Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Mar;15(3):1382–1388. doi: 10.1128/mcb.15.3.1382

Mutations in PMR1 suppress oxidative damage in yeast cells lacking superoxide dismutase.

P J Lapinskas 1, K W Cunningham 1, X F Liu 1, G R Fink 1, V C Culotta 1
PMCID: PMC230362  PMID: 7862131

Abstract

Mutants of Saccharomyces cerevisiae lacking a functional SOD1 gene encoding Cu/Zn superoxide dismutase (SOD) are sensitive to atmospheric levels of oxygen and are auxotrophic for lysine and methionine when grown in air. We have previously shown that these defects of SOD-deficient yeast cells can be overcome through mutations in either the BSD1 or BSD2 (bypass SOD defects) gene. In this study, the wild-type allele of BSD1 was cloned by functional complementation and was physically mapped to the left arm of chromosome VII. BSD1 is identical to PMR1, encoding a member of the P-type ATPase family that localizes to the Golgi apparatus. PMR1 is thought to function in calcium metabolism, and we provide evidence that PMR1 also participates in the homeostasis of manganese ions. Cells lacking a functional PMR1 gene accumulate elevated levels of intracellular manganese and are also extremely sensitive to manganese ion toxicity. We demonstrate that mutations in PMR1 bypass SOD deficiency through a mechanism that depends on extracellular manganese. Collectively, these findings indicate that oxidative damage in a eukaryotic cell can be prevented through alterations in manganese homeostasis.

Full Text

The Full Text of this article is available as a PDF (238.3 KB).

