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Removal of cyclobutane pyrimidine dimers from a UV-irradiated shuttle vector introduced into human cells

Abstract

A shuttle vector (pZH-1) carrying theE. coli lacZ gene under control of the SV40 early promoter was irradiated with UV and introduced into repair-proficient or repair-deficient human cell lines. The expression of irradiatedlacZ compared to unirradiatedlacZ was greater in repair-proficient cells (HT-1080) than in repair-deficient cells (XP12RO-SV40) belonging to xeroderma pigmentosum complementation group A. To ascertain whether the expression oflacZ in the repair-proficient cells was correlated with the removal of cyclobutane pyrimidine dimers (CPDs), we purified DNA from the recipient cells and used the CPD-specific enzyme T4 endonuclease V to measure the frequency of CPDs remaining in the plasmid as a whole and in two restriction fragments derived from it. We found that removal of CPDs occurred in both fragments in the repair-proficient cells but not in the repair-deficient cells. Our results provide the first direct evidence for the removal of CPDs from UV irradiated plasmids introduced into human cells and support the notion that expression of the UV-damagedlacZ gene in repair-proficient human cells reflects the removal of transcription blocking lesions from the gene.

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Literature Cited

  1. 1.

    Protić-Sabljić, M., and Kraemer, K.H. (1985).Proc. Natl. Acad. Sci. U.S.A. 82:6622–6626.

  2. 2.

    Klocker, H., Schneider, R., Burtscher, H.J., Auer, B., Hirsch-Kauffmann, M., and Schweiger, M. (1985).Eur. J. Cell. Biol. 39:346–351.

  3. 3.

    Cleaver, J.E., Vuksanovic, L., Player, A.N., and Lutze, L. (1989).Mutat. Res. 220:161–168.

  4. 4.

    Satokata, I., Tanaka, K., Miura, N., Narita, M., Mimaki, T., Satoh, Y., Kondo, S., and Okada, Y. (1992).Mutat. Res. 273:193–202.

  5. 5.

    Spivak, G., Leadon, S.A., Vos, J.-M., Meade, S., Hanawalt, P.C., and Ganesan, A.K. (1988).Mutat. Res. 193:97–108.

  6. 6.

    Ganesan, A., Spivak, G., and Hanawalt, P. (1983). InManipulation and Expression of Genes in Mammalian Cells, (eds.) Nagley, P., Linnane, A.W., Peacock, W.J., and Pateman, J.A. (Academic Press, Sidney, Australia), pp. 45–54.

  7. 7.

    Royer-Pokora, B., Peterson, W.D.J., and Haseltine, W.A. (1984).Exp. Cell Res. 151:408–420.

  8. 8.

    de Weerd-Kastelein, E.A., Keijzer, W., and Bootsma, D. (1972).Nature 238:80–83.

  9. 9.

    Cleaver, J.E., Cortés, F., Lutze, L.H., Morgan, W.F., Player, A.N., and Mitchell, D.L. (1987).Mol. Cell Biol. 7:3353–3357.

  10. 10.

    Lommel, L., and Hanawalt, P.C. (1993).Mol. Cell Biol. 13:970–976.

  11. 11.

    Ganesan, A.K., and Spivak, G. (1988). InDNA Repair: A Laboratory Manual of Research Procedures, Vol. 3, (eds.) Friedberg, E.C., and Hanawalt, P.C. (Marcel Dekker, New York), pp. 295–308.

  12. 12.

    An, G., Hidaka, K., and Siminovitch, L. (1982).Mol. Cell Biol. 2:1628–1632.

  13. 13.

    Seawell, P.C., and Ganesan, A.K. (1981). InDNA Repair: A Laboratory Manual of Research Procedures, Vol. 1B, (eds.) Friebberg, E.C., and Hanawalt, P.C. (Marcel Dekker, New York), pp. 425–430.

  14. 14.

    Graham, R., and van der Eb, A. (1973).Virology 52:456–467.

  15. 15.

    Parker, B., and Stark, G.R. (1979).J. Virol. 31:360–369.

  16. 16.

    Lommel, L., and Hanawalt, P.C. (1991).Mutat. Res. 255:183–191.

  17. 17.

    Mellon, I., and Bhorjee, J.S. (1982).Exp. Cell Res. 137:141–154.

  18. 18.

    Greenberg, M.E., and Ziff, E.B. (1984).Nature 311:433–438.

  19. 19.

