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. 1996 May 1;315(Pt 3):821–825. doi: 10.1042/bj3150821

Chinese hamster fibroblasts overexpressing CuZn-superoxide dismutase undergo a global reduction in antioxidants and an increasing sensitivity of DNA to oxidative damage.

H D Teixeira 1, R Meneghini 1
PMCID: PMC1217280  PMID: 8645163

Abstract

Transfection of a CuZn-superoxide dismutase (SOD) expression vector into V79 Chinese hamster cells produced clones in which CuZn-SOD activities were 2.2-3.5-fold higher than in the parental cells. An overall moderate reduction of glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase activities and of both GSSG and total glutathione levels was found. In one particular clone the catalase activity was also reduced. The pro-oxidant status established by the lower level of antioxidant defence rendered the SOD-overexpressing cells more sensitive to the production of 8-oxo-2'-deoxyguanosine by hydrogen peroxide. The data are discussed in terms of a model resembling the bacterial sox RS system.

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Selected References

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

  1. Akerboom T. P., Sies H. Assay of glutathione, glutathione disulfide, and glutathione mixed disulfides in biological samples. Methods Enzymol. 1981;77:373–382. doi: 10.1016/s0076-6879(81)77050-2. [DOI] [PubMed] [Google Scholar]
  2. Amstad P., Moret R., Cerutti P. Glutathione peroxidase compensates for the hypersensitivity of Cu,Zn-superoxide dismutase overproducers to oxidant stress. J Biol Chem. 1994 Jan 21;269(3):1606–1609. [PubMed] [Google Scholar]
  3. Amstad P., Peskin A., Shah G., Mirault M. E., Moret R., Zbinden I., Cerutti P. The balance between Cu,Zn-superoxide dismutase and catalase affects the sensitivity of mouse epidermal cells to oxidative stress. Biochemistry. 1991 Sep 24;30(38):9305–9313. doi: 10.1021/bi00102a024. [DOI] [PubMed] [Google Scholar]
  4. Avraham K. B., Schickler M., Sapoznikov D., Yarom R., Groner Y. Down's syndrome: abnormal neuromuscular junction in tongue of transgenic mice with elevated levels of human Cu/Zn-superoxide dismutase. Cell. 1988 Sep 9;54(6):823–829. doi: 10.1016/s0092-8674(88)91153-1. [DOI] [PubMed] [Google Scholar]
  5. Bertoncini C. R., Meneghini R. DNA strand breaks produced by oxidative stress in mammalian cells exhibit 3'-phosphoglycolate termini. Nucleic Acids Res. 1995 Aug 11;23(15):2995–3002. doi: 10.1093/nar/23.15.2995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Birnboim H. C. Rapid extraction of high molecular weight RNA from cultured cells and granulocytes for Northern analysis. Nucleic Acids Res. 1988 Feb 25;16(4):1487–1497. doi: 10.1093/nar/16.4.1487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  8. Carlioz A., Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. doi: 10.1002/j.1460-2075.1986.tb04256.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Demple B. Regulation of bacterial oxidative stress genes. Annu Rev Genet. 1991;25:315–337. doi: 10.1146/annurev.ge.25.120191.001531. [DOI] [PubMed] [Google Scholar]
  10. Deng H. X., Hentati A., Tainer J. A., Iqbal Z., Cayabyab A., Hung W. Y., Getzoff E. D., Hu P., Herzfeldt B., Roos R. P. Amyotrophic lateral sclerosis and structural defects in Cu,Zn superoxide dismutase. Science. 1993 Aug 20;261(5124):1047–1051. doi: 10.1126/science.8351519. [DOI] [PubMed] [Google Scholar]
  11. Elroy-Stein O., Bernstein Y., Groner Y. Overproduction of human Cu/Zn-superoxide dismutase in transfected cells: extenuation of paraquat-mediated cytotoxicity and enhancement of lipid peroxidation. EMBO J. 1986 Mar;5(3):615–622. doi: 10.1002/j.1460-2075.1986.tb04255.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Elroy-Stein O., Groner Y. Impaired neurotransmitter uptake in PC12 cells overexpressing human Cu/Zn-superoxide dismutase--implication for gene dosage effects in Down syndrome. Cell. 1988 Jan 29;52(2):259–267. doi: 10.1016/0092-8674(88)90515-6. [DOI] [PubMed] [Google Scholar]
