Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1986 Mar;5(3):615–622. doi: 10.1002/j.1460-2075.1986.tb04255.x

Overproduction of human Cu/Zn-superoxide dismutase in transfected cells: extenuation of paraquat-mediated cytotoxicity and enhancement of lipid peroxidation.

O Elroy-Stein, Y Bernstein, Y Groner
PMCID: PMC1166807  PMID: 3011416

Abstract

The 'housekeeping' enzyme Cu/Zn-superoxide dismutase (SOD-1) is encoded by a gene residing on human chromosome 21, at the region 21q22 known to be involved in Down's syndrome. The SOD-1 gene and the SOD-1 cDNA were introduced into mouse L-cells and human HeLa cells, respectively as part of recombinant plasmids containing the neoR selectable marker. Human and mouse transformants were obtained that expressed elevated levels (up to 6-fold) of authentic, enzymatically active human SOD-1. This enabled us to examine the consequences of hSOD-1 gene dosage, apart from gene dosage effects contributed by other genes residing on chromosome 21. Human and mouse cell clones that overproduce the hSOD-1 had altered properties; they were more resistant to paraquat than the parental cells and showed an increase in lipid peroxidation. The data are consistent with the possibility that gene dosage of hSOD-1 contributes to some of the clinical symptoms associated with Down's syndrome.

