Abstract
Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi-functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of beta-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of beta-galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of beta-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Basic-Leucine Zipper Transcription Factors / metabolism
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Basic-Leucine Zipper Transcription Factors / pharmacology
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Dose-Response Relationship, Drug
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Gene Expression Regulation*
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Glutaredoxins
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Molecular Sequence Data
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Nitroprusside / metabolism*
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Nitroprusside / pharmacology
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Osmotic Pressure*
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Oxidative Stress / genetics
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Oxidative Stress / physiology
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Oxidoreductases / genetics*
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Oxidoreductases / metabolism
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Oxidoreductases / physiology
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Pancreatitis-Associated Proteins
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Reactive Oxygen Species / metabolism
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Reactive Oxygen Species / pharmacology
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Schizosaccharomyces / genetics*
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Schizosaccharomyces / metabolism
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Schizosaccharomyces / physiology
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Schizosaccharomyces pombe Proteins / metabolism
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Schizosaccharomyces pombe Proteins / pharmacology
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Sequence Alignment
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Time Factors
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beta-Galactosidase / genetics
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beta-Galactosidase / metabolism
Substances
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Basic-Leucine Zipper Transcription Factors
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Glutaredoxins
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Pancreatitis-Associated Proteins
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Pap1 protein, S pombe
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REG3A protein, human
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RNA, Messenger
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Reactive Oxygen Species
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Schizosaccharomyces pombe Proteins
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Nitroprusside
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Oxidoreductases
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beta-Galactosidase