  13. Epstein C. J., Avraham K. B., Lovett M., Smith S., Elroy-Stein O., Rotman G., Bry C., Groner Y. Transgenic mice with increased Cu/Zn-superoxide dismutase activity: animal model of dosage effects in Down syndrome. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8044–8048. doi: 10.1073/pnas.84.22.8044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Farr S. B., Kogoma T. Oxidative stress responses in Escherichia coli and Salmonella typhimurium. Microbiol Rev. 1991 Dec;55(4):561–585. doi: 10.1128/mr.55.4.561-585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Flohé L., Günzler W. A. Assays of glutathione peroxidase. Methods Enzymol. 1984;105:114–121. doi: 10.1016/s0076-6879(84)05015-1. [DOI] [PubMed] [Google Scholar]
  16. GLOCK G. E., McLEAN P. Further studies on the properties and assay of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase of rat liver. Biochem J. 1953 Oct;55(3):400–408. doi: 10.1042/bj0550400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Griffith O. W. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem. 1980 Jul 15;106(1):207–212. doi: 10.1016/0003-2697(80)90139-6. [DOI] [PubMed] [Google Scholar]
  18. Hoffmann M. E., Mello-Filho A. C., Meneghini R. Correlation between cytotoxic effect of hydrogen peroxide and the yield of DNA strand breaks in cells of different species. Biochim Biophys Acta. 1984 Apr 5;781(3):234–238. doi: 10.1016/0167-4781(84)90088-5. [DOI] [PubMed] [Google Scholar]
  19. Huang T. T., Carlson E. J., Leadon S. A., Epstein C. J. Relationship of resistance to oxygen free radicals to CuZn-superoxide dismutase activity in transgenic, transfected, and trisomic cells. FASEB J. 1992 Feb 1;6(3):903–910. doi: 10.1096/fasebj.6.3.1740238. [DOI] [PubMed] [Google Scholar]
  20. Kelner M. J., Bagnell R. Alteration of endogenous glutathione peroxidase, manganese superoxide dismutase, and glutathione transferase activity in cells transfected with a copper-zinc superoxide dismutase expression vector. Explanation for variations in paraquat resistance. J Biol Chem. 1990 Jul 5;265(19):10872–10875. [PubMed] [Google Scholar]
  21. Kelner M. J., Bagnell R., Montoya M., Estes L., Uglik S. F., Cerutti P. Transfection with human copper-zinc superoxide dismutase induces bidirectional alterations in other antioxidant enzymes, proteins, growth factor response, and paraquat resistance. Free Radic Biol Med. 1995 Mar;18(3):497–506. doi: 10.1016/0891-5849(94)00167-i. [DOI] [PubMed] [Google Scholar]
  22. Li Z., Demple B. SoxS, an activator of superoxide stress genes in Escherichia coli. Purification and interaction with DNA. J Biol Chem. 1994 Jul 15;269(28):18371–18377. [PubMed] [Google Scholar]
  23. Liochev S. I., Fridovich I. Effects of overproduction of superoxide dismutase on the toxicity of paraquat toward Escherichia coli. J Biol Chem. 1991 May 15;266(14):8747–8750. [PubMed] [Google Scholar]
  24. Liochev S. I., Fridovich I. The role of O2.- in the production of HO.: in vitro and in vivo. Free Radic Biol Med. 1994 Jan;16(1):29–33. doi: 10.1016/0891-5849(94)90239-9. [DOI] [PubMed] [Google Scholar]
  25. Martins E. A., Robalinho R. L., Meneghini R. Oxidative stress induces activation of a cytosolic protein responsible for control of iron uptake. Arch Biochem Biophys. 1995 Jan 10;316(1):128–134. doi: 10.1006/abbi.1995.1019. [DOI] [PubMed] [Google Scholar]
  26. Minc-Golomb D., Knobler H., Groner Y. Gene dosage of CuZnSOD and Down's syndrome: diminished prostaglandin synthesis in human trisomy 21, transfected cells and transgenic mice. EMBO J. 1991 Aug;10(8):2119–2124. doi: 10.1002/j.1460-2075.1991.tb07745.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nose K., Shibanuma M., Kikuchi K., Kageyama H., Sakiyama S., Kuroki T. Transcriptional activation of early-response genes by hydrogen peroxide in a mouse osteoblastic cell line. Eur J Biochem. 1991 Oct 1;201(1):99–106. doi: 10.1111/j.1432-1033.1991.tb16261.x. [DOI] [PubMed] [Google Scholar]
  28. Oyanagui Y. Reevaluation of assay methods and establishment of kit for superoxide dismutase activity. Anal Biochem. 1984 Nov 1;142(2):290–296. doi: 10.1016/0003-2697(84)90467-6. [DOI] [PubMed] [Google Scholar]
  29. Pantopoulos K., Hentze M. W. Rapid responses to oxidative stress mediated by iron regulatory protein. EMBO J. 1995 Jun 15;14(12):2917–2924. doi: 10.1002/j.1460-2075.1995.tb07291.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Przedborski S., Jackson-Lewis V., Kostic V., Carlson E., Epstein C. J., Cadet J. L. Superoxide dismutase, catalase, and glutathione peroxidase activities in copper/zinc-superoxide dismutase transgenic mice. J Neurochem. 1992 May;58(5):1760–1767. doi: 10.1111/j.1471-4159.1992.tb10051.x. [DOI] [PubMed] [Google Scholar]
  31. Rao G. N., Berk B. C. Active oxygen species stimulate vascular smooth muscle cell growth and proto-oncogene expression. Circ Res. 1992 Mar;70(3):593–599. doi: 10.1161/01.res.70.3.593. [DOI] [PubMed] [Google Scholar]
  32. Rosen D. R., Siddique T., Patterson D., Figlewicz D. A., Sapp P., Hentati A., Donaldson D., Goto J., O'Regan J. P., Deng H. X. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature. 1993 Mar 4;362(6415):59–62. doi: 10.1038/362059a0. [DOI] [PubMed] [Google Scholar]
  33. Schmidt K. N., Amstad P., Cerutti P., Baeuerle P. A. The roles of hydrogen peroxide and superoxide as messengers in the activation of transcription factor NF-kappa B. Chem Biol. 1995 Jan;2(1):13–22. doi: 10.1016/1074-5521(95)90076-4. [DOI] [PubMed] [Google Scholar]
  34. Scott M. D., Meshnick S. R., Eaton J. W. Superoxide dismutase amplifies organismal sensitivity to ionizing radiation. J Biol Chem. 1989 Feb 15;264(5):2498–2501. [PubMed] [Google Scholar]
  35. Scott M. D., Meshnick S. R., Eaton J. W. Superoxide dismutase-rich bacteria. Paradoxical increase in oxidant toxicity. J Biol Chem. 1987 Mar 15;262(8):3640–3645. [PubMed] [Google Scholar]
  36. Shibanuma M., Kuroki T., Nose K. Stimulation by hydrogen peroxide of DNA synthesis, competence family gene expression and phosphorylation of a specific protein in quiescent Balb/3T3 cells. Oncogene. 1990 Jul;5(7):1025–1032. [PubMed] [Google Scholar]
  37. Shigenaga M. K., Park J. W., Cundy K. C., Gimeno C. J., Ames B. N. In vivo oxidative DNA damage: measurement of 8-hydroxy-2'-deoxyguanosine in DNA and urine by high-performance liquid chromatography with electrochemical detection. Methods Enzymol. 1990;186:521–530. doi: 10.1016/0076-6879(90)86146-m. [DOI] [PubMed] [Google Scholar]
  38. Sies H., Koch O. R., Martino E., Boveris A. Increased biliary glutathione disulfide release in chronically ethanol-treated rats. FEBS Lett. 1979 Jul 15;103(2):287–290. doi: 10.1016/0014-5793(79)81346-0. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Sundaresan M., Yu Z. X., Ferrans V. J., Irani K., Finkel T. Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science. 1995 Oct 13;270(5234):296–299. doi: 10.1126/science.270.5234.296. [DOI] [PubMed] [Google Scholar]
  41. Wong G. H., Elwell J. H., Oberley L. W., Goeddel D. V. Manganous superoxide dismutase is essential for cellular resistance to cytotoxicity of tumor necrosis factor. Cell. 1989 Sep 8;58(5):923–931. doi: 10.1016/0092-8674(89)90944-6. [DOI] [PubMed] [Google Scholar]

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