Carbon Source-Dependent Regulation of the Schizosaccharomyces pombe pbh1 Gene

  • Kim, Su-Jung (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Cho, Nam-Chul (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Ryu, In-Wang (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Kim, Kyung-Hoon (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)
  • Published : 2006.12.31

Abstract

Pbh1, from the fission yeast Schizosaccharomyces pombe, is a baculoviral inhibitor of apoptosis (IAP) repeat (BIR) domain-containing protein. Its unique encoding gene was previously found to be regulated by nitric oxide and nitrogen starvation. In the current work, the Pbh1-lacZ fusion gene was used to elucidate the transcriptional regulation of the pbh1 gene under various carbon sources. When fermentable carbon sources, such as glucose (at a low concentration of 0.2 %), sucrose (2.0 %) and lactose (2.0 %), were the sole carbon source, the synthesis of $\beta$-galactosidase from the Pbh1-lacZ fusion gene was reasonably enhanced. However, the induction by these fermentable carbon sources was abolished in the Pap1-negative S. pombe cells, implying that this type of induction of the pbh1 gene is mediated by Pap1. Ethanol (2.0%), a nonfermentable carbon source, was also able to enhance the synthesis of $\beta$-galactosidase from the fusion gene in wild-type cells but not in Pap1-negative cells. The results indicate that the S. pombe pbh1 gene is up-regulated under metabolic oxidative stress in a Pap1-dependent manner.

