HSF1-mediated oxidative stress response to menadione in Saccharomyces cerevisiae KNU5377Y3 by using proteomic approach


The hat shock transcription factor HSF1 inthe yeast Saccharomyces cerevisiae regulates a wide range of genes and functions in diverse cellular reactions. To investigate the physiological response of HSF1 inthe presence of menadione (MD) in S. cerevisiae KNU 5377Y3, wild-type (k3wt) and isogenic hsf1 mutant (k3h1) cells were introduced. HSF1 was induced when k3wt cells were exposed to the superoxide-generating agent MD and k3h1 cells were hypersensitive to MD. Under MD stress, k3h1 cells down-regulated the expression of metabolic enzymes (Hxk, Fba1, Pgk1, Eno2, and Adh1), antioxidant enzymes (Trx2 and porin), and molecular chaperones and their cofactors (Hsp104, Ssb1, Hsp60, Hsp42, Hsp26, Hsp12, Cpr1, and Sti1). In addition, k3h1 cells increased cellular hydroperoxide levels and protein carbonylation under MD stress as compared to k3wt cells. However, there was a moderate difference in the wild-type (b3wt) and mutant (b3h1) cells derived from S. cerevisiae S288Cunder the same conditions. Thus, these results show that HSF1 is an important component of the stress response system, acting as an activator of cell rescue genes in S. cerevisiae KNU5377Y3, and its expression protects the cells from MD-induced oxidative damage by maintaining redox homeostasis and proteostasis in the presence of MD.

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Kim, I. , Kim, H. , Kim, Y. , Jin, I. and Yoon, H. (2013) HSF1-mediated oxidative stress response to menadione in Saccharomyces cerevisiae KNU5377Y3 by using proteomic approach. Advances in Bioscience and Biotechnology, 4, 44-54. doi: 10.4236/abb.2013.41007.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Yan, L.J., Christians, E.S., Liu, L., Xiao, X., Sohal, R.S. and Benjamin, I.J. (2002) Mouse heat shock transcription factor 1 deficiency alters cardiac redox homeostasis and increases mitochondrial oxidative damage. EMBO Journal, 21, 5164-5172. doi:10.1093/emboj/cdf528
[2] Rodrigues-Pousada, C.A., Nevitt, T., Menezes, R., Azevedo, D., Pereira, J. and Amaral, C. (2004) Yeast activator proteins and stress response: An overview. FEBS Letters, 567, 80-85. doi:10.1016/j.febslet.2004.03.119
[3] Finkel, T. and Holbrook, N.J. (2000) Oxidants, oxidative stress and the biology of ageing. Nature, 408, 239-247. doi:10.1038/35041687
[4] Lee, S., Carlson, T., Christian, N., Lea, K., Kedzie, J., Reilly, J.P. and Bonner, J.J. (2000) The yeast heat shock transcription factor changes conformation in response to superoxide and temperature. Molecular Biology of the Cell, 11, 1753-1764.
[5] Yamamoto, A., Ueda, J., Yamamoto, N., Hashikawa, N. and Sakurai, H. (2007) Role of heat shock transcription factor in Saccharomyces cerevisiae oxidative stress response. Eukaryotic Cells, 6, 1373-1379. doi:10.1128/EC.00098-07
[6] Wu, C. (1995) Heat shock transcription factors: Structure and regulation. Annual Review of Cell and Developmental Biology, 11, 441-469. doi:10.1146/annurev.cb.11.110195.002301
[7] Hahn, J.S., Neef, D.W. and Thiele, D.J. (2006) A stress regulatory network for co-ordinated activation of proteasome expression mediated by yeast heat shock transcription factor. Molecular Microbiology, 60, 240-251. doi:10.1111/j.1365-2958.2006.05097.x
[8] Liu, X.D. and Thiele, D.J. (1996) Oxidative stress induced heat shock factor phosphorylation and HSF-dependent activation of yeast metallothionein gene transcription. Genes and Development, 10, 592-603. doi:10.1101/gad.10.5.592
[9] Takemori, Y., Sakaguchi, A., Matsuda, S., Mizukami, Y. and Sakurai, H. (2006) Stress-induced transcription of the endoplasmic reticulum oxidoreductin gene ERO1 in the yeast Saccharomyces cerevisiae. Molecular Genetics and Genomics, 275, 89-96. doi:10.1007/s00438-005-0065-9
[10] Kim, I.S., Jin, I. and Yoon, H.S. (2011) Decarbonylated cyclophilin A Cpr1 protein protects Saccharomyces cerevisiae KNU5377Y when exposed to stress induced by menadione. Cell Stress and Chaperones, 16, 1-14. doi:10.1007/s12192-010-0215-9
[11] Kim, I.S., Yun, H.S., Choi, H.J., Sohn, H.Y., Yu, C.B., Kim, J.K. and Jin, I.N. (2006) Construction of hsf1 knockout-mutant of a thermotolerant yeast strain Saccharomyces cerevisiae KNU5377. Korean Journal of Life Science, 16, 454-458.
