Detection of Platinum Lesions at the Nucleotide Level in Cells using Single Strand Ligation PCR

  • John A. Hartley
  • Robert L. Souhami
  • Keith A. Grimaldi


The mechanism of action of many anti-cancer drugs, including platinum based drugs, involves direct, covalent adduct formation on nucleotides in DNA. Cisplatin forms mainly intrastrand cross-links between two adjacent guanines in runs of two or more consecutive guanines and between AG pairs.1,2 Recently, the sequence preferences of lesion formation have been studied in isolated DNA3 by exploiting the finding that such lesions can block the progression of DNA taq polymerases. This and other assays using plasmid DNA have revealed that even relatively simple DNA damaging agents such as cisplatin show a degree of sequence preference in adduct formation.4,5 Studying drug interactions in artificial systems with oligonucleotides or with plasmid DNA provides useful information but cannot necessarily predict intracellular behaviour where DNA exists in a highly ordered structure, complexed with many proteins, and where other cellular components may affect reactivity.


Single Copy Gene Sequence Preference Intrastrand Crosslinks Consecutive Guanine Covalent Adduct Formation 


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  1. 1.
    Fichtinger-Schepman, A. M. J., Van Der Meer, J. L., Den Hartag, J. H. J., Loman, P. H. M. and Reedijk, J. Biochemistry, 24:707 (1985).CrossRefPubMedGoogle Scholar
  2. 2.
    Roberts, J. J. and Friedlos, F. Biochim. Biophys. Acta, 655:146 (1981).CrossRefPubMedGoogle Scholar
  3. 3.
    Ponti, M., Forrow, S. F., Souhami, R. L., D’Incalci, M. and Hartley, J. A. Nucleic Acids Res., 19:2929 (1991).CrossRefPubMedGoogle Scholar
  4. 4.
    Murray, V., Motyka, H., England, P. R., Wickham, G., Lees, H. H., Denny, W. A. and McFadyen, W. A. J. Biol Chem, 267: 18805 (1992).PubMedGoogle Scholar
  5. 5.
    Cullinane, C., Wickham, G., McFadyen, W. D., Denny, W. A., Palmer, B. D. and Phillips, D. R. Nucleic Acids Res., 21:393 (1993).CrossRefPubMedGoogle Scholar
  6. 6.
    Kohn, K. W., Hartley, J. A. and Mattes, W. B. Nucleic Acid Res, 15:10531 (1987).CrossRefPubMedGoogle Scholar
  7. 7.
    Fichtinger-Schepman, A-M J., van Oosterom, A.T., Lohman, P.H.M., and Berends, F.: Cancer Res., 47:3000 (1987).PubMedGoogle Scholar
  8. 8.
    Parker, R.J., Gill, I., Tarone, R., Vionnet, J.A., Grunberg, S., Muggia, F.M., and Reed, E.: Carcinogenesis, 12:1253 (1991).CrossRefPubMedGoogle Scholar
  9. 9.
    Bohr BA, Philips DH and Hanawalt PC, Cancer Res 47:6426 (1987).PubMedGoogle Scholar
  10. 10.
    Bohr BA Carcinogenesis 11:1983 (1991).CrossRefGoogle Scholar
  11. 11.
    Wassermann K, Ǩohn KW and Bohr VA J Biol Chem 265:13906 (1990).PubMedGoogle Scholar
  12. 12.
    Futscher BW, Pieper RO, Dalton WS and Erickson LC Cell Growth and Differentiation 3:217 (1992).PubMedGoogle Scholar
  13. 13.
    Larminat F, Zhen W and Boh VA J Biol Chem 268:2649 (1993).PubMedGoogle Scholar
  14. 14.
    Jennerwein, M. M. and Eastman, A. Nucleic Acids Res., 19:6209 (1991).CrossRefPubMedGoogle Scholar
  15. 15.
    Kalinowsky, D. P., Illenye, S. and Van Houten, B. Nucleic Acids Res., 20:3485 (1992).CrossRefGoogle Scholar
  16. 16.
    Grimaldi KA, Bingham JP, Souhami RL and Hartley JA Analytical Biochem. 222:236 (1994).CrossRefGoogle Scholar
  17. 17.
    Hartley, J. A., Bingham, J. P. and Souhami, R. L. Nucleic Acid Res., 20:3175 (1992).CrossRefPubMedGoogle Scholar
  18. 18.
    Murray, V., Motyka, H., England, P. R., Wickham, G., Lee, H. H., Denny, W. A. and McFadyen, W. D. Biochemistry, 31:11812 (1992).CrossRefPubMedGoogle Scholar
  19. 19.
    Bubley, G.J., Ogata, G.K., Dupuis, N.P., and Teicher, B.A. Cancer Res. 54:6325 (1994)PubMedGoogle Scholar
  20. 20.
    Pfeifer, G. P., Steigerwald, S. D., Mueller, P. R., Wold, B. and Riggs, A. D. Science, 246:810 (1989).CrossRefPubMedGoogle Scholar
  21. 21.
    Mueller, P. R. and Wold, B. Science, 246:780 (1989).CrossRefPubMedGoogle Scholar
  22. 22.
    Tessier, D. C., Brousseau, R. and Vernet, T. Analyt. Biochem, 158:171 (1986).CrossRefPubMedGoogle Scholar
  23. 23.
    Grimaldi, K.A., McAdam, S.R., Souhami, R.L., and Hartley, J.A. Nucleic Acids Res., 22:2311 (1994).CrossRefPubMedGoogle Scholar
  24. 24.
    Grimaldi, K.A., McAdam, S.R. and Hartley, J.A.: In: Technologies for Detection of DNA Damage and Mutations., Pfeifer, G.P. (ed), Plenum Press, New York, in press, 1996.Google Scholar
  25. 25.
    Smellie, M., Grimaldi, K.A., Bingham, J.P., McAdam, S.R., Thompson, A.S., Thurston, D.E., and Hartley, J.A. Proc.Am. Assoc. Cancer Res., 35, 536 (1994)Google Scholar
  26. 26.
    Comess, K. M., Burstyn, J. N., Essigman, J. M. and Lippard, S. L. Biochemistry, 31:3975 (1992).CrossRefPubMedGoogle Scholar
  27. 27.
    Fichtinger-Schepman, A. M. J., Van Oosterom, A. T. and Lohman, P. H. M. Cancer Res, 47:3000 (1987).PubMedGoogle Scholar
  28. 28.
    Bernges, F. and Holler, E. Nucleic Acid Res, 19:1483 (1991).CrossRefPubMedGoogle Scholar
  29. 29.
    Marrot, L. and Leng, M. Biochemistry, 28:1454 (1989).CrossRefPubMedGoogle Scholar
  30. 30.
    Sherman, S. E. and Lippard, S. J. Chem. Rev, 81:1153 (1987).CrossRefGoogle Scholar
  31. 31.
    Shwartz, H., Shavitt, O. and Livneh, Z. J. Biol. Chem, 263:18277 (1988).PubMedGoogle Scholar
  32. 32.
    Lindahl, T. Nature, 362:709 (1993).CrossRefPubMedGoogle Scholar
  33. 33.
    Mellon, I., Spivak, G. and Hanawalt, P. C. Cell, 51:241 (1987).CrossRefPubMedGoogle Scholar
  34. 34.
    Selby, C. P. and Sancar, A. Science, 260:53 (1993).CrossRefPubMedGoogle Scholar
  35. 35.
    Bohr, V. A., Smith, C. A., Okumoto, D. S. and Hanawalt, P. C. Cell 40:359 (1985).CrossRefPubMedGoogle Scholar
  36. 36.
    Hanawalt, P. C. Genome, 31:605 (1989).CrossRefPubMedGoogle Scholar
  37. 37.
    Bohr, V. Carcinogenesis, 12:1983 (1991).CrossRefPubMedGoogle Scholar
  38. 38.
    Mellon, I. and Hanawalt, P. C. Nature, 342:95 (1989).CrossRefPubMedGoogle Scholar
  39. 39.
    Lommel, L. and Hanawalt, P. C. Mol Cell Biol, 13:970 (1993).PubMedGoogle Scholar
  40. 40.
    Kunala, S. and Brash, D. E. Proc. Natl Acad. Sci. U.S.A., 89:11031 (1992).CrossRefPubMedGoogle Scholar
  41. 41.
    Tornaletti S and Pfeifer GP Science 263:1436 (1994).CrossRefPubMedGoogle Scholar
  42. 42.
    Gao S, Drovin R and Holmquist Science 263:1438 (1994).CrossRefPubMedGoogle Scholar
  43. 43.
    Zhen, W., Link, C. J., O’Connor, P. M., Reed, E., Parker, R., Howell, S. B. and Bohr, V. A. Molec. Cell. Biol., 12:3689 (1992).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • John A. Hartley
    • 1
  • Robert L. Souhami
    • 1
  • Keith A. Grimaldi
    • 1
  1. 1.Department of OncologyUniversity College London Medical SchoolLondonUK

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