A genetic investigation of the essential role of glutathione - Mutations in the proline biosynthesis pathway are the only suppressors of glutathione auxotrophy in yeast

被引:70
作者
Spector, D
Labarre, J
Toledano, MB
机构
[1] CEA, Serv Biochim & Genet Mol, F-91191 Gif Sur Yvette, France
[2] Rutgers State Univ, Coll Pharm, Dept Pharmacol & Toxicol, Piscataway, NJ 08855 USA
[3] Univ Med & Dent New Jersey, Grad Sch Biomed Sci, Grad Program Mol Genet & Microbiol, Piscataway, NJ 08854 USA
关键词
D O I
10.1074/jbc.M009814200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In an attempt to elucidate the essential function of glutathione in Saccharomyces cerevisiae, we searched for suppressors of the GSH auxotrophy of Delta gsh1, a strain lacking the rate-limiting enzyme of glutathione biosynthesis. We found that specific mutations of PRO2, the second enzyme in proline biosynthesis, permitted the growth of Delta gsh1 in the absence of exogenous GSH, The suppression mechanism by alleles of PRO2 involved the biosynthesis of a trace amount of glutathione, Deletion of PRO1, the first enzyme of the proline biosynthesis pathway, or PRO2 eliminated the suppression, suggesting that gamma -glutamyl phosphate, the product of Pro1 and the physiological substrate of Pro2, is required as an obligate substrate of suppressor alleles of PRO2 for glutathione synthesis, A mutagenesis of a Delta gsh1 strain also lacking the proline pathway failed to generate any suppressor mutants under either aerobic or anaerobic conditions, confirming that glutathione is essential in yeast, This essential function is not related to DNA synthesis based on the terminal phenotype of glutathione-depleted cells or to toxic accumulation of non-native protein disulfides, Analysis of the suppressor strain demonstrates that normal glutathione levels are required for the tolerance to oxidants under acute, but not chronic stress conditions.
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收藏
页码:7011 / 7016
页数:6
相关论文
共 39 条
[1]   ISOLATION AND INITIAL CHARACTERIZATION OF GLUTATHIONE-DEFICIENT MUTANTS OF ESCHERICHIA-COLI K-12 [J].
APONTOWEIL, P ;
BERENDS, W .
BIOCHIMICA ET BIOPHYSICA ACTA, 1975, 399 (01) :10-22
[2]   Roles of the glutathione- and thioredoxin-dependent reduction systems in the Escherichia coli and Saccharomyces cerevisiae responses to oxidative stress [J].
Carmel-Harel, O ;
Storz, G .
ANNUAL REVIEW OF MICROBIOLOGY, 2000, 54 :439-461
[3]  
CHAE HZ, 1993, J BIOL CHEM, V268, P16815
[4]  
COBLENZ A, 1994, FEMS MICROBIOL REV, V14, P303, DOI 10.1111/j.1574-6976.1994.tb00103.x
[5]  
DUNCAN WSS, 1996, FEMS MICROBIOL LETT, V141, P207
[6]   Glutathione synthetase is dispensable for growth under both normal and oxidative stress conditions in the yeast Saccharomyces cerevisiae due to an accumulation of the dipeptide gamma-glutamylcysteine [J].
Grant, CM ;
MacIver, FH ;
Dawes, IW .
MOLECULAR BIOLOGY OF THE CELL, 1997, 8 (09) :1699-1707
[7]  
Grant CM, 1999, MOL CELL BIOL, V19, P2650
[8]   Synthesis and role of glutathione in protection against oxidative stress in yeast [J].
Grant, CM ;
Dawes, IW .
REDOX REPORT, 1996, 2 (04) :223-229
[9]   Glutathione is an essential metabolite required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae [J].
Grant, CM ;
MacIver, FH ;
Dawes, IW .
CURRENT GENETICS, 1996, 29 (06) :511-515
[10]   Mitochondrial function is required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae [J].
Grant, CM ;
MacIver, FH ;
Dawes, IW .
FEBS LETTERS, 1997, 410 (2-3) :219-222