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. 1989 Oct 25;17(20):8301–8312. doi: 10.1093/nar/17.20.8301

The spectrum of mutations generated by passage of a hydrogen peroxide damaged shuttle vector plasmid through a mammalian host.

E C Moraes 1, S M Keyse 1, M Pidoux 1, R M Tyrrell 1
PMCID: PMC334965  PMID: 2682525

Abstract

Treatment of a plasmid shuttle vector (pZ189) with a combination of hydrogen peroxide and a ferric iron/EDTA complex prior to transfection and passage in simian (CV-1) cells increases the frequency of mutations at the supF locus by up to 60-fold over the spontaneous background. This increase in mutation frequency is abolished when the inhibitors desferrioxamine, superoxide dismutase, catalase or dimethyl sulfoxide are included in the initial reaction or when the iron/EDTA complex is omitted, a strong indication that the premutagenic damage arises as a result of direct attack by hydroxyl radical generated in a superoxide driven Fenton reaction. DNA sequence analysis of the mutated plasmids shows that 1) Deletions occuring in combination with base-substitutions arise in 22.5 percent of the induced mutants compared with only 3 percent of spontaneous mutants 2) Sixty percent of all induced deletion mutations involve the loss of a single base and 77 percent of these (20 out of 26) occur at two adenine-containing sites 3) The base-change spectrum of mutants arising in the treated plasmid population is marked by the predominance of mutants containing a single base-change and by an increase in changes at AT base pairs. These results provide direct information concerning the nature of mutations arising in mammalian cells as a result of hydroxyl radical mediated DNA damage.

