Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Characterization and Regulation of the Gene Encoding Monothiol Glutaredoxin 3 in the Fission Yeast Schizosaccharomyces pombe

  • Moon, Jeong-Su (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Lim, Hye-Won (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Park, Eun-Hee (College of Pharmacy, Sookmyung Women's University) ;
  • Lim, Chang-Jin (Division of Life Sciences, College of Natural Sciences, Kangwon National University)
  • Received : 2005.02.19
  • Accepted : 2005.04.13
  • Published : 2005.08.31
⟨ Previous Next ⟩

Abstract

Glutaredoxins (Grxs) are thioloxidoreductases which are required for maintaining thiol/disulfide equilibrium in living cells. The Grx3 gene, which encodes one of the three monothiol Grxs in the fission yeast Schizosaccharomyces pombe, was characterized, and its transcriptional regulation studied. Genomic DNA encoding Grx3 was isolated by PCR, and a plasmid pTT3 carrying this DNA was produced. The DNA sequence has 1,267 bp, which would encode a monothiol Grx of 166 amino acids with a molecular mass of 18.3 kDa. The putative protein has 27% homology with Grx5, and contains many hydrophobic amino acid residues in its N-terminal region. S. pombe cells harboring pTT3 had increased Grx activity and enhanced survival on minimal medium plates containing aluminum (5 mM), BSO (0.05 mM), menadione (0.01 mM) or cadmium (0.2 mM). The 568 bp upstream region of Grx3 was fused into the promoterless b-galactosidase gene of the shuttle vector YEp367R to generate fusion plasmid pMJS10. Potassium chloride (KCl) and metals including aluminum and cadmium enhanced the synthesis of ${\beta}$-galactosidase from the fusion gene. The synthesis of ${\beta}$-galactosidase was also enhanced, in a Pap1-dependent manner, by fermentable carbon sources such as glucose (at low concentrations) and sucrose, but not by non-fermentable carbon sources such as ethanol and acetate. Grx3 mRNA increased in response to treatment with BSO. These observations indicate that S. pombe Grx3 is involved in the response to stress, and is regulated by stress.

