Stress-dependent regulation of a monothiol glutaredoxin gene from Schizosaccharomyces pombe.
Glutaredoxin (Grx) is a small, heat-stable protein acting as a multi-functional glutathione-dependent disulfide oxidoreductase. In this work, a gene encoding the monothiol glutaredoxin Grx4 was cloned from the genomic DNA of the fission yeast Schizosaccharomyces pombe. The determined DNA sequence carries 1706 bp, which is able to encode the putative 244 amino acid sequence of Grx with 27 099 Da. It does not contain an intron, and the sequence CGFS is found in the active site. Grx activity was increased 1.46-fold in S. pombe cells harboring the cloned Grx4 gene, indicating that the Grx4 gene is in vivo functioning. Although aluminum, cadmium, and hydrogen peroxide marginally enhanced the synthesis of beta-galactosidase from the Grx4-lacZ fusion gene, NO-generating sodium nitroprusside (0.5 mmol/L and 1.0 mmol/L) and potassium chloride (0.2 mol/L and 0.5 mol/L) significantly enhanced it. The Grx4 mRNA level was also enhanced after the treatment with sodium nitroprusside and potassium chloride. The synthesis of beta-galactosidase from the Grx4-lacZ gene was increased by fermentable carbon sources, such as glucose (lower than 2%) and sucrose, but not by nonfermentable carbon sources such as acetate and ethanol. The basal expression of the S. pombe Grx4 gene did not depend on the presence of Pap1. These results imply that the S. pombe monothiol Grx4 gene is genuinely functional and regulated by a variety of stresses.
Kim
Hong-Gyum
HG
Division of Life Sciences, Kangwon National University, Colege of Natural Sciences, Korea.
Kim
Byung-Chul
BC
Park
Eun-Hee
EH
Lim
Chang-Jin
CJ
D000595:Amino Acid Sequence; D050976:Basic-Leucine Zipper Transcription Factors; D015966:Gene Expression Regulation, Fungal; D054477:Glutaredoxins; D018869:Heat-Shock Response; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009599:Nitroprusside / Q000494:pharmacology; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000073718:Pancreatitis-Associated Proteins; D011189:Potassium Chloride / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism
D050976:Basic-Leucine Zipper Transcription Factors; D054477:Glutaredoxins; D000073718:Pancreatitis-Associated Proteins; C105096:Pap1 protein, S pombe; C000612141:REG3A protein, human; D029702:Schizosaccharomyces pombe Proteins; D013438:Sulfhydryl Compounds; D009599:Nitroprusside; D011189:Potassium Chloride; D010088:Oxidoreductases