Selected References

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

  1. Ames B. N., Shigenaga M. K., Hagen T. M. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7915–7922. doi: 10.1073/pnas.90.17.7915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Antebi A., Fink G. R. The yeast Ca(2+)-ATPase homologue, PMR1, is required for normal Golgi function and localizes in a novel Golgi-like distribution. Mol Biol Cell. 1992 Jun;3(6):633–654. doi: 10.1091/mbc.3.6.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Archibald F. S., Fridovich I. Manganese and defenses against oxygen toxicity in Lactobacillus plantarum. J Bacteriol. 1981 Jan;145(1):442–451. doi: 10.1128/jb.145.1.442-451.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Archibald F. S., Fridovich I. The scavenging of superoxide radical by manganous complexes: in vitro. Arch Biochem Biophys. 1982 Apr 1;214(2):452–463. doi: 10.1016/0003-9861(82)90049-2. [DOI] [PubMed] [Google Scholar]
  5. Becker D. M., Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–187. doi: 10.1016/0076-6879(91)94015-5. [DOI] [PubMed] [Google Scholar]
  6. Bermingham-McDonogh O., Gralla E. B., Valentine J. S. The copper, zinc-superoxide dismutase gene of Saccharomyces cerevisiae: cloning, sequencing, and biological activity. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4789–4793. doi: 10.1073/pnas.85.13.4789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Biliński T., Krawiec Z., Liczmański A., Litwińska J. Is hydroxyl radical generated by the Fenton reaction in vivo? Biochem Biophys Res Commun. 1985 Jul 31;130(2):533–539. doi: 10.1016/0006-291x(85)90449-8. [DOI] [PubMed] [Google Scholar]
  8. Bostek C. C. Oxygen toxicity: an introduction. AANA J. 1989 Jun;57(3):231–237. [PubMed] [Google Scholar]
  9. Breimer L. H. Molecular mechanisms of oxygen radical carcinogenesis and mutagenesis: the role of DNA base damage. Mol Carcinog. 1990;3(4):188–197. doi: 10.1002/mc.2940030405. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Cerutti P., Larsson R., Krupitza G., Muehlematter D., Crawford D., Amstad P. Pathophysiological mechanisms of active oxygen. Mutat Res. 1989 Sep;214(1):81–88. doi: 10.1016/0027-5107(89)90200-5. [DOI] [PubMed] [Google Scholar]
  12. Chang E. C., Crawford B. F., Hong Z., Bilinski T., Kosman D. J. Genetic and biochemical characterization of Cu,Zn superoxide dismutase mutants in Saccharomyces cerevisiae. J Biol Chem. 1991 Mar 5;266(7):4417–4424. [PubMed] [Google Scholar]
  13. Chang E. C., Kosman D. J. Intracellular Mn (II)-associated superoxide scavenging activity protects Cu,Zn superoxide dismutase-deficient Saccharomyces cerevisiae against dioxygen stress. J Biol Chem. 1989 Jul 25;264(21):12172–12178. [PubMed] [Google Scholar]
  14. Chang E. C., Kosman D. J. O2-dependent methionine auxotrophy in Cu,Zn superoxide dismutase-deficient mutants of Saccharomyces cerevisiae. J Bacteriol. 1990 Apr;172(4):1840–1845. doi: 10.1128/jb.172.4.1840-1845.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Chiesi M., Inesi G. Mg2+ and Mn2+ modulation of Ca2+ transport and ATPase activity in sarcoplasmic reticulum vesicles. Arch Biochem Biophys. 1981 May;208(2):586–592. doi: 10.1016/0003-9861(81)90547-6. [DOI] [PubMed] [Google Scholar]
  16. Cross C. E., Halliwell B., Borish E. T., Pryor W. A., Ames B. N., Saul R. L., McCord J. M., Harman D. Oxygen radicals and human disease. Ann Intern Med. 1987 Oct;107(4):526–545. doi: 10.7326/0003-4819-107-4-526. [DOI] [PubMed] [Google Scholar]
  17. Cunningham K. W., Fink G. R. Calcineurin-dependent growth control in Saccharomyces cerevisiae mutants lacking PMC1, a homolog of plasma membrane Ca2+ ATPases. J Cell Biol. 1994 Feb;124(3):351–363. doi: 10.1083/jcb.124.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Dancis A., Klausner R. D., Hinnebusch A. G., Barriocanal J. G. Genetic evidence that ferric reductase is required for iron uptake in Saccharomyces cerevisiae. Mol Cell Biol. 1990 May;10(5):2294–2301. doi: 10.1128/mcb.10.5.2294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Davies K. J. Protein damage and degradation by oxygen radicals. I. general aspects. J Biol Chem. 1987 Jul 15;262(20):9895–9901. [PubMed] [Google Scholar]
  20. 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]