Keywords

Acknowledgement

Supported by : Korea Research Foundation

References

  1. Aslund, F., Ehn, B., Miranda-Vizuete, A., Pueyo, C., and Holmgren, A. (1994) Two additional glutaredoxins exist in Escherichia coli: glutaredoxin 3 is a hydrogen donor for ribonucleotide reductase in a thioredoxin/glutaredoxin 1 double mutant. Proc. Natl. Acad. Sci. USA 91, 9813-9817
  2. Bandyopadhyay, S., Starke, D. W., Mieyal, J. J., and Gronostajski, R. M. (1998) Thioltransferase (glutaredoxin) reactivates the DNA-binding activity of oxidation-inactivated nuclear factor I. J. Biol. Chem. 273, 392-397 https://doi.org/10.1074/jbc.273.1.392
  3. Belli, G., Polaina, J., Tamarit, J., de la Torre, M. A., Rodríguez- Manzaneque, M. T., et al. (2002) Structure-function analysis of yeast Grx5 monothiol glutaredoxin defines essential amino acids for the function of the protein. J. Biol. Chem. 277, 37590-37596 https://doi.org/10.1074/jbc.M201688200
  4. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248- 254 https://doi.org/10.1016/0003-2697(76)90527-3
  5. Cho, Y. W., Park, E. H., and Lim, C. J. (2000) Glutathione Stransferase activities of S-type and L-type thioltransferases from Arabidopsis thaliana. J. Biochem. Mol. Biol. 33, 179- 183
  6. Daily, D., Vlamis-Gardikas, A., Offen, D., Mittelman, L., Melamed, E., et al. (2001) Glutaredoxin protects cerebellar granule neurons from dopamine-induced apoptosis by activating NF-kappa B via Ref-1. J. Biol. Chem. 276, 1335-1344 https://doi.org/10.1074/jbc.M008121200
  7. Degols, G. and Russell, P. (1997) Discrete roles of the Spc1 kinase and the Atf1 transcription factor in the UV response of Schizosaccharomyces pombe. Mol. Cell. Biol. 17, 3356-3363
  8. Fujii, Y., Shimizu, T., Toba, T., Yanagida, M., and Hakoshima, T. (2000) Structural basis for the diversity of DNA recognition by bZIP transcription factors. Nat. Struct. Biol. 7, 889-893 https://doi.org/10.1038/82822
  9. Gan, Z. R. and Wells, W. W. (1986) Purification and properties of thioltransferase. J. Biol. Chem. 261, 996-1001
  10. Gravina, S. A. and Mieyal, J. J. (1993) Thioltransferase is a specific glutathionyl mixed-disulfide oxidoreductase. Biochemistry 32, 3368-3376 https://doi.org/10.1021/bi00064a021
  11. Guarente, L. (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 101, 181-191 https://doi.org/10.1016/0076-6879(83)01013-7
  12. Gupta, S., Campbell, D., Derijard, B., and Davis, R. J. (1995) Transcription factor ATF2 regulation by the JNK signal transduction pathway. Science 267, 389-393 https://doi.org/10.1126/science.7824938
  13. Holmgren, A. (1976) Hydrogen donor system for Escherichia coli ribonucleoside-diphosphate reductase dependent upon glutathione. Proc. Natl. Acad. Sci. USA 73, 2275-2279
  14. Holmgren, A. (1979) Glutathione-dependent synthesis of deoxyribonucleotides. Characterization of the enzymatic mechanism of Escherichia coli glutaredoxin. J. Biol. Chem. 254, 3672-3678
  15. Holmgren, A. and Barcena, J. A. (2002) Glutaredoxins catalyze the reduction of glutathione by dihydrolipoamide with high efficiency. Biochem. Biophys. Res. Commun. 295, 1046-1051 https://doi.org/10.1016/S0006-291X(02)00771-4
  16. Isakov, N., Witte, S., and Altman, A. (2000) PICOT-HD: a highly conserved protein domain that is often associated with thioredoxin and glutaredoxin modules. Trends Biochem. Sci. 25, 537-539 https://doi.org/10.1016/S0968-0004(00)01685-6
  17. Kato, T., Okazaki, K., Murakami, H., Stettler, S., Fantes, P., et al. (1996) Stress signal, mediated by a Hog1-like MAP kinase, controls sexual development in fission yeast. FEBS Lett. 378, 207-212 https://doi.org/10.1016/0014-5793(95)01442-X