[12] Kim, I.S., Moon, H.Y., Yun, H.S. and Jin, I. (2006) Heat shock causes oxidative stress and induces a variety of cell rescue proteins in Saccharomyces cerevisiae KNU5377. Journal of Microbiology, 44, 492-501.
[13] Kim. I., Yun, H. and Jin, I. (2007) Comparative proteomic analyses of the yeast Saccharomyces cerevisiae KNU5377 strain against menadione-induced oxidative stress. Journal of Microbiology and Biotechnology, 17, 207-217.
[14] Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. doi:10.1038/227680a0
[15] Ramagli, L. and Rodriguez, L. (1985) Quantitation of microgram amounts of protein in two-dimensional polyacrtlamide gel electrophoresis sample buffer. Electrophoresis, 6, 559-563. doi:10.1002/elps.1150061109
[16] Kim, I.S., Yun, H.S., Park, I.S., Sohn, H.Y., Iwahashi, H. and Jin, I.N. (2006) A knockout strain of CPR1 induced during fermentation of Saccharomyces cerevisiae KNU5377 is susceptible to various types of stress. Journal of Bioscience and Bioengineering, 102, 288-296. doi:10.1263/jbb.102.288
[17] Hahn, J.S. and Thiele, D.J. (2004) Activation of the Saccharomyces cerevisiae heat shock transcription factor under glucose starvation conditions by Snf1 protein kinase. The Journal of Biological Chemistry, 279, 5169-5176. doi:10.1074/jbc.M311005200
[18] Zhong, M., Orosz, A. and Wu, C. (1998) Direct sensing of heat and oxidation by Drosophila heat shock transcription factor. Molecualr Cell, 2, 101-108. doi:10.1016/S1097-2765(00)80118-5
[19] Ahn, S.G. and Thiele, D.J. (2003) Redox regulation of mammalian heat shock factor 1 is essential for Hsp gene activation and protection from stress. Genes and Development, 17, 516-528. doi:10.1101/gad.1044503
[20] Christians, E.S., Yan, L.J. and Benjamin, I.J. (2002) Heat shock factor 1 and heat shock proteins: Critical partners in protection against acute cell injury. Critical Care Medicine, 30, S43-S50. doi:10.1097/00003246-200201001-00006
[21] Garigan, D., Hsu, A.L., Fraser, A.G., Kamath, R.S., Ahringer, J. and Kenyon, C. (2002) Genetic analysis of tissue aging in Caenorhabditis elegans: A role for heatshock factor and bacterial proliferation. Genetics, 161, 1101-1112.
[22] Sorensen, J.G., Kristensen, T.N., Kristensen, K.V. and Loeschcke, V. (2007) Sex specific effects of heat induced hormesis in Hsf-deficient Drosophila melanogaster. Experimental Gerontology, 42, 1123-1129. doi:10.1016/j.exger.2007.09.001
[23] Xiao, X., Zuo, X., Davis, A.A., McMillan, D.R., Curry, B.B., Richardson, J.A. and Benjamin, I.J. (1999) HSF1 is required for extra-embryonic development, postnatal growth and protection during inflammatory responses in mice. EMBO Journal, 18, 5943-5952. doi:10.1093/emboj/18.21.5943
[24] Kourennaia, O.V. and Dehaseth, P.L. (2007) Substitution of a highly conserved histidine in the Escherichia coli heat shock transcription factor, sigma32, affects promoter utilization in vitro and leads to overexpression of the biofilm-associated flu protein in vivo. Journal of Bacteriology, 189, 8430-8436. doi:10.1128/JB.01197-07
[25] Charng, Y.Y., Liu, H.C., Liu, N.Y., Chi, W.T., Wang, C.N., Chang, S.H. and Wang, T.T. (2007) A heat-inducible transcription factor, HsfA2, is required for extension of acquired thermotolerance in Arabidopsis. Plant Physiology, 143, 251-262. doi:10.1104/pp.106.091322
[26] Nishizawa, A., Yabuta, Y., Yoshida, E., Maruta, T., Yoshimura, K. and Shigeoka, S. (2006) Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress. Plant Journal, 48, 535-547. doi:10.1111/j.1365-313X.2006.02889.x
[27] Zhu, B., Ye, C., Lu, H., Chen, X., Chai, G., Chen, J. and Wang, C. (2006) Identification and characterization of a novel heat shock transcription factor gene, GmHsfA1, in soybeans (Glycine max). Journal of Plant Research, 119, 247-256. doi:10.1007/s10265-006-0267-1
[28] Gasch, A.P., Spellman, P.T., Kao, C.M., Carmel-Harel, O., Eisen, M.B., Storz, G., Botstein, D. and Brown, P.O. (2000) Genomic expression programs in the response of yeast cells to environmental changes. Molecular Biology of the Cell, 11, 4241-4257.
