The gene encoding gamma-glutamyl transpeptidase II in the fission yeast is regulated by oxidative and metabolic stress.
gamma-Glutamyl transpeptidase (GGT, EC 2.3.2.2.) catalyzes the transfer of the gamma-glutamyl moiety from gamma-glutamylcontaining compounds, notably glutathione (GSH), to acceptor amino acids and peptides. A second gene (GGTII) encoding GGT was previously isolated and characterized from the fission yeast Schizosaccharomyces pombe. In the present work, the GGTII-lacZ fusion gene was constructed and used to study the transcriptional regulation of the S. pombe GGTII gene. The synthesis of beta-galactosidase from the GGTII-lacZ fusion gene was significantly enhanced by NO-generating SNP and hydrogen peroxide in the wildtype yeast cells. The GGTII mRNA level was increased in the wild-type S. pombe cells treated with SNP. However, the induction by SNP was abolished in the Pap1-negative S. pombe cells, implying that the induction by SNP of GGTII is mediated by Pap1. Fermentable carbon sources, such as glucose (at low concentrations), lactose and sucrose, as a sole carbon source, enhanced the synthesis of beta-galactosidase from the GGTII-lacZ fusion gene in wildtype KP1 cells but not in Pap1-negative cells. Glycerol, a non-fermentable carbon source, was also able to induce the synthesis of beta-galactosidase from the fusion gene, but other non-fermentable carbon sources such as acetate and ethanol were not. Transcriptional induction of the GGTII gene by fermentable carbon sources was also confirmed by increased GGTII mRNA levels in the yeast cells grown with them. Nitrogen starvation was also able to induce the synthesis of beta-galactosidase from the GGTII-lacZ fusiongene in a Pap1-dependent manner. On the basis of the results, it is concluded that the S. pombe GGTII gene is regulated by oxidative and metabolic stress.
Kang
Hyun-Jung
HJ
Division of Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Korea.
Kim
Byung-Chul
BC
Park
Eun-Hee
EH
Ahn
Kisup
K
Lim
Chang-Jin
CJ
D000818:Animals; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D020030:Nitric Oxide Donors / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism; D009599:Nitroprusside / Q000378:metabolism; D016877:Oxidants / Q000378:metabolism; D018384:Oxidative Stress; D000073718:Pancreatitis-Associated Proteins; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D005723:gamma-Glutamyltransferase / Q000235:genetics / Q000378:metabolism
D050976:Basic-Leucine Zipper Transcription Factors; D005656:Fungal Proteins; D020030:Nitric Oxide Donors; D016877:Oxidants; D000073718:Pancreatitis-Associated Proteins; C105096:Pap1 protein, S pombe; C000612141:REG3A protein, human; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D009599:Nitroprusside; D002244:Carbon; D006861:Hydrogen Peroxide; D005723:gamma-Glutamyltransferase; D005947:Glucose; D009584:Nitrogen