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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Regulation of the Gene Encoding Glutathione Synthetase from the Fission Yeast

  • Kim, Su-Jung (Division of Life Sciences, Kangwon National University) ;
  • Shin, Youn-Hee (Division of Life Sciences, Kangwon National University) ;
  • Kim, Kyung-Hoon (Division of Life Sciences, Kangwon National University) ;
  • Park, Eun-Hee (College of Pharmacy, Sookmyung Womens University) ;
  • Sa, Jae-Hoon (Department of Food and Drug Analysis, Kangwon Research Institute of Health and Environment) ;
  • Lim, Chang-Jin (Division of Life Sciences, Kangwon National University)
  • Received : 2002.11.09
  • Accepted : 2002.12.18
  • Published : 2003.05.31
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Abstract

The fission yeast cells that contained the cloned glutathione synthetase (GS) gene showed 1.4-fold higher glutathione (GSB) content and 1.9-fold higher GS activity than the cells without the cloned GS gene. Interestingly, $\gamma$-glutamylcysteine synthetase activity increased 2.1-fold in the S. pombe cells that contained the cloned GS gene. The S. pombe cells that harbored the multi copy-number plasmid pRGS49 (containing the cloned GS gene) showed a higher level of survival on solid media with cadmium chloride (1 mM) or mercuric chloride ($10\;{\mu}M$) than the cells that harbored the YEp357R vector. The 506 bp upstream sequence from the translational initiation point and N-terminal8 amino acid-coding region were fused into the promoteriess $\beta$-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pUGS39. Synthesis of $\beta$-galactosidase from the fusion plasmid pUGS39 was significantly enhanced by cadmium chloride and NO-generating S-nitroso-N-acetylpenicillamine (SNAP) and sodium nitroprusside (SN). It was also induced by L-buthionine-(S,R)-sulfoximine, a specific inhibitor of $\gamma$-glutamylcysteine synthetase (GCS). We also found that the expression of the S. pombe GS gene is regulated by the Atf1-Spc1-Wis1 signal pathway.

Keywords

Acknowledgement

Supported by : Korea Research Foundation

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