  18. Kenchappa, R. S., Diwakar, L., Boyd, M. R., and Ravindranath, V. (2002) Thioltransferase (Glutaredoxin) mediates recovery of motor neurons from excitotoxic mitochondrial injury. J. Neurosci. 22, 8402-8410
  19. Kim, H. G., Park, E. H., and Lim, C. J. (1999) A second thioltransferase of Schizosaccharomyces pombe contains glutathione S-transferase activity. J. Biochem. Mol. Biol. 32, 535-540
  20. Kim, H.-G., Kim, B.-C., Park, E.-H., and Lim, C.-J. (2005a) Stress-dependent regulation of a monothiol glutaredoxin gene from Schizosaccharomyces pombe. Can. J. Microbiol. (in press)
  21. Kim, H.-G., Kim, J.-H., Kim, B.-C., Park, E.-H., and Lim, C.-J. (2005b) Carbon source-dependent regulation of a second gene encoding glutaredoxin from the fission yeast Schizosaccharomyces pombe. Mol. Biol. Rep. (in press)
  22. Lee, Y. J., Galoforo, S. S., Berns, C. M., Chen, J. C., Davis, B. H., et al. (1998) Glucose deprivation-induced cytotoxicity and alterations in mitogen-activated protein kinase activation are mediated by oxidative stress in multidrug-resistant human breast carcinoma Cells. J. Biol. Chem. 273, 5294-5299 https://doi.org/10.1074/jbc.273.9.5294
  23. Lee, D. H., Kang, S.-G.., Suh, S.-G., and Kang, J. K. (2003) Purification and characterization of a ${\beta}$-galactosidase from peach (Prunus persica). Mol. Cells 15, 68-74
  24. Lillig, C. H., Prior, A., Schween, J. D., Aslund, F., and Holmgren, A. (1999) New thioredoxins and glutaredoxins as electron donors of 3′-phosphoadenylylsulfate reductase. J. Biol. Chem. 274, 7695-7698 https://doi.org/10.1074/jbc.274.12.7695
  25. Livingstone, C., Patel, G., and Jones, N. (1995) ATF-2 contains a phosphorylation-dependent transcriptional activation domain. EMBO J. 14, 1785–1797
  26. Luikenhuis, S., Perrone, G., Dawes, I. W., and Grant, C. M. (1998) The yeast Saccharomyces cerevisiae contains two glutaredoxin genes that are required for protection against reactive oxygen species. Mol. Biol. Cell. 9, 1081-1091
  27. Madrid, M., Soto, T., Franco, A., Paredes, V., Hidalgo, E., et al. (2004) A cooperative role for Atf1 and Pap1 in the detoxification of the oxidative stress induced by glucose deprivation in Schizosaccharomyces pombe. J. Biol. Chem. 279, 41594- 41602 https://doi.org/10.1074/jbc.M405509200
  28. Millar, J., Buck, V., and Wilkinson, M. (1995) Pyp1 and Pyp2 PTPases dephosphorylate an osmosensing MAP kinase controlling cell size at division in fission yeast. Genes Dev. 9, 2117-2130 https://doi.org/10.1101/gad.9.17.2117
  29. Moradas-Ferreira, P., Costa, V., Piper, P., and Mager, W. (1996) The molecular defences against reactive oxygen species in yeast. Microbiology 19, 651-658
  30. Murata, H., Ihara, Y., Nakamura, H., Yodoi, J., Sumikawa, K., et al. (2003) Glutaredoxin exerts an antiapoptotic effect by regulating the redox state of Akt. J. Biol. Chem. 278, 50226-50233 https://doi.org/10.1074/jbc.M310171200
  31. Myers, A. M., Tzagoloff, A., Kinney, D. M., and Lusty, C. J. (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene 45, 299-310 https://doi.org/10.1016/0378-1119(86)90028-4
  32. Nguyen, A. N., Lee, A., Place, W., and Shiozaki, K. (2000) Multistep phosphorelay proteins transmit oxidative stress signals to the fission yeast stress-activated protein kinase. Mol. Biol. Cell 11, 1169-1181
  33. Porras, P., Pedrajas, J. R., Martínez-Galisteo, E., Padilla, C. A., Johansson, C., et al. (2002) Glutaredoxins catalyze the reduction of glutathione by dihydrolipoamide with high efficiency. Biochem. Biophys. Res. Commun. 295, 1046-1051 https://doi.org/10.1016/S0006-291X(02)00771-4
  34. Raingeaud, J., Whitmarsh, A. J., Barrett, T., Derijard, B., and Davis, R. J. (1996) MKK3- and MKK6-regulated gene expression is mediated by the p38 mitogen- activated protein kinase signal transduction pathway. Mol. Cell. Biol. 16, 1247-1255