[29] Tucker, C.L. and Fields, S. (2004) Quantitative genome-wide analysis of yeast deletion strain sensitivities to oxidative and chemical stress. Comparative and Functional Genomics, 5, 216-224. doi:10.1002/cfg.391
[30] Tan, S.X., Teo, M., Lam, Y.T., Dawes, I.W. and Perrone, G.G. (2009) Cu, Zn superoxide dismutase and NADP(H) homeostasis are required for tolerance of endoplasmic reticulum stress in Saccharomyces cerevisiae. Molecular Biology of the Cell, 20, 1493-1508. doi:10.1091/mbc.E08-07-0697
[31] Han, D., Antunes, F., Canali, R., Rettori, D. and Cadenas, E. (2003) Voltage-dependent anion channels control the release of the superoxide anion from mitochondria to cytosol. The Journal of Biological Chemistry, 278, 5557-5563. doi:10.1074/jbc.M210269200
[32] Hahn, J.S., Hu, Z., Thiele, D.J. and Iyer, V.R. (2004) Genome-wide analysis of the biology of stress responses through heat shock transcription factor. Molecular and Cellular Biology, 24, 5249-5256. doi:10.1128/MCB.24.12.5249-5256.2004
[33] Yamamoto, A., Mizukami, Y. and Sakurai, H. (2005) Identification of a novel class of target genes and a novel type of binding sequence of heat shock transcription factor in Saccharomyces cerevisiae. The Journal of Biological Chemistry, 280, 11911-11919. doi:10.1074/jbc.M411256200
[34] Gong, Y., Kakihara, Y., Krogan, N., Greenblatt, J., Emili, A., Zhang, Z. and Houry, W.A. (2009) An atlas of chaperone-protein interactions in Saccharomyces cerevisiae: Implications to protein folding pathways in the cell. Molecular Systems Biology, 5, 275. doi:10.1038/msb.2009.26
[35] Hartl, F.U. and Hayer-Hartl, M. (2002) Molecular chaperones in the cytosol: From nascent chain to folded protein. Science, 295, 1852-1858. doi:10.1126/science.1068408
[36] Haslbeck, M., Miess, A., Stromer, T., Walter, S. and Buchner, J. (2005) Disassembling protein aggregates in the yeast cytosol. The cooperation of Hsp26 with Ssa1 and Hsp104. The Journal of Biological Chemistry, 280, 23861-23868. doi:10.1074/jbc.M502697200
[37] Zhao, R., Davey, M., Hsu, Y.C., Kaplanek, P., Tong, A., Parsons, A.B., Krogan, N., Cagney, G., Mai, D., Greenblatt, J., Boone, C., Emili, A. and Houry W.A. (2005) Navigating the chaperone network: An integrative map of physical and genetic interactions mediated by the hsp90 chaperone. Cell, 120, 715-727. doi:10.1016/j.cell.2004.12.024
[38] Mayr, C., Richter, K., Lilie, H. and Buchner, J. (2000) Cpr6 and Cpr7, two closely related Hsp90-associated immunophilins from Saccharomyces cerevisiae, differ in their functional properties. The Journal of Biological Chemistry, 275, 34140-34146. doi:10.1074/jbc.M005251200
[39] Wegele, H., Haslbeck, M., Reinstein, J. and Buchner, J. (2003) Sti1 is a novel activator of the Ssa proteins. The Journal of Biological Chemistry, 278, 25970-25976. doi:10.1074/jbc.M301548200
[40] Chu, F., Maynard, J.C., Chiosis, G., Nicchitta, C.V. and Burlingame, A.L. (2006) Identification of novel quaternary domain interactions in the Hsp90 chaperone, GRP94. Protein Science, 15, 1260-1269. doi:10.1110/ps.052065106

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