Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1994 Jul 11;22(13):2519–2524. doi: 10.1093/nar/22.13.2519

Mutagenesis in monkey cells of a vector containing a single d(GPG) cis-diamminedichloroplatinum(II) adduct placed on codon 13 of the human H-ras proto-oncogene.

M J Pillaire 1, A Margot 1, G Villani 1, A Sarasin 1, M Defais 1, A Gentil 1
PMCID: PMC308204  PMID: 8041613

Abstract

Cisplatin (cis-[Pt(NH3)2Cl2]) is a widely used antitumor agent whose mutagenic activity raises the possibility of the induction of secondary cancer as a result of treatment. Mutation of the proto-oncogene H-ras is found in more than 30% of all human tumors, where it has been postulated to contribute to the initiation and progression of human cancers. Activating mutations in the H-ras gene are predominantly single-base substitutions, most frequently at codons 12, 13 and 61. In the present work we have studied the mutational spectra induced by a single cis-[Pt(NH3)2d(GpG)] adduct, the most frequent DNA crosslink formed by cisplatin. We have constructed a 25-mer-Pt oligonucleotide singly modified at codon 13 (GGT) within the human H-ras DNA sequence and we have inserted it into a single-stranded SV40-based shuttle vector able to replicate in simian COS7 cells. After replication in the mammalian host, vectors were extracted, amplified in bacteria and DNA from 124 randomly chosen colonies was sequenced. The observed mutation frequency was 21%. Base substitutions were the most frequent modification. 92% of the mutagenic events occurred at one or both of the platinated guanines of codon 13. The single G-->T transversion accounted for 65% of the total mutations scored. All single base substitutions were located at the G in the 3' position showing, for the first time, that the guanine at the 3' side of a cis-[Pt(NH3)2d(GpG)] adduct may be a preferential site for cisplatin induced mutations. The substitution G-->T at this position of the codon 13 of the H-ras proto-oncogene is known to induce the oncogenic properties of the p21ras protein.

Full text

PDF
2519

Images in this article

2522Image
on p.2522

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  2. Basu A. K., Essigmann J. M. Site-specifically modified oligodeoxynucleotides as probes for the structural and biological effects of DNA-damaging agents. Chem Res Toxicol. 1988 Jan-Feb;1(1):1–18. doi: 10.1021/tx00001a001. [DOI] [PubMed] [Google Scholar]
  3. Belguise-Valladier P., Maki H., Sekiguchi M., Fuchs R. P. Effect of single DNA lesions on in vitro replication with DNA polymerase III holoenzyme. Comparison with other polymerases. J Mol Biol. 1994 Feb 11;236(1):151–164. doi: 10.1006/jmbi.1994.1125. [DOI] [PubMed] [Google Scholar]
  4. Bos J. L. ras oncogenes in human cancer: a review. Cancer Res. 1989 Sep 1;49(17):4682–4689. [PubMed] [Google Scholar]
  5. Brabec V., Balcarova Z. Restriction-enzyme cleavage of DNA modified by platinum(II) complexes. Eur J Biochem. 1993 Aug 15;216(1):183–187. doi: 10.1111/j.1432-1033.1993.tb18131.x. [DOI] [PubMed] [Google Scholar]
  6. Bradley L. J., Yarema K. J., Lippard S. J., Essigmann J. M. Mutagenicity and genotoxicity of the major DNA adduct of the antitumor drug cis-diamminedichloroplatinum(II). Biochemistry. 1993 Jan 26;32(3):982–988. doi: 10.1021/bi00054a031. [DOI] [PubMed] [Google Scholar]
  7. Brouwer J., van de Putte P., Fichtinger-Schepman A. M., Reedijk J. Base-pair substitution hotspots in GAG and GCG nucleotide sequences in Escherichia coli K-12 induced by cis-diamminedichloroplatinum (II). Proc Natl Acad Sci U S A. 1981 Nov;78(11):7010–7014. doi: 10.1073/pnas.78.11.7010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bubley G. J., Ashburner B. P., Teicher B. A. Spectrum of cis-diamminedichloroplatinum(II)-induced mutations in a shuttle vector propagated in human cells. Mol Carcinog. 1991;4(5):397–406. doi: 10.1002/mc.2940040512. [DOI] [PubMed] [Google Scholar]
  9. Burnouf D., Duane M., Fuchs R. P. Spectrum of cisplatin-induced mutations in Escherichia coli. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3758–3762. doi: 10.1073/pnas.84.11.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Burnouf D., Gauthier C., Chottard J. C., Fuchs R. P. Single d(ApG)/cis-diamminedichloroplatinum(II) adduct-induced mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6087–6091. doi: 10.1073/pnas.87.16.6087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Calsou P., Frit P., Salles B. Repair synthesis by human cell extracts in cisplatin-damaged DNA is preferentially determined by minor adducts. Nucleic Acids Res. 1992 Dec 11;20(23):6363–6368. doi: 10.1093/nar/20.23.6363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cariello N. F., Swenberg J. A., Skopek T. R. In vitro mutational specificity of cisplatin in the human hypoxanthine guanine phosphoribosyltransferase gene. Cancer Res. 1992 May 15;52(10):2866–2873. [PubMed] [Google Scholar]
  13. Chu G. Cellular responses to cisplatin. The roles of DNA-binding proteins and DNA repair. J Biol Chem. 1994 Jan 14;269(2):787–790. [PubMed] [Google Scholar]
  14. Comess K. M., Burstyn J. N., Essigmann J. M., Lippard S. J. Replication inhibition and translesion synthesis on templates containing site-specifically placed cis-diamminedichloroplatinum(II) DNA adducts. Biochemistry. 1992 Apr 28;31(16):3975–3990. doi: 10.1021/bi00131a013. [DOI] [PubMed] [Google Scholar]
  15. Corda Y., Job C., Anin M. F., Leng M., Job D. Transcription by eucaryotic and procaryotic RNA polymerases of DNA modified at a d(GG) or a d(AG) site by the antitumor drug cis-diamminedichloroplatinum(II). Biochemistry. 1991 Jan 8;30(1):222–230. doi: 10.1021/bi00215a032. [DOI] [PubMed] [Google Scholar]
  16. Diwan B. A., Anderson L. M., Rehm S., Rice J. M. Transplacental carcinogenicity of cisplatin: initiation of skin tumors and induction of other preneoplastic and neoplastic lesions in SENCAR mice. Cancer Res. 1993 Sep 1;53(17):3874–3876. [PubMed] [Google Scholar]