Full text

PDF
615

Images in this article

616Fig. 2.
on p.616
617Fig. 3.
on p.617
618Fig. 4.
on p.618
618Fig. 5.
on p.618

Selected References

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

  1. Badwey J. A., Karnovsky M. L. Active oxygen species and the functions of phagocytic leukocytes. Annu Rev Biochem. 1980;49:695–726. doi: 10.1146/annurev.bi.49.070180.003403. [DOI] [PubMed] [Google Scholar]
  2. Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Boehme D. H., Kosecki R., Carson S., Stern F., Marks N. Lipoperoxidation in human and rat brain tissue: developmental and regional studies. Brain Res. 1977 Nov 4;136(1):11–21. doi: 10.1016/0006-8993(77)90127-5. [DOI] [PubMed] [Google Scholar]
  5. Brooksbank B. W., Balazs R. Superoxide dismutase, glutathione peroxidase and lipoperoxidation in Down's syndrome fetal brain. Brain Res. 1984 Sep;318(1):37–44. doi: 10.1016/0165-3806(84)90060-9. [DOI] [PubMed] [Google Scholar]
  6. Brooksbank B. W., Martinez M., Balazs R. Altered composition of polyunsaturated fatty acyl groups in phosphoglycerides of Down's syndrome fetal brain. J Neurochem. 1985 Mar;44(3):869–874. doi: 10.1111/j.1471-4159.1985.tb12896.x. [DOI] [PubMed] [Google Scholar]
  7. Canaani D., Berg P. Regulated expression of human interferon beta 1 gene after transduction into cultured mouse and rabbit cells. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5166–5170. doi: 10.1073/pnas.79.17.5166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DAHLE L. K., HILL E. G., HOLMAN R. T. The thiobarbituric acid reaction and the autoxidations of polyunsaturated fatty acid methyl esters. Arch Biochem Biophys. 1962 Aug;98:253–261. doi: 10.1016/0003-9861(62)90181-9. [DOI] [PubMed] [Google Scholar]
  9. Elstner E. F., Heupel A. Inhibition of nitrite formation from hydroxylammoniumchloride: a simple assay for superoxide dismutase. Anal Biochem. 1976 Feb;70(2):616–620. doi: 10.1016/0003-2697(76)90488-7. [DOI] [PubMed] [Google Scholar]
  10. Epstein C. J., Epstein L. B., Cox D. R., Weil J. Functional implications of gene dosage effects in trisomy 21. Hum Genet Suppl. 1981;2:155–172. doi: 10.1007/978-3-642-68006-9_12. [DOI] [PubMed] [Google Scholar]
  11. Epstein C. J., Epstein L. B., Weil J., Cox D. R. Trisomy 21: mechanisms and models. Ann N Y Acad Sci. 1982;396:107–118. doi: 10.1111/j.1749-6632.1982.tb26847.x. [DOI] [PubMed] [Google Scholar]
  12. Farrington J. A., Ebert M., Land E. J., Fletcher K. Bipyridylium quaternary salts and related compounds. V. Pulse radiolysis studies of the reaction of paraquat radical with oxygen. Implications for the mode of action of bipyridyl herbicides. Biochim Biophys Acta. 1973 Sep 26;314(3):372–381. doi: 10.1016/0005-2728(73)90121-7. [DOI] [PubMed] [Google Scholar]
  13. Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
  14. Frischer H., Chu L. K., Ahmad T., Justice P., Smith G. F. Superoxide dismutase and glutathione peroxidase abnormalities in erythrocytes and lymphoid cells in Down syndrome. Prog Clin Biol Res. 1981;55:269–289. [PubMed] [Google Scholar]
  15. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  16. Hassan H. M., Fridovich I. Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. Arch Biochem Biophys. 1979 Sep;196(2):385–395. doi: 10.1016/0003-9861(79)90289-3. [DOI] [PubMed] [Google Scholar]
  17. Hassan H. M., Fridovich I. Regulation of the synthesis of superoxide dismutase in Escherichia coli. Induction by methyl viologen. J Biol Chem. 1977 Nov 10;252(21):7667–7672. [PubMed] [Google Scholar]
  18. Hassold T. J., Jacobs P. A. Trisomy in man. Annu Rev Genet. 1984;18:69–97. doi: 10.1146/annurev.ge.18.120184.000441. [DOI] [PubMed] [Google Scholar]
  19. Kanner J., Harel S. Initiation of membranal lipid peroxidation by activated metmyoglobin and methemoglobin. Arch Biochem Biophys. 1985 Mar;237(2):314–321. doi: 10.1016/0003-9861(85)90282-6. [DOI] [PubMed] [Google Scholar]
  20. Kedziora J., Lukaszewicz R., Koter M., Bartosz G., Pawłowska B., Aitkin D. Red blood cell glutathione peroxidase in simple trisomy 21 and translocation 21/22. Experientia. 1982 May 15;38(5):543–544. doi: 10.1007/BF02327040. [DOI] [PubMed] [Google Scholar]
  21. LEJEUNE J., GAUTIER M., TURPIN R. Etude des chromosomes somatiques de neuf enfants mongoliens. C R Hebd Seances Acad Sci. 1959 Mar 16;248(11):1721–1722. [PubMed] [Google Scholar]
