Regulation of the gene encoding glutathione synthetase from the fission yeast.
The fission yeast cells that contained the cloned glutathione synthetase (GS) gene showed 1.4-fold higher glutathione (GSH) 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 multicopy-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 microM) than the cells that harbored the YEp357R vector. The 506 bp upstream sequence from the translational initiation point and N-terminal 8 amino acid-coding region were fused into the promoterless 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.
Kim
Su-Jung
SJ
Division of Life Sciences, Kangwon National University, Chuncheon 200-701, Korea.
Shin
Youn Hee
YH
Kim
Kyunghoon
K
Park
Eun-Hee
EH
Sa
Jae-Hoon
JH
Lim
Chang-Jin
CJ
D019328:Buthionine Sulfoximine / Q000494:pharmacology; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005981:Glutathione Synthase / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D007763:Lac Operon / Q000235:genetics; D019216:Metals, Heavy / Q000494:pharmacology; D008969:Molecular Sequence Data; D009599:Nitroprusside / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D013997:Time Factors; D024483:Vitamin K 3 / Q000494:pharmacology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000187:drug effects
D019216:Metals, Heavy; D011993:Recombinant Fusion Proteins; D009599:Nitroprusside; D019328:Buthionine Sulfoximine; D024483:Vitamin K 3; D026423:S-Nitroso-N-Acetylpenicillamine; D001616:beta-Galactosidase; D005981:Glutathione Synthase