  17. Dumaz N., Drougard C., Sarasin A., Daya-Grosjean L. Specific UV-induced mutation spectrum in the p53 gene of skin tumors from DNA-repair-deficient xeroderma pigmentosum patients. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10529–10533. doi: 10.1073/pnas.90.22.10529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gentil A., Renault G., Madzak C., Margot A., Cabral-Neto J. B., Vasseur J. J., Rayner B., Imbach J. L., Sarasin A. Mutagenic properties of a unique abasic site in mammalian cells. Biochem Biophys Res Commun. 1990 Dec 14;173(2):704–710. doi: 10.1016/s0006-291x(05)80092-0. [DOI] [PubMed] [Google Scholar]
  19. Greene M. H. Is cisplatin a human carcinogen? J Natl Cancer Inst. 1992 Mar 4;84(5):306–312. doi: 10.1093/jnci/84.5.306. [DOI] [PubMed] [Google Scholar]
  20. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  21. Hennings H., Shores R. A., Poirier M. C., Reed E., Tarone R. E., Yuspa S. H. Enhanced malignant conversion of benign mouse skin tumors by cisplatin. J Natl Cancer Inst. 1990 May 16;82(10):836–840. doi: 10.1093/jnci/82.10.836. [DOI] [PubMed] [Google Scholar]
  22. Herman F., Kozelka J., Stoven V., Guittet E., Girault J. P., Huynh-Dinh T., Igolen J., Lallemand J. Y., Chottard J. C. A d(GpG)-platinated decanucleotide duplex is kinked. An extended NMR and molecular mechanics study. Eur J Biochem. 1990 Nov 26;194(1):119–133. doi: 10.1111/j.1432-1033.1990.tb19435.x. [DOI] [PubMed] [Google Scholar]
  23. Hoffmann J. S., Fry M., Ji J., Williams K. J., Loeb L. A. Codons 12 and 13 of H-ras protooncogene interrupt the progression of DNA synthesis catalyzed by DNA polymerase alpha. Cancer Res. 1993 Jun 15;53(12):2895–2900. [PubMed] [Google Scholar]
  24. Hoffmann J. S., Johnson N. P., Villani G. Conversion of monofunctional DNA adducts of cis-diamminedichloroplatinum (II) to bifunctional lesions. Effect on the in vitro replication of single-stranded DNA by Escherichia coli DNA polymerase I and eukaryotic DNA polymerases alpha. J Biol Chem. 1989 Sep 5;264(25):15130–15135. [PubMed] [Google Scholar]
  25. Huang L., Turchi J. J., Wahl A. F., Bambara R. A. Activity of calf thymus DNA helicase E on cis-diamminedichloroplatinum (II)-damaged DNA. J Biol Chem. 1993 Dec 15;268(35):26731–26737. [PubMed] [Google Scholar]
  26. Huang L., Turchi J. J., Wahl A. F., Bambara R. A. Effects of the anticancer drug cis-diamminedichloroplatinum(II) on the activities of calf thymus DNA polymerase epsilon. Biochemistry. 1993 Jan 26;32(3):841–848. doi: 10.1021/bi00054a015. [DOI] [PubMed] [Google Scholar]
  27. Isonishi S., Hom D. K., Thiebaut F. B., Mann S. C., Andrews P. A., Basu A., Lazo J. S., Eastman A., Howell S. B. Expression of the c-Ha-ras oncogene in mouse NIH 3T3 cells induces resistance to cisplatin. Cancer Res. 1991 Nov 1;51(21):5903–5909. [PubMed] [Google Scholar]
  28. Loehrer P. J., Einhorn L. H. Drugs five years later. Cisplatin. Ann Intern Med. 1984 May;100(5):704–713. doi: 10.7326/0003-4819-100-5-704. [DOI] [PubMed] [Google Scholar]
  29. Madzak C., Sarasin A. Conversion of a single-stranded simian virus 40 (SV40)-based shuttle vector to its double-stranded form does not require the SV40 T antigen in monkey cells. J Gen Virol. 1991 Dec;72(Pt 12):3091–3093. doi: 10.1099/0022-1317-72-12-3091. [DOI] [PubMed] [Google Scholar]
  30. Marrot L., Leng M. Chemical probes of the conformation of DNA modified by cis-diamminedichloroplatinum(II). Biochemistry. 1989 Feb 21;28(4):1454–1461. doi: 10.1021/bi00430a005. [DOI] [PubMed] [Google Scholar]
  31. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  32. Mis J. R., Kunz B. A. Analysis of mutations induced in the SUP4-o gene of Saccharomyces cerevisiae by cis-diammine dichloroplatinum(II). Carcinogenesis. 1990 Apr;11(4):633–638. doi: 10.1093/carcin/11.4.633. [DOI] [PubMed] [Google Scholar]