  22. Levanon D., Lieman-Hurwitz J., Dafni N., Wigderson M., Sherman L., Bernstein Y., Laver-Rudich Z., Danciger E., Stein O., Groner Y. Architecture and anatomy of the chromosomal locus in human chromosome 21 encoding the Cu/Zn superoxide dismutase. EMBO J. 1985 Jan;4(1):77–84. doi: 10.1002/j.1460-2075.1985.tb02320.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lieman-Hurwitz J., Dafni N., Lavie V., Groner Y. Human cytoplasmic superoxide dismutase cDNA clone: a probe for studying the molecular biology of Down syndrome. Proc Natl Acad Sci U S A. 1982 May;79(9):2808–2811. doi: 10.1073/pnas.79.9.2808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
  25. Malmström B. G. Enzymology of oxygen. Annu Rev Biochem. 1982;51:21–59. doi: 10.1146/annurev.bi.51.070182.000321. [DOI] [PubMed] [Google Scholar]
  26. McCoy E. E., Enns L. Sodium transport ouabain binding, and (Na+/K+)-ATPase activity in Down's syndrome platelets. Pediatr Res. 1978 Jun;12(6):685–689. doi: 10.1203/00006450-197806000-00001. [DOI] [PubMed] [Google Scholar]
  27. Moody C. S., Hassan H. M. Mutagenicity of oxygen free radicals. Proc Natl Acad Sci U S A. 1982 May;79(9):2855–2859. doi: 10.1073/pnas.79.9.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mulligan R. C., Berg P. Expression of a bacterial gene in mammalian cells. Science. 1980 Sep 19;209(4463):1422–1427. doi: 10.1126/science.6251549. [DOI] [PubMed] [Google Scholar]
  29. Niehaus W. G., Jr, Samuelsson B. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur J Biochem. 1968 Oct 17;6(1):126–130. doi: 10.1111/j.1432-1033.1968.tb00428.x. [DOI] [PubMed] [Google Scholar]
  30. Pantelakis S. N., Karaklis A. G., Alexiou D., Vardas E., Valaes T. Red cell enzymes in trisomy 21. Am J Hum Genet. 1970 Mar;22(2):184–193. [PMC free article] [PubMed] [Google Scholar]
  31. Patterson D., Jones C., Scoggin C., Miller Y. E., Graw S. Somatic cell genetic approaches to Down's syndrome. Ann N Y Acad Sci. 1982;396:69–81. doi: 10.1111/j.1749-6632.1982.tb26844.x. [DOI] [PubMed] [Google Scholar]
  32. Pryor W. A., Stanley J. P., Blair E. Autoxidation of polyunsaturated fatty acids: II. A suggested mechanism for the formation of TBA-reactive materials from prostaglandin-like endoperoxides. Lipids. 1976 May;11(5):370–379. doi: 10.1007/BF02532843. [DOI] [PubMed] [Google Scholar]
  33. Scoggin C. H., Patterson D. Down's syndrome as a model disease. Arch Intern Med. 1982 Mar;142(3):462–464. [PubMed] [Google Scholar]
  34. Sherman L., Dafni N., Lieman-Hurwitz J., Groner Y. Nucleotide sequence and expression of human chromosome 21-encoded superoxide dismutase mRNA. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5465–5469. doi: 10.1073/pnas.80.18.5465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sherman L., Levanon D., Lieman-Hurwitz J., Dafni N., Groner Y. Human Cu/Zn superoxide dismutase gene: molecular characterization of its two mRNA species. Nucleic Acids Res. 1984 Dec 21;12(24):9349–9365. doi: 10.1093/nar/12.24.9349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sinet P. M. Metabolism of oxygen derivatives in down's syndrome. Ann N Y Acad Sci. 1982;396:83–94. doi: 10.1111/j.1749-6632.1982.tb26845.x. [DOI] [PubMed] [Google Scholar]
  37. Sinet P. M., Michelson A. M., Bazin A., Lejeune J., Jerome H. Increase in glutathione peroxidase activity in erythrocytes from trisomy 21 subjects. Biochem Biophys Res Commun. 1975 Dec 1;67(3):910–915. doi: 10.1016/0006-291x(75)90763-9. [DOI] [PubMed] [Google Scholar]
  38. Smith G. F., Warren S. T. The biology of Down syndrome. Ann N Y Acad Sci. 1985;450:1–9. doi: 10.1111/j.1749-6632.1985.tb21478.x. [DOI] [PubMed] [Google Scholar]
  39. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  40. Subramani S., Mulligan R., Berg P. Expression of the mouse dihydrofolate reductase complementary deoxyribonucleic acid in simian virus 40 vectors. Mol Cell Biol. 1981 Sep;1(9):854–864. doi: 10.1128/mcb.1.9.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wigler M., Pellicer A., Silverstein S., Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. doi: 10.1016/0092-8674(78)90254-4. [DOI] [PubMed] [Google Scholar]
  43. Wijnen L. M., Monteba-van Heuvel M., Pearson P. L., Meera Khan P. Assignment of a gene for glutathione peroxidase (GPX1) to human chromosome 3. Cytogenet Cell Genet. 1978;22(1-6):232–235. doi: 10.1159/000130944. [DOI] [PubMed] [Google Scholar]
  44. Wright T. C., Orkin R. W., Destrempes M., Kurnit D. M. Increased adhesiveness of Down syndrome fetal fibroblasts in vitro. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2426–2430. doi: 10.1073/pnas.81.8.2426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yagi K. A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med. 1976 Apr;15(2):212–216. doi: 10.1016/0006-2944(76)90049-1. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES