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KMID : 0578320070240030316
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Molecules and Cells
2007 Volume.24 No. 3 p.316 ~ p.322
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Expression, Characterization and Regulation of a Saccharomyces cerevisiae Monothiol Glutaredoxin (Grx6) Gene in Schizosaccharomyces pombe
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Lee Jae-Hoon
Kim Kyung-Hoon
Park Eun-Hee
Ahn Ki-Sup
Lim Chang-Jin
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Abstract
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Glutaredoxins (Grxs), also known as thioltransferases (TTases), are thiol oxidoreductases that regulate cellular redox state in a variety of organisms. In the budding yeast Saccharomyces cerevisiae, Grx1 and 2 are cytosolic dithiol Grxs, while Grx3, 4 and 5 are monothiol Grxs. A gene encoding a new monothiol Grx, Grx6, was cloned from the genomic DNA of S. cerevisiae by PCR. Its DNA sequence contains 1,080 bp, and encodes a putative protein of 203 amino acid residues containing Cys-Phe-Tyr-Ser at the active site. Grx6 is similar to other monothiol Grxs in the same organism and to Grx3 in the fission yeast Schizosaccharomyces pombe. and its predicted three-dimensional structure resembles that of S. pombe Grx3. S. pombe cells harboring plasmid pFGRX6 containing the Grx6 gene had about 1.3-fold elevated Grx activity in the exponential phase, and grew better than the control cells under some stressful conditions. Synthesis of β -galactosidase from a Grx6-lacZ fusion gene in S. pombe was enhanced by potassium chloride, aluminum chloride and heat (37℃) treatment. S. pombe cells harboring plasmid pFGRX6 had elevated ROS levels whereas S. pombe cells harboring extra copies of Grx3 had reduced ROS levels.
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KEYWORD
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Budding Yeast, Fission Yeast, Grx6, Induction, LacZ Fusion, Monothiol Glutaredoxin, S. cerevisiae, S. pombe, Stress Response
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