A Protein Differential Analysis of Cell Wall in Saccharomyces cerevisiae Under Different Temperatures

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.029

Abstract

Abstract: Living organisms undergo a series of stresses from abiotic environment, and these stresses will affect the changes of genetic transcription and the expression of protein for quick adaptation to the changing environment. Two-dimensional electrophoresis and mass spectrometry were used to investigate the effects of high-temperature stresses on the proteome of cell wall in Saccharomyces cerevisiae. The results showed that, in S. cerevisiae FFC2146 under high temperature, proteins of cell walls had new Ssa2 and small molecules guanosine triphosphatase, inorganic pyrophosphatase was up-regulated, pyruvate kinase was deficient or even disappeared, and the expression of glucose-6-phosphate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase was down-regulated. These results indicated that the heat shock protein Ssa2 protected cell walls to be intact at a high temperature, therefore the cells grew and reproduced continuously；EMP glycolytic pathway of S. cerevisiae under heat stress was blocked, glycolytic pathway turned HMP pathway by transketolase, enough energy was obtained to maintain normal metabolism of cell.

Key words: Saccharomyces cerevisiae, two-dimensional electrophoresis, heat shock protein

Gu Yue, Piao Yongzhe, Du Wei, Wang Xiaoyu, Wang Chunyan. A Protein Differential Analysis of Cell Wall in Saccharomyces cerevisiae Under Different Temperatures[J]. Biotechnology Bulletin, 2015, 31(12): 200-206.

References

[1] Stanley D, Bandara A, Fraser S, et al. The ethanol stress response and ethanol tolerance of Saccharomyces cerevisiae[J]. Appl Microbiol, 2010, 109(1):13-24.
[2]Suutari M, Liukkonen K, Laakso S. Temperature adaptation in yeast:the role of fatty acids[J]. Gen Appl Microbiol, 1990, 136(1):1469-1474.
[3]Benschoter AS, Ingram O. Thermal tolerance of Zymomonas mobilis:Temperature-induced changes in membrane composition[J]. Appl Environ Microb, 1986, 6(4):1278-1284.
[4]Lindquist S, Kim G. Heat shock protein 104 expression is sufficient for thermotolerance in yeast[J]. Proc Natl Acad Sci USA, 1996, 93(3):5301-5306.
[5]Hottiger T, Virgilio C, Hall M N, et al. The role of trehalose synthesis for the acquisition of thermotolerance in yeast[J]. Eur J Biochem, 2005, 219(10):187-193.
[6]Belloch C, Orlic S, Barrio E, et al. Fermentative stress adaptation of hybrids within the Saccharomyces sensu stricto complex[J]. Int J Food Microbiol, 2008, 122(3):188-195.
[7] Griffin TJ, Aebersold R, Chem JB. Advances in Proteome Analysis by Mass Spectrometry[J]. Molecular and Cellular Biology, 2001, 276(2):45497-45500.
[8] 姚继兵, 祖国仁, 朴永哲, 等. 不同传代次数的酿酒酵母细胞壁蛋白组学分析[J]. 微生物学通报, 2013, 40(11):1962-1969.
[9]季杨杨, 朴永哲, 袁方, 等. 不同传代次数酿酒酵母细胞内蛋白质组学分析[J]. 食品科技, 2014, 9(7):26-30.
[10]程君升, 毛开荣, 丁家波, 等. 微量Bradford法测定提纯禽结核菌素蛋白含量[J]. 中国兽药杂志, 2007, 41(6):9-11.
[11]Bienz M, Pelham HR. Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter[J]. Cell, 1986, 45(3), 753-760.
[12]Hartl FU. Molecular chaperones in cellular protein folding[J]. Nature, 1996, 3(8):571-580.
[13]Mayer M, Bukau B. Hsp70 chaperones:cellular functions and molecular mechanism[J]. Cell Mol Life Sci, 2005, 62(6):670-684.
[14] Lopez JL, Chaffin WL. Member of the Hsp70 family of proteins in the cell wall of Saccharomyces cerevisiae[J]. Journal of Bacteriology, 1996, 180(2):4724-4726.
[15]Sharma D, Masion CN. Functionally redundant isoforms of a yeast Hsp70 chaperone subfamily have different antiprion effects[J]. Genetics, 2008, 179(3):1301-1311.
[16]Harold FM, Bacteriol H. Inorganic polyphosphates in biology:structure, metabolism, and function[J]. Bacteriol Rev, 1966, 30(4):772-778.
[17] Yoon HS, Kim SY, Kim S. Stess response of plant H⁺-PPase-expressing transgenic Escherichia coli and Saccharomyces cerevisiae:a potentially useful mechanism for the development of stress-tolerant organisms[J]. J App l Genetics, 2013, 3(54):129-133.
[18] Terkeltaub RA. Inorganic pyrophosphate generation and disposition in pathophysiology[J]. Physiol Cell Physiol, 2001, 281(1):C1-C11.
[19]Larsson C, Pahlman I, Gustafsson L. The importance of ATP as a regulator of glycolytic flux in Saccharomyces cerevisiae[J]. Yeast, 2000, 16(3):797-809.
[20] Giardina BJ, Chiang HL. Fructose-1, 6-bisphosphatase, malate de-hydrogenase, isocitrate lyase, phosphoenolpyruvate carboxykinase, glyceraldehyde-3-phosphate dehydrogenase, and cyclophilin a are secreted in Saccharomyces cerevisiae grown in low glucose[J]. Communicative&Integrative Biology, 2013, 6(6):234-238.
[21] Pardo M, Monteoliva L, Pla J, et al. Two-dimensional analysis of pr-oteins secreted by Saccharomyces cerevisiae regenerating protoplasts a novel approach to study the cell wall[J]. Yeast, 1999, 5(43):459-472.
[22] Endo T, Mitsui S, Nakai M, et al. Binding of tochondrial presequen-ces to yeast cytosolic heat shock protein70 depends on the a mphip-hilicity of the presequence[J]. Journal of Biological Chemistry, 1996, 271(8):4161-4167.
[23]Baudin A. simple and efficient method of direct gene deletion in Saccharomyces cerevisiae[J]. Nucleic Acids Res, 1993, 21(2):3329-3330.