  21. Dunn T., Gable K., Beeler T. Regulation of cellular Ca2+ by yeast vacuoles. J Biol Chem. 1994 Mar 11;269(10):7273–7278. [PubMed] [Google Scholar]
  22. Emery H. S., Schild D., Kellogg D. E., Mortimer R. K. Sequence of RAD54, a Saccharomyces cerevisiae gene involved in recombination and repair. Gene. 1991 Jul 31;104(1):103–106. doi: 10.1016/0378-1119(91)90473-o. [DOI] [PubMed] [Google Scholar]
  23. Evans H. E., Rosenfeld W., Jhaveri R., Concepcion L., Zabaleta I. Oxidant-mediated lung disease in newborn infants. J Free Radic Biol Med. 1986;2(5-6):369–372. doi: 10.1016/s0748-5514(86)80038-1. [DOI] [PubMed] [Google Scholar]
  24. Farr S. B., D'Ari R., Touati D. Oxygen-dependent mutagenesis in Escherichia coli lacking superoxide dismutase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8268–8272. doi: 10.1073/pnas.83.21.8268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Fridovich I. Superoxide radical: an endogenous toxicant. Annu Rev Pharmacol Toxicol. 1983;23:239–257. doi: 10.1146/annurev.pa.23.040183.001323. [DOI] [PubMed] [Google Scholar]
  26. Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
  27. Fürst P., Hu S., Hackett R., Hamer D. Copper activates metallothionein gene transcription by altering the conformation of a specific DNA binding protein. Cell. 1988 Nov 18;55(4):705–717. doi: 10.1016/0092-8674(88)90229-2. [DOI] [PubMed] [Google Scholar]
  28. Gomes da Costa A., Madeira V. M. Magnesium and manganese ions modulate Ca2+ uptake and its energetic coupling in sarcoplasmic reticulum. Arch Biochem Biophys. 1986 Aug 15;249(1):199–206. doi: 10.1016/0003-9861(86)90575-8. [DOI] [PubMed] [Google Scholar]
  29. Gralla E. B., Kosman D. J. Molecular genetics of superoxide dismutases in yeasts and related fungi. Adv Genet. 1992;30:251–319. doi: 10.1016/s0065-2660(08)60322-3. [DOI] [PubMed] [Google Scholar]
  30. Gralla E. B., Valentine J. S. Null mutants of Saccharomyces cerevisiae Cu,Zn superoxide dismutase: characterization and spontaneous mutation rates. J Bacteriol. 1991 Sep;173(18):5918–5920. doi: 10.1128/jb.173.18.5918-5920.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Gunteski-Hamblin A. M., Clarke D. M., Shull G. E. Molecular cloning and tissue distribution of alternatively spliced mRNAs encoding possible mammalian homologues of the yeast secretory pathway calcium pump. Biochemistry. 1992 Aug 25;31(33):7600–7608. doi: 10.1021/bi00148a023. [DOI] [PubMed] [Google Scholar]
  32. Halliwell B., Gutteridge J. M. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984 Apr 1;219(1):1–14. doi: 10.1042/bj2190001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Halliwell B. Oxidants and human disease: some new concepts. FASEB J. 1987 Nov;1(5):358–364. [PubMed] [Google Scholar]
  34. Harman D. Free radical theory of aging. Mutat Res. 1992 Sep;275(3-6):257–266. doi: 10.1016/0921-8734(92)90030-s. [DOI] [PubMed] [Google Scholar]
  35. Haselbeck A., Schekman R. Interorganelle transfer and glycosylation of yeast invertase in vitro. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2017–2021. doi: 10.1073/pnas.83.7.2017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Imlay J. A., Linn S. DNA damage and oxygen radical toxicity. Science. 1988 Jun 3;240(4857):1302–1309. doi: 10.1126/science.3287616. [DOI] [PubMed] [Google Scholar]
  37. Jamieson D. Oxygen toxicity and reactive oxygen metabolites in mammals. Free Radic Biol Med. 1989;7(1):87–108. doi: 10.1016/0891-5849(89)90103-2. [DOI] [PubMed] [Google Scholar]
  38. Kaufman R. J., Swaroop M., Murtha-Riel P. Depletion of manganese within the secretory pathway inhibits O-linked glycosylation in mammalian cells. Biochemistry. 1994 Aug 23;33(33):9813–9819. doi: 10.1021/bi00199a001. [DOI] [PubMed] [Google Scholar]
  39. Koppenol W. H., Levine F., Hatmaker T. L., Epp J., Rush J. D. Catalysis of superoxide dismutation by manganese aminopolycarboxylate complexes. Arch Biochem Biophys. 1986 Dec;251(2):594–599. doi: 10.1016/0003-9861(86)90368-1. [DOI] [PubMed] [Google Scholar]
  40. Lin C. M., Kosman D. J. Copper uptake in wild type and copper metallothionein-deficient Saccharomyces cerevisiae. Kinetics and mechanism. J Biol Chem. 1990 Jun 5;265(16):9194–9200. [PubMed] [Google Scholar]