  35. Rodriguez-Manzaneque, M. T., Ros, J., Cabiscol, E., Sorribas, A., and Herrero, E. (1999) Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. Mol. Cell. Biol. 19, 8180-8190
  36. Rodríguez-Manzaneque, M. T., Tamarit, J., Bellí, G., Ros, J., and Herrero, E. (2002) Grx5 is a mitochondrial glutaredoxin required for the activity of iron/sulfur enzymes. Mol. Biol. Cell 13, 1109-1121 https://doi.org/10.1091/mbc.01-10-0517
  37. Rolland, F., Winderickx, J., and Thevelein, J. M. (2001) Glucose- sensing mechanisms in eukaryotic cells. Trends Biochem. Sci. 26, 310–317 https://doi.org/10.1016/S0968-0004(01)01805-9
  38. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
  39. Shiozaki, K. and Russell, P. (1995) Cell-cycle control linked to extracellular environment by MAP kinase pathway in fission yeast. Nature 378, 739-743 https://doi.org/10.1038/378739a0
  40. Shiozaki, K., Shiozaki, M., and Russell, P. (1997) Mcs4 mitotic catastrophe suppressor regulates the fission yeast cell cycle through the Wik1-Wis1-Spc1 kinase cascade. Mol. Biol. Cell 8, 409–419
  41. Song, J. J., Rhee, J. G., Suntharalingam, M., Walsh, S. A., Spitz, D. R., et al. (2002) Role of glutaredoxin in metabolic oxidative stress. Glutaredoxin as a sensor of oxidative stress mediated by $H_2O_2$. J. Biol. Chem. 277, 46566-46575 https://doi.org/10.1074/jbc.M206826200
  42. Spitz, D. R., Sim, J. E., Ridnour, L. A., Galoforo, S. S., and Lee, Y. J. (2000) Glucose deprivation-induced oxidative stress in human tumor cells: a fundamental defect in metabolism? Ann. N. Y. Acad. Sci. 899, 349–362
  43. Stettler, S., Warbrick, E., Prochnik, S., Mackie, S., and Fantes, P. (1996) The wis1 signal transduction pathway is required for expression of cAMP-repressed genes in fission yeast. J. Cell Sci. 109, 1927–1935
  44. Toda, T., Shimanuki, M., and Yanagida, M. (1991) Fission yeast genes that confer resistance to staurosporine encode an AP-1- like transcription factor and a protein kinase related to the mammalian ERK1/MAP2 and budding yeast FUS3 and KSS1 kinases. Genes Dev. 5, 60-73 https://doi.org/10.1101/gad.5.1.60
  45. Toone, W. M., Kuge, S., Samuels, M., Morgan, B. A., Toda, T., et al. (1998) Regulation of the fission yeast transcription factor Pap1 by oxidative stress: requirement for the nuclear export factor Crm1 (exportin) and the stress-activated MAP kinase Sty1/Spc1. Genes Dev. 12, 23042-23049
  46. Van Dam, H., Wilhelm, D., Herr, I., Steffen, A., Herrlich, P., et al. (1995) ATF-2 is preferentially activated by stressactivated protein kinases to mediate c-jun induction in response to genotoxic agents. EMBO J. 14, 1798–1811
  47. Vincent, O. and Carlson, M. (1998) Sip4, a Snf1 kinasedependent transcriptional activator, binds to the carbon sourceresponsive element of gluconeogenic genes. EMBO J. 17, 7002-7008 https://doi.org/10.1093/emboj/17.23.7002
  48. Walther, K. and Schuller, H. J. (2001) Adr1 and Cat8 synergistically activate the glucose-regulated alcohol dehydrogenase gene ADH2 of the yeast Saccharomyces cerevisiae. Microbiology 147, 2037-2044
  49. Wells, W. W., Xu, D. P., Yang, Y. F., and Rocque, P. A. (1990) Mammalian thioltransferase (glutaredoxin) and protein disulfide isomerase have dehydroascorbate reductase activity. J. Biol. Chem. 265, 15361-15364
  50. Witte, S., Villalba, M., Bi, K., Liu, Y., Isakov, N., et al. (2000) Inhibition of the c-Jun N-terminal kinase/AP-1 and NF-${\kappa}B$ pathways by PICOT, a novel protein kinase C-interacting protein with a thioredoxin homology domain. J. Biol. Chem. 275, 1902-1909 https://doi.org/10.1074/jbc.275.3.1902