The Thioredoxin-Thioredoxin Reductase System Can Function in Vivo as an Alternative System to Reduce Oxidized Glutathione in Saccharomyces cerevisiae

Shi Xiong Tan, Darren Greetham, Sebastian Raeth, Chris M. Grant, Ian W. Dawes, Gabriel G. Perrone

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Cellular mechanisms that maintain redox homeostasis are crucial, providing buffering against oxidative stress. Glutathione, the most abundant low molecular weight thiol, is considered the major cellular redox buffer in most cells. To better understand how cells maintain glutathione redox homeostasis, cells of Saccharomyces cerevisiae were treated with extracellular oxidized glutathione (GSSG), and the effect on intracellular reduced glutathione (GSH) and GSSG were monitored over time. Intriguingly cells lacking GLR1 encoding the GSSG reductase in S. cerevisiae accumulated increased levels of GSH via a mechanism independent of the GSH biosynthetic pathway. Furthermore, residual NADPH-dependent GSSG reductase activity was found in lysate derived from glr1 cell. The cytosolic thioredoxin-thioredoxin reductase system and not the glutaredoxins (Grx1p, Grx2p, Grx6p, and Grx7p) contributes to the reduction of GSSG. Overexpression of the thioredoxins TRX1 or TRX2 in glr1 cells reduced GSSG accumulation, increased GSH levels, and reduced cellular glutathione E h′. Conversely, deletion of TRX1 or TRX2 in the glr1 strain led to increased accumulation of GSSG, reduced GSH levels, and increased cellular Eh′. Furthermore, it was found that purified thioredoxins can reduce GSSG to GSH in the presence of thioredoxin reductase and NADPH in a reconstituted in vitro system. Collectively, these data indicate that the thioredoxin-thioredoxin reductase system can function as an alternative system to reduce GSSG in S. cerevisiae in vivo.

Original languageEnglish
Pages (from-to)6118-6126
Number of pages9
JournalJournal of Biological Chemistry
Volume285
Issue number9
Early online date1 Dec 2009
DOIs
Publication statusPublished - 26 Feb 2010
Externally publishedYes

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Thioredoxin-Disulfide Reductase
Thioredoxins
Glutathione Disulfide
Yeast
Saccharomyces cerevisiae
Glutathione
Oxidation-Reduction
Oxidoreductases
Homeostasis
Glutaredoxins
Oxidative stress
Biosynthetic Pathways
NADP
Sulfhydryl Compounds
Buffers
Oxidative Stress
Molecular Weight
Molecular weight

Cite this

Tan, Shi Xiong ; Greetham, Darren ; Raeth, Sebastian ; Grant, Chris M. ; Dawes, Ian W. ; Perrone, Gabriel G. / The Thioredoxin-Thioredoxin Reductase System Can Function in Vivo as an Alternative System to Reduce Oxidized Glutathione in Saccharomyces cerevisiae. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 9. pp. 6118-6126.
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abstract = "Cellular mechanisms that maintain redox homeostasis are crucial, providing buffering against oxidative stress. Glutathione, the most abundant low molecular weight thiol, is considered the major cellular redox buffer in most cells. To better understand how cells maintain glutathione redox homeostasis, cells of Saccharomyces cerevisiae were treated with extracellular oxidized glutathione (GSSG), and the effect on intracellular reduced glutathione (GSH) and GSSG were monitored over time. Intriguingly cells lacking GLR1 encoding the GSSG reductase in S. cerevisiae accumulated increased levels of GSH via a mechanism independent of the GSH biosynthetic pathway. Furthermore, residual NADPH-dependent GSSG reductase activity was found in lysate derived from glr1 cell. The cytosolic thioredoxin-thioredoxin reductase system and not the glutaredoxins (Grx1p, Grx2p, Grx6p, and Grx7p) contributes to the reduction of GSSG. Overexpression of the thioredoxins TRX1 or TRX2 in glr1 cells reduced GSSG accumulation, increased GSH levels, and reduced cellular glutathione E h′. Conversely, deletion of TRX1 or TRX2 in the glr1 strain led to increased accumulation of GSSG, reduced GSH levels, and increased cellular Eh′. Furthermore, it was found that purified thioredoxins can reduce GSSG to GSH in the presence of thioredoxin reductase and NADPH in a reconstituted in vitro system. Collectively, these data indicate that the thioredoxin-thioredoxin reductase system can function as an alternative system to reduce GSSG in S. cerevisiae in vivo.",
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The Thioredoxin-Thioredoxin Reductase System Can Function in Vivo as an Alternative System to Reduce Oxidized Glutathione in Saccharomyces cerevisiae. / Tan, Shi Xiong; Greetham, Darren; Raeth, Sebastian; Grant, Chris M.; Dawes, Ian W.; Perrone, Gabriel G.

In: Journal of Biological Chemistry, Vol. 285, No. 9, 26.02.2010, p. 6118-6126.

Research output: Contribution to journalArticle

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