  41. Link A. J., Olson M. V. Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution. Genetics. 1991 Apr;127(4):681–698. doi: 10.1093/genetics/127.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Liu X. F., Culotta V. C. The requirement for yeast superoxide dismutase is bypassed through mutations in BSD2, a novel metal homeostasis gene. Mol Cell Biol. 1994 Nov;14(11):7037–7045. doi: 10.1128/mcb.14.11.7037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Liu X. F., Elashvili I., Gralla E. B., Valentine J. S., Lapinskas P., Culotta V. C. Yeast lacking superoxide dismutase. Isolation of genetic suppressors. J Biol Chem. 1992 Sep 15;267(26):18298–18302. [PubMed] [Google Scholar]
  45. Lumsden J., Hall D. O. Chloroplast manganese and superoxide. Biochem Biophys Res Commun. 1975 May 19;64(2):595–602. doi: 10.1016/0006-291x(75)90363-0. [DOI] [PubMed] [Google Scholar]
  46. Nakajima T., Ballou C. E. Yeast manno-protein biosynthesis: solubilization and selective assay of four mannosyltransferases. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3912–3916. doi: 10.1073/pnas.72.10.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Nakazono K., Watanabe N., Matsuno K., Sasaki J., Sato T., Inoue M. Does superoxide underlie the pathogenesis of hypertension? Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10045–10048. doi: 10.1073/pnas.88.22.10045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Nasmyth K. A., Reed S. I. Isolation of genes by complementation in yeast: molecular cloning of a cell-cycle gene. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2119–2123. doi: 10.1073/pnas.77.4.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Parodi A. J. Biosynthesis of yeast mannoproteins. Synthesis of mannan outer chain and of dolichol derivatives. J Biol Chem. 1979 Sep 10;254(17):8343–8352. [PubMed] [Google Scholar]
  50. Phillips J. P., Campbell S. D., Michaud D., Charbonneau M., Hilliker A. J. Null mutation of copper/zinc superoxide dismutase in Drosophila confers hypersensitivity to paraquat and reduced longevity. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2761–2765. doi: 10.1073/pnas.86.8.2761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Riles L., Dutchik J. E., Baktha A., McCauley B. K., Thayer E. C., Leckie M. P., Braden V. V., Depke J. E., Olson M. V. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs. Genetics. 1993 May;134(1):81–150. doi: 10.1093/genetics/134.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]
  53. Rudolph H. K., Antebi A., Fink G. R., Buckley C. M., Dorman T. E., LeVitre J., Davidow L. S., Mao J. I., Moir D. T. The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell. 1989 Jul 14;58(1):133–145. doi: 10.1016/0092-8674(89)90410-8. [DOI] [PubMed] [Google Scholar]
  54. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tamai K. T., Gralla E. B., Ellerby L. M., Valentine J. S., Thiele D. J. Yeast and mammalian metallothioneins functionally substitute for yeast copper-zinc superoxide dismutase. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8013–8017. doi: 10.1073/pnas.90.17.8013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Thiele D. J. ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene. Mol Cell Biol. 1988 Jul;8(7):2745–2752. doi: 10.1128/mcb.8.7.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Trush M. A., Kensler T. W. An overview of the relationship between oxidative stress and chemical carcinogenesis. Free Radic Biol Med. 1991;10(3-4):201–209. doi: 10.1016/0891-5849(91)90077-g. [DOI] [PubMed] [Google Scholar]
  58. Trush M. A., Mimnaugh E. G., Gram T. E. Activation of pharmacologic agents to radical intermediates. Implications for the role of free radicals in drug action and toxicity. Biochem Pharmacol. 1982 Nov 1;31(21):3335–3346. doi: 10.1016/0006-2952(82)90609-8. [DOI] [PubMed] [Google Scholar]
  59. Welch J., Fogel S., Buchman C., Karin M. The CUP2 gene product regulates the expression of the CUP1 gene, coding for yeast metallothionein. EMBO J. 1989 Jan;8(1):255–260. doi: 10.1002/j.1460-2075.1989.tb03371.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. van Loon A. P., Pesold-Hurt B., Schatz G. A yeast mutant lacking mitochondrial manganese-superoxide dismutase is hypersensitive to oxygen. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3820–3824. doi: 10.1073/pnas.83.11.3820. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES