heidelberger, C.: Liver homogenate-mediated mutagenesis in Chinese hamster V79 cells by polycyclic aromatic hydrocarbons and aflatoxins. Mutat. Res., 46: 27–44 (1977).PubMedGoogle Scholar
  37. 37.
    Venitt, S. and Tarmy, E.M.: The selective excision of arylalkylated products from the DNA of Escherichia coli treated with the carcinogen 7-bromomethylbenz(a)anthracene. Biochim. Biophys. Acta, 287: 38–51 (1972).PubMedGoogle Scholar
  38. 38.
    Thompson, L.H., Brookman, K.W., and Carrano, A.V.: The role of DNA repair in mutagenesis of Chinese hamster ovary cells by 7-bromomethylbenz(a)anthracene. Proc. Natl. Acad. Sci. USA, submitted.Google Scholar
  39. 39.
    Kohn, K.W., Erickson, L.C., Ewig, R.A.G., and Freidman, C.A.: Fractionation of DNA from mammalian cells by alkaline elution. Biochem., 15: 4629–4637 (1976).CrossRefGoogle Scholar
  40. 40.
    Hiss, E.A. and Preston, R.J.: The effect of cytosine arabinoside on the frequency of single-strand breaks in DNA of mammalian cells following irradiation or chemical treatment. Biochim. Biophys. Acta., 478: 1–8 (1977).PubMedGoogle Scholar
  41. 41.
    Fornace, A.J., Kohn, K.W., and Kann Jr., H.E.: DNA single-strand breaks during repair of UV damage in human fibroblasts and abnormalities of repair in xeroderma pigmentosum. Proc. Natl. Acad. Sci. USA, 73: 39–43 (1976).PubMedCrossRefGoogle Scholar
  42. 42.
    Tanaka, K., Hayakawa, H., Sekiguchi, M., and Okada, Y.: Specific action of T4 endonuclease V on damaged DNA in xeroderma pigmentosum cells in vivo. Proc. Natl. Acad. Sci. USA, 74: 2958–2962 (1977).PubMedCrossRefGoogle Scholar
  43. 43.
    Bootsma, D.: Xeroderma pigmentosum. In DNA Repair Mechanisms, Ed. by P.C. Hanawalt, E.C. Friedberg, and C.F. Fox. (Proc. ICN-UCLA Symp. on DNA Repair Mechanisms, February 1978, Keystone, Colo.) Academic Press, New York, p. 589–601 (1978).Google Scholar
  44. 44.
    Riddle, J.C. and Hsie, A.W.: An effect of cell-cycle position on ultraviolet-light-induced mutagenesis in Chinese hamster ovary cells. Mutat. Res., 52:’409–420 (1978).Google Scholar
  45. 45.
    McCormick, J.J. and Maher, V.M.: Mammalian cell mutagenesis as a biological consequence of DNA damage. In DNA Repair Mechanisms, Ed. by P.C. Hanawalt, E.C. Friedberg, and C.F. Fox. (Proc. ICN-UCLA Symp. on DNA Repair Mechanisms, February 1978, Keystone, Colo.) Academic Press, p. 739–749 (1978).Google Scholar
  46. 46.
    Munson, R.J., and Goodhead, D.T.: The relation between induced mutation frequency and cell survival0000000a theoretical approach and an examination of experimental data for eukaryotes. Mutat. Res., 42: 145–160 (1977).PubMedCrossRefGoogle Scholar
  47. 47.
    McCaw, B.A., Dipple, A., Young, S., and Roberts, J.J.: Excision of hydrocarbon-DNA adducts and consequent survival in normal and repair defective human cells. Chem.-Biol. Interactions, 22: 139–151 (1978).Google Scholar
  48. 48.
    Dipple, A., Brookes, P., Mackintosh, D.S., and Rayman, M.P.: Reaction of 7-bromomethylbenz(a)anthracene with nucleic acids, polynucleotides, and nucleosides. Biochem., 10: 4323–4330 (1971).CrossRefGoogle Scholar
  49. 49.
    Sugimura, T., Kawachi, T., Nagao, M., Yahagi, T., Seino, Y., Okamoto, T., Shudo, K., Kosuge, T., Tsuji, K., Wakabayshi, K., Iitaka, Y., and Itai, A.: Mutagenic principle(s) in tryptophan and phenylalanine pyrolysis products. Proc. Japan Acad., 53: 58–61 (1977).CrossRefGoogle Scholar
  50. 50.
    Sugimura, T.: Naturally occurring genotoxic carcinogens. In Naturally Occurring Carcinogens-Mutagens and Modulators of Carcinoßenesis, Ed. by E.C. Miller et al., Japan Sci. Press, Tokyo/Univ. Park Press, Baltimore, pp. 241–261 (1979).Google Scholar
  51. 51.
    Meyn, R.E., Jenkins, S.L., and Thompson, L.H.: Defective removal of DNA-cross-links in a repair-deficient mutant of Chinese hamster cells. Cancer Res., submitted for publication.Google Scholar
  52. 52.
    Fujiwara, Y., Tatsumi, M., and Sasaki, M.S.: Cross-link repair in human cells and its possible defect in Fanconi’s anemia cells. J. Mol. Biol., 113: 635–649 (1977).PubMedCrossRefGoogle Scholar
  53. 53.
    Kaye, J. Smith, C.A., and Hanawalt, P.C.: DNA repair in human cells containing photoadducts of 8-methoxypsoralen or angelicin. Cancer Res., 40: 696–702 (1980).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1983

Authors and Affiliations

  • Larry H. Thompson
    • 1
  1. 1.Biomedical Sciences DivisionL-452 Lawrence Livermore National LaboratoryLivermoreUSA

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