    Smith, C.A., Cooper, P.K., and Hanawalt, P.C. (1981). InDNA Repair: A Laboratory Manual of Research Procedures, Vol. 1B, (eds.) Friedberg, E.C., and Hanawalt, P.C. (Marcel Dekker, New York), pp. 289–305.

  20. 20.

    Bohr, V.A., Smith, C.A., Okumoto, D.S., and Hanawalt, P.C. (1985).Cell 40:359–369.

  21. 21.

    Bohr, V.A., and Okumoto, D.S. (1988). InDNA Repair: A Laboratory Manual of Research Procedures, Vol. 3, (eds.) Friedberg, E.C., and Hanawalt, P.C. (Marcel Dekker, New York), pp. 347–366.

  22. 22.

    Vos, J.-M.H. (1988). InDNA Repair: A Laboratory Manual of Research Procedures, Vol. 3, (eds.) Friedberg, E.C., and Hanawalt, P.C. (Marcel Dekker, New York), pp. 367–398.

  23. 23.

    Purrello, M., and Balazs, I. (1983).Anal. Biochem. 128:393–397.

  24. 24.

    Rigby, P.W.J., Dieckmann, M., Rhodes, C., and Berg, P. (1977).J. Mol. Biol. 113:237–251.

  25. 25.

    Feinberg, A.P., and Vogelstein, B. (1984).Anal. Biochem. 137:266–267.

  26. 26.

    Mellon, I., and Hanawalt, P.C. (1989).Nature 342:95–98.

  27. 27.

    Spivak, G., Leadon, S.A., Vos, J.-M., Ganesan, A.K., and Hanawalt, P.C. (1987). InLectures and Symposia, 14th International Cancer Congress, Vol. 4, (Karger, Basel/Akadémiai Kiadó, Budapest), pp. 107–118.

  28. 28.

    Protić-Sabljić, M., and Kraemer, K.H. (1986).Photochem. Photobiol. 43:509–513.

  29. 29.

    Protić-Sabljić, M., and Kraemer, K.H. (1986).Carcinogenesis 7:1765–1770.

  30. 30.

    Protić, M., Riolides, E., Levine, A.S., and Dixon, K. (1988).Somat. Cell Mol. Genet. 14:351–357.

  31. 31.

    Fornace, A.J., Jr. (1992).Annu. Rev. Genet. 26:507–526.

  32. 32.

    Herrlich, P., Ponta, H., and Rahmsdorf, H.J. (1992).Rev. Physiol. Biochem. Pharmacol. 119:187–223.

  33. 33.

    Kalnins, A., Otto, K., Rüther, U., and Müller-Hill, B. (1983).EMBO J. 2:593–597.

  34. 34.

    Selby, C.P., and Sancar, A. (1990).J. Biol. Chem. 265:21330–21336.

  35. 35.

    Selby, C.P., and Sancar, A. (1993).J. Bacteriol. 175:7509–7514.

  36. 36.

    Vrieling, H., Venema, J., van Rooyen, M-L., van Hoffen, A., Menichini, P., Zdzienicka, M.Z., Simons, J.W.I.M., Mullenders, L.H.F., and van Zeeland, A.A. (1991).Nucleic Acids Res. 19:2411–2415.

  37. 37.

    Mellon, I., Spivak, G., and Hanawalt, P.C. (1987).Cell 51:241–249.

  38. 38.

    Sancar, A., and Tang, M.-S. (1993).Photochem. Photobiol. 57:905–921.

  39. 39.

    Hanawalt, P.C., and Mellon, I. (1993).Curr. Biol. 3:67–69.

  40. 40.

    Smerdon, M.J., Bedoyan, J., and Thoma, F. (1990).Nucleic Acids Res. 18:2045–2051.

  41. 41.

    Smerdon, M., and Thoma, F. (1990).Cell 61:675–684.

  42. 42.

    Leadon, S.A., and Lawrence, D.A. (1992).J. Biol. Chem. 267:23175–23182.

  43. 43.

    Sweder, K., and Hanawalt, P.C. (1992).Proc. Natl. Acad. Sci. U.S.A. 89:10696–10700.

  44. 44.

    Sun, Y. and Moses, R.E. (1991).Somat. Cell Mol. Genet. 17:229–238.

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Ganesan, A.K., Hanawalt, P.C. Removal of cyclobutane pyrimidine dimers from a UV-irradiated shuttle vector introduced into human cells. Somat Cell Mol Genet 20, 233–242 (1994). https://doi.org/10.1007/BF02254762

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Keywords

  • Human Cell
  • Human Cell Line
  • Recipient Cell
  • Shuttle Vector
  • lacZ Gene