  33. Murray V., Motyka H., England P. R., Wickham G., Lee H. H., Denny W. A., McFadyen W. D. The use of Taq DNA polymerase to determine the sequence specificity of DNA damage caused by cis-diamminedichloroplatinum(II), acridine-tethered platinum(II) diammine complexes or two analogues. J Biol Chem. 1992 Sep 15;267(26):18805–18809. [PubMed] [Google Scholar]
  34. Naegeli H., Bardwell L., Friedberg E. C. Inhibition of Rad3 DNA helicase activity by DNA adducts and abasic sites: implications for the role of a DNA helicase in damage-specific incision of DNA. Biochemistry. 1993 Jan 19;32(2):613–621. doi: 10.1021/bi00053a029. [DOI] [PubMed] [Google Scholar]
  35. Naser L. J., Pinto A. L., Lippard S. J., Essigmann J. M. Chemical and biological studies of the major DNA adduct of cis-diamminedichloroplatinum(II), cis-[Pt(NH3)2(d(GpG]], built into a specific site in a viral genome. Biochemistry. 1988 Jun 14;27(12):4357–4367. doi: 10.1021/bi00412a024. [DOI] [PubMed] [Google Scholar]
  36. Pillaire M. J., Villani G., Hoffmann J. S., Mazard A. M., Defais M. Characterization and localization of cis-diamminedichloro-platinum(II) adducts on a purified oligonucleotide containing the codons 12 and 13 of H-ras proto-oncogene. Nucleic Acids Res. 1992 Dec 25;20(24):6473–6479. doi: 10.1093/nar/20.24.6473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Seidman M. The development of transient SV40 based shuttle vectors for mutagenesis studies: problems and solutions. Mutat Res. 1989 Mar-May;220(2-3):55–60. doi: 10.1016/0165-1110(89)90010-9. [DOI] [PubMed] [Google Scholar]
  38. Sklar M. D. Increased resistance to cis-diamminedichloroplatinum(II) in NIH 3T3 cells transformed by ras oncogenes. Cancer Res. 1988 Feb 15;48(4):793–797. [PubMed] [Google Scholar]
  39. Stary A., Sarasin A. Simian virus 40 (SV40) large T antigen-dependent amplification of an Epstein-Barr virus-SV40 hybrid shuttle vector integrated into the human HeLa cell genome. J Gen Virol. 1992 Jul;73(Pt 7):1679–1685. doi: 10.1099/0022-1317-73-7-1679. [DOI] [PubMed] [Google Scholar]
  40. Strauss B. S. The 'A rule' of mutagen specificity: a consequence of DNA polymerase bypass of non-instructional lesions? Bioessays. 1991 Feb;13(2):79–84. doi: 10.1002/bies.950130206. [DOI] [PubMed] [Google Scholar]
  41. Szymkowski D. E., Yarema K., Essigmann J. M., Lippard S. J., Wood R. D. An intrastrand d(GpG) platinum crosslink in duplex M13 DNA is refractory to repair by human cell extracts. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10772–10776. doi: 10.1073/pnas.89.22.10772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Villani G., Cazaux C., Pillaire M. J., Boehmer P. Effects of a single intrastrand d(GpG) platinum adduct on the strand separating activity of the Escherichia coli proteins RecB and RecA. FEBS Lett. 1993 Oct 25;333(1-2):89–95. doi: 10.1016/0014-5793(93)80380-d. [DOI] [PubMed] [Google Scholar]
  43. Villani G., Hübscher U., Butour J. L. Sites of termination of in vitro DNA synthesis on cis-diamminedichloroplatinum(II) treated single-stranded DNA: a comparison between E. coli DNA polymerase I and eucaryotic DNA polymerases alpha. Nucleic Acids Res. 1988 May 25;16(10):4407–4418. doi: 10.1093/nar/16.10.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. de Boer J. G., Glickman B. W. Sequence specificity of mutation induced by the anti-tumor drug cisplatin in the CHO aprt gene. Carcinogenesis. 1989 Aug;10(8):1363–1367. doi: 10.1093/carcin/10.8.1363. [DOI] [PubMed] [Google Scholar]
  45. den Hartog J. H., Altona C., van Boom J. H., van der Marel G. A., Haasnoot C. A., Reedijk J. cis-diamminedichloroplatinum(II) induced distortion of a single and double stranded deoxydecanucleosidenonaphosphate studied by nuclear magnetic resonance. J Biomol Struct Dyn. 1985 Jun;2(6):1137–1155. doi: 10.1080/07391102.1985.10507629. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES