Distinct Redox Regulation in Sub-Cellular Compartments in Response to Various Stress Conditions in Saccharomyces cerevisiae

被引:43
作者
Ayer, Anita [1 ]
Sanwald, Julia [1 ]
Pillay, Bethany A. [1 ]
Meyer, Andreas J. [2 ]
Perrone, Gabriel G. [3 ]
Dawes, Ian W. [1 ]
机构
[1] Univ New S Wales, Sydney, NSW, Australia
[2] Univ Bonn, INRES Chem Signalling, Bonn, Germany
[3] Univ Western Sydney, Penrith, NSW 1797, Australia
来源
PLOS ONE | 2013年 / 8卷 / 06期
基金
澳大利亚研究理事会;
关键词
MITOCHONDRIAL INTERMEMBRANE SPACE; INDUCED CELL-DEATH; OXIDATIVE-STRESS; TRANSCRIPTIONAL RESPONSE; HYDROGEN-PEROXIDE; YEAST-CELLS; GLUTATHIONE; PROTEIN; SHOCK; PH;
D O I
10.1371/journal.pone.0065240
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Responses to many growth and stress conditions are assumed to act via changes to the cellular redox status. However, direct measurement of pH-adjusted redox state during growth and stress has never been carried out. Organellar redox state (E-GSH) was measured using the fluorescent probes roGFP2 and pHluorin in Saccharomyces cerevisiae. In particular, we investigated changes in organellar redox state in response to various growth and stress conditions to better understand the relationship between redox-, oxidative-and environmental stress response systems. E-GSH values of the cytosol, mitochondrial matrix and peroxisome were determined in exponential and stationary phase in various media. These values (-340 to -350 mV) were more reducing than previously reported. Interestingly, sub-cellular redox state remained unchanged when cells were challenged with stresses previously reported to affect redox homeostasis. Only hydrogen peroxide and heat stress significantly altered organellar redox state. Hydrogen peroxide stress altered the redox state of the glutathione disulfide/glutathione couple (GSSG, 2H(+)/2GSH) and pH. Recovery from moderate hydrogen peroxide stress was most rapid in the cytosol, followed by the mitochondrial matrix, with the peroxisome the least able to recover. Conversely, the bulk of the redox shift observed during heat stress resulted from alterations in pH and not the GSSG, 2H(+)/2GSH couple. This study presents the first direct measurement of pH-adjusted redox state in sub-cellular compartments during growth and stress conditions. Redox state is distinctly regulated in organelles and data presented challenge the notion that perturbation of redox state is central in the response to many stress conditions.
引用
收藏
页数:13
相关论文
共 55 条
[1]   A suite of Gateway® cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae [J].
Alberti, Simon ;
Gitler, Aaron D. ;
Lindquist, Susan .
YEAST, 2007, 24 (10) :913-919
[2]   Inhibition of sodium/proton exchange by a Rab-GTPase-activating protein regulates endosomal traffic in yeast [J].
Ali, R ;
Brett, CL ;
Mukherjee, S ;
Rao, R .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (06) :4498-4506
[3]   A Genome-Wide Screen in Yeast Identifies Specific Oxidative Stress Genes Required for the Maintenance of Sub-Cellular Redox Homeostasis [J].
Ayer, Anita ;
Fellermeier, Sina ;
Fife, Christopher ;
Li, Simone S. ;
Smits, Gertien ;
Meyer, Andreas J. ;
Dawes, Ian W. ;
Perrone, Gabriel G. .
PLOS ONE, 2012, 7 (09)
[4]   The critical role of glutathione in maintenance of the mitochondrial genome [J].
Ayer, Anita ;
Tan, Shi-Xiong ;
Grant, Chris M. ;
Meyer, Andreas J. ;
Dawes, Ian W. ;
Perrone, Gabriel G. .
FREE RADICAL BIOLOGY AND MEDICINE, 2010, 49 (12) :1956-1968
[5]   Measuring intracellular redox conditions using GFP-based sensors [J].
Bjornberg, Olof ;
Ostergaard, Henrik ;
Winther, Jakob R. .
ANTIOXIDANTS & REDOX SIGNALING, 2006, 8 (3-4) :354-361
[6]   The yeast endosomal Na+(K+)/H+ exchanger Nhx1 regulates cellular pH to control vesicle trafficking [J].
Brett, CL ;
Tukaye, DN ;
Mukherjee, S ;
Rao, RJ .
MOLECULAR BIOLOGY OF THE CELL, 2005, 16 (03) :1396-1405
[7]   The cell cycle is a redox cycle: Linking phase-specific targets to cell fate [J].
Burhans, William C. ;
Heintz, Nicholas H. .
FREE RADICAL BIOLOGY AND MEDICINE, 2009, 47 (09) :1282-1293
[8]   Remodeling of yeast genome expression in response to environmental changes [J].
Causton, HC ;
Ren, B ;
Koh, SS ;
Harbison, CT ;
Kanin, E ;
Jennings, EG ;
Lee, TI ;
True, HL ;
Lander, ES ;
Young, RA .
MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (02) :323-337
[9]   ISOLATION OF THE CATALASE-A GENE OF SACCHAROMYCES-CEREVISIAE BY COMPLEMENTATION OF THE CTA1 MUTATION [J].
COHEN, G ;
FESSL, F ;
TRACZYK, A ;
RYTKA, J ;
RUIS, H .
MOLECULAR & GENERAL GENETICS, 1985, 200 (01) :74-79
[10]   INDUCIBILITY OF THE RESPONSE OF YEAST-CELLS TO PEROXIDE STRESS [J].
COLLINSON, LP ;
DAWES, IW .
JOURNAL OF GENERAL MICROBIOLOGY, 1992, 138 :329-335