Keywords

References

  1. Adams, R.R., S.P. Wheatley, A.M. Gouldsworthy, S.E. Kandels- Lewis, M. Carmena, C. Smythe, D.L. Gerloff, and W.C. Earnshaw. 2000. INCENP binds the aurora-related kinase AIRK2 and is required to target it to chromosomes, the central spindle and cleavage furrow. Curr. Biol. 10, 1075-1078 https://doi.org/10.1016/S0960-9822(00)00673-4
  2. 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
  3. Cho, N.C., H.J. Kang, H.Y. Lim, B.C. Kim, E.H. Park, and C.-J. Lim. 2006. Stress-dependent regulation of Pbh1, a BIR domain-containing protein, in the fission yeast. Can. J. Microbiol. (in press)
  4. Fujii, Y., T. Shimizu, T. Toda, M. Yanagida, and T. Hakoshima. 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
  5. Grant, C.M., F.H. Maciver, and I.W. Dawes. 1996 Stationaryphase induction of GLR1 expression is mediated by the yAP-1 transcriptional regulatory protein in the yeast Saccharomyces cerevisiae. Mol. Microbiol. 22, 739-746 https://doi.org/10.1046/j.1365-2958.1996.d01-1727.x
  6. 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
  7. Hiesinger, M., S. Roth, E. Meissner, and H.J. Schűller. 2001. Contribution of Cat8 and Sip4 to the transcriptional activation of yeast gluconeogenic genes by carbon sourceresponsive elements. Curr. Genet. 39, 68-76 https://doi.org/10.1007/s002940000182
  8. Johnston, M. and M. Carlson. 1992. Regulation of carbon and phosphate utilization. p. 193-281. In E.W. Jones, J.R. Pringle, and J.R. Broach (ed.), The molecular and cellular biology of the yeast Saccharomyces, vol. 2. Gene expression. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA
  9. Johnston, M. 1999. Feasting, fasting and fermenting. Glucose sensing in yeast and other cells. Trends Genet. 15, 29-33 https://doi.org/10.1016/S0168-9525(98)01637-0
  10. Lee, Y.J., S.S. Galoforo, C.M. Berns, J.C. Chen, B.H. Davis, J.E. Sim, P.M. Corry, and D.R. Spitz. 1998. Glucose deprivation- induced cytotoxicity and alterations in mitogenactivated 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
  11. Myers, A.M., A. Tzagoloff, D.M. Kinney, and C.J. Lusty. 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
  12. Nguyen, A.N., A. Lee, W. Place, and K. Shiozaki. 2000. Multistep phosphorelay proteins transmit oxidative stress signals to fission yeast stress-activated protein kinase. Mol. Biol. Cell 11, 1169-1181 https://doi.org/10.1091/mbc.11.4.1169
  13. Rajagopalan, S. and M.K. Balasubramanian. 1999. S. pombe Pbh1p: an inhibitor of apoptosis domain containing protein is essential for chromosome segregation. FEBS Lett. 460, 187-190 https://doi.org/10.1016/S0014-5793(99)01329-0
  14. Rajagopalan, S. and M.K. Balasubramanian. 2002. Schizosaccharomyces pombe Bir1p, a nuclear protein that localizes to kinetochores and the spindle midzone, is essential for chromosome condensation and spindle elongation during mitosis. Genetics 160, 445-456
  15. Reed, J.C. and J.R. Bischoff. 2000. BIRinging chromosomes through cell division-and survivin' the experience. Cell 102, 545-548 https://doi.org/10.1016/S0092-8674(00)00076-3
  16. Rolland, F., J. Winderickx, and J.M Thevelein. 2001. Glucosesensing mechanisms in eukaryotic cells. Trends Biochem. Sci. 13, 310-317
  17. Salvesen, G.S. and C.S. Duckett. 2002. IAP proteins: blocking the road to death's door. Nat. Rev. Mol. Cell Biol. 3, 401-410 https://doi.org/10.1038/nrm830
  18. Schuller, H.J. 2003. Transcriptional control of nonfermentative metabolism in the yeast Saccharomyces cerevisiae. Curr. Genet. 43, 139-160
  19. Song, J.J., J.G. Rhee, M. Suntharalingam, S.A. Walsh, D.R. Spitz, and Y.J. Lee. 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
  20. Thevelein, J.M., L. Cauwenberg, S. Colombo, J.H. De Winde, M. Donation, F. Dumortier, L. Kraakman, K. Lemaire, P. Ma, D. Nauwelaers, F. Rolland, A. Teunissen, P. Van Dijck, M. Versele, S. Wera, and J. Winderickx. 2000. Nutrient-induced signal transduction through the protein kinase A pathway and its role in the control of metabolism, stress resistance, and growth in yeast. Enzyme Microb. Technol. 26, 819-825 https://doi.org/10.1016/S0141-0229(00)00177-0
  21. Toone, W.M., S. Kuge, M. Samuels, B.A. Morgan, T. Toda, and N. Jones. 1998. Regulation of the fission yeast transcription factor Pap1 by oxidative stress: requirement for the nuclear export factor Crm1 (Exportin) and the stressactivated MAP kinase Sty1/ Spc1. Genes Dev. 12, 1391-1397 https://doi.org/10.1101/gad.12.10.1391
  22. Uren, A.G., E.J. Coulson, and D.L. Vaux. 1998. Conservation of baculovirus inhibitor of apoptosis repeat proteins (BIRPs) in viruses, nematodes, vertebrates and yeasts. Trends Biochem. Sci. 23, 159-162 https://doi.org/10.1016/S0968-0004(98)01198-0
  23. Walter, D., S. Wissing, F. Madeo, and B. Fahrenkrog. 2006. The inhibitor-of-apoptosis protein Bir1p protects against apoptosis in S. cerevisiae and is a substrate for the yeast homologue of Omi/HtrA2. J. Cell Sci. 119, 1843-1851 https://doi.org/10.1242/jcs.02902
  24. Walther, K. and H.J. Schuller. 2001. Adr1 and Cat8 synergistically activate the glucose-regulated alcohol dehydrogenase gene ADH2 of the yeast Saccharomyces cerevisiae. Microbiology 147, 2037-2044 https://doi.org/10.1099/00221287-147-8-2037
  25. Wiatrowski, H.A. and M. Carlson. 2003. Yap1 accumulates in the nucleus in response to carbon stress in Saccharomyces cerevisiae. Euk. Cell 2, 19-26 https://doi.org/10.1128/EC.2.1.19-26.2003