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Selected References

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

  1. Farabaugh P. J., Schmeissner U., Hofer M., Miller J. H. Genetic studies of the lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol. 1978 Dec 25;126(4):847–857. doi: 10.1016/0022-2836(78)90023-2. [DOI] [PubMed] [Google Scholar]
  2. Gutteridge J. M. Superoxide dismutase inhibits the superoxide-driven Fenton reaction at two different levels. Implications for a wider protective role. FEBS Lett. 1985 Jun 3;185(1):19–23. doi: 10.1016/0014-5793(85)80732-8. [DOI] [PubMed] [Google Scholar]
  3. Hauser J., Levine A. S., Dixon K. Unique pattern of point mutations arising after gene transfer into mammalian cells. EMBO J. 1987 Jan;6(1):63–67. doi: 10.1002/j.1460-2075.1987.tb04719.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hauser J., Seidman M. M., Sidur K., Dixon K. Sequence specificity of point mutations induced during passage of a UV-irradiated shuttle vector plasmid in monkey cells. Mol Cell Biol. 1986 Jan;6(1):277–285. doi: 10.1128/mcb.6.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  6. Imlay J. A., Linn S. Mutagenesis and stress responses induced in Escherichia coli by hydrogen peroxide. J Bacteriol. 1987 Jul;169(7):2967–2976. doi: 10.1128/jb.169.7.2967-2976.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Keyse S. M., Amaudruz F., Tyrrell R. M. Determination of the spectrum of mutations induced by defined-wavelength solar UVB (313-nm) radiation in mammalian cells by use of a shuttle vector. Mol Cell Biol. 1988 Dec;8(12):5425–5431. doi: 10.1128/mcb.8.12.5425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Levin D. E., Hollstein M., Christman M. F., Schwiers E. A., Ames B. N. A new Salmonella tester strain (TA102) with A X T base pairs at the site of mutation detects oxidative mutagens. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7445–7449. doi: 10.1073/pnas.79.23.7445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Loeb L. A., James E. A., Waltersdorph A. M., Klebanoff S. J. Mutagenesis by the autoxidation of iron with isolated DNA. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3918–3922. doi: 10.1073/pnas.85.11.3918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McCormick J. P., Fischer J. R., Pachlatko J. P., Eisenstark A. Characterization of a cell-lethal product from the photooxidation of tryptophan: hydrogen peroxide. Science. 1976 Feb 6;191(4226):468–469. doi: 10.1126/science.1108203. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. McMaster G. K., Beard P., Engers H. D., Hirt B. Characterization of an immunosuppressive parvovirus related to the minute virus of mice. J Virol. 1981 Apr;38(1):317–326. doi: 10.1128/jvi.38.1.317-326.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mello Filho A. C., Hoffmann M. E., Meneghini R. Cell killing and DNA damage by hydrogen peroxide are mediated by intracellular iron. Biochem J. 1984 Feb 15;218(1):273–275. doi: 10.1042/bj2180273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miller J. H. Mutagenic specificity of ultraviolet light. J Mol Biol. 1985 Mar 5;182(1):45–65. doi: 10.1016/0022-2836(85)90026-9. [DOI] [PubMed] [Google Scholar]
  15. Phear G., Nalbantoglu J., Meuth M. Next-nucleotide effects in mutations driven by DNA precursor pool imbalances at the aprt locus of Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4450–4454. doi: 10.1073/pnas.84.13.4450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rajagopalan S., Politi P. M., Sinha B. K., Myers C. E. Adriamycin-induced free radical formation in the perfused rat heart: implications for cardiotoxicity. Cancer Res. 1988 Sep 1;48(17):4766–4769. [PubMed] [Google Scholar]
  17. Repine J. E., Pfenninger O. W., Talmage D. W., Berger E. M., Pettijohn D. E. Dimethyl sulfoxide prevents DNA nicking mediated by ionizing radiation or iron/hydrogen peroxide-generated hydroxyl radical. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1001–1003. doi: 10.1073/pnas.78.2.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rhaese H. J., Freese E. Chemical analysis of DNA alterations. I. Base liberation and backbone breakage of DNA and oligodeoxyadenylic acid induced by hydrogen peroxide and hydroxylamine. Biochim Biophys Acta. 1968 Feb 26;155(2):476–490. [PubMed] [Google Scholar]
  19. Sagripanti J. L., Kraemer K. H. Site-specific oxidative DNA damage at polyguanosines produced by copper plus hydrogen peroxide. J Biol Chem. 1989 Jan 25;264(3):1729–1734. [PubMed] [Google Scholar]
  20. Sahli R., McMaster G. K., Hirt B. DNA sequence comparison between two tissue-specific variants of the autonomous parvovirus, minute virus of mice. Nucleic Acids Res. 1985 May 24;13(10):3617–3633. doi: 10.1093/nar/13.10.3617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Seidman M. M., Dixon K., Razzaque A., Zagursky R. J., Berman M. L. A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells. Gene. 1985;38(1-3):233–237. doi: 10.1016/0378-1119(85)90222-7. [DOI] [PubMed] [Google Scholar]
  22. Starke P. E., Farber J. L. Ferric iron and superoxide ions are required for the killing of cultured hepatocytes by hydrogen peroxide. Evidence for the participation of hydroxyl radicals formed by an iron-catalyzed Haber-Weiss reaction. J Biol Chem. 1985 Aug 25;260(18):10099–10104. [PubMed] [Google Scholar]
  23. Streisinger G., Okada Y., Emrich J., Newton J., Tsugita A., Terzaghi E., Inouye M. Frameshift mutations and the genetic code. This paper is dedicated to Professor Theodosius Dobzhansky on the occasion of his 66th birthday. Cold Spring Harb Symp Quant Biol. 1966;31:77–84. doi: 10.1101/sqb.1966.031.01.014. [DOI] [PubMed] [Google Scholar]
  24. Yamafuji K., Uchida Y. Liberation of adenine from deoxyribonucleic acid by hydrogen peroxide. Nature. 1966 Jan 15;209(5020):301–302. doi: 10.1038/209301a0. [DOI] [PubMed] [Google Scholar]
  25. Ziegler-Skylakakis K., Andrae U. Mutagenicity of hydrogen peroxide in V79 Chinese hamster cells. Mutat Res. 1987 Sep;192(1):65–67. doi: 10.1016/0165-7992(87)90127-8. [DOI] [PubMed] [Google Scholar]

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