Characterization and regulation of the γ-glutamyl transpeptidase gene from the fission yeast Schizosaccharomyces pombe

2004 ◽  
Vol 50 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Hey-Jung Park ◽  
Hye-Won Lim ◽  
Kanghwa Kim ◽  
Il-Han Kim ◽  
Eun-Hee Park ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Rattus Norvegicus ◽  
Genomic Dna ◽  
Homo Sapiens ◽  
Shuttle Vector ◽  
Mrna Level ◽  
Exponential Phase ◽  
Chromosomal Dna ◽  
Glutamyl Transpeptidase

The structural gene for the putative γ-glutamyl transpeptidase (GGT) was isolated from the chromosomal DNA of the fission yeast Schizosaccharomyces pombe. The determined sequence contained 3324 bp and encoded the predicted 630 amino acid sequence of GGT, which resembles counterparts in Homo sapiens, Rattus norvegicus, Saccharomyces cerevisiae, and Escherichia coli. The S. pombe cells harboring the cloned GGT gene showed about twofold higher GGT activity in the exponential phase than the cells harboring the vector only, indicating that the cloned GGT gene was functional. To monitor the expression of the S. pombe GGT gene, we fused the fragment 1085 bp upstream of the cloned GGT gene into the promoterless β-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pGT98. The synthesis of β-galactosidase from the fusion plasmid in S. pombe cells was enhanced by treatments with NO-generating sodium nitroprusside (SN), L-buthionine-(S,R)-sulfoximine (BSO), and glycerol. The GGT mRNA level in the S. pombe cells was increased by SN and BSO. Involvement of Pap1 in the induction of the GGT gene by SN and BSO was observed.Key words: fission yeast, genomic DNA, γ-glutamyl transpeptidase, Pap1, regulation, Schizosaccharomyces pombe.

Characterization of a second gene encoding γ-glutamyl transpeptidase from Schizosaccharomyces pombe

2005 ◽  
Vol 51 (3) ◽  
pp. 269-275 ◽  
Author(s):  
Hey-Jung Park ◽  
Jeong-Su Moon ◽  
Hong-Gyum Kim ◽  
Il-Han Kim ◽  
Kanghwa Kim ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Genomic Dna ◽  
Homo Sapiens ◽  
Shuttle Vector ◽  
Mrna Level ◽  
Gene Encoding ◽  
Oxidative Stresses ◽  
Solid Media ◽  
Glutamyl Transpeptidase

The first gene encoding γ-glutamyl transpeptidase (GGTI) of the fission yeast has previously been characterized, and its expression was found to be regulated by various oxidative stress-inducing agents. In this work, a second gene, encoding GGTII, was cloned and characterized from the fission yeast Schizosaccharomyces pombe. The structural gene encoding GGTII was amplified from the genomic DNA of the fission yeast and ligated into the shuttle vector pRS316 to generate the recombinant plasmid pPHJ02. The determined sequence contains 3040 bp and is able to encode the putative 611 amino acid sequence of GGTII, which resembles the counterparts of Saccharomyces cerevisiae, Homo sapiens, Rattus norvegicus, and Escherichia coli. The DNA sequence also contains 940-bp upstream and 289-bp downstream regions of the GGTII gene. The Schizosaccharomyces pombe cells harboring plasmid pPHJ02 showed about 4-fold higher GGT activity in the exponential phase than the cells harboring the vector only, indicating that the cloned GGTII gene is functional. The S. pombe cells containing the cloned GGTII gene were found to contain higher levels of both intracellular glutathione (GSH) content and GSH uptake. The S. pombe cells harboring plasmid pPHJ02 showed increased survival on solid media containing hydrogen peroxide, diethylmaleate, aluminum chloride, cadmium chloride, or mercuric chloride. The GGTII mRNA level was significantly elevated by treatment with GSH-depleting diethylmaleate. These results imply that the S. pombe GGTII gene produces functional GGTII protein and is involved in the response to oxidative stresses in S. pombe cells.Key words: fission yeast, genomic DNA, γ-glutamyl transpeptidase, regulation, Schizosaccharomyces pombe, stress response.

scholarly journals DNA polymerase ε encoded by cdc20 + is required for chromosomal DNA replication in the fission yeast Schizosaccharomyces pombe

1998 ◽  
Vol 3 (2) ◽  
pp. 99-110 ◽  
Author(s):  
Akio Sugino ◽  
Takeshi Ohara ◽  
Josef Sebastian ◽  
Naomi Nakashima ◽  
Hiroyuki Araki
Keyword(s):  
Dna Replication ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dna Polymerase ◽  
Chromosomal Dna

scholarly journals Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe.

Genetics ◽  
1994 ◽  
Vol 136 (3) ◽  
pp. 849-856 ◽  
Author(s):  
J B Keeney ◽  
J D Boeke
Keyword(s):  
Homologous Recombination ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Gene Conversion ◽  
Blot Analysis ◽  
Genomic Dna ◽  
Plasmid Vector ◽  
Homologous Integration ◽  
Targeted Integration ◽  
Chromosomal Loci

Abstract Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1+ and ura4+ genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1+ or ura4+. Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1+ and ura4+ plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.

Overexpression of a metacaspase gene stimulates cell growth and stress response inSchizosaccharomyces pombe

2007 ◽  
Vol 53 (8) ◽  
pp. 1016-1023 ◽  
Author(s):  
Hye-Won Lim ◽  
Su-Jung Kim ◽  
Eun-Hee Park ◽  
Chang-Jin Lim
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Stress Response ◽  
Cell Growth ◽  
Schizosaccharomyces Pombe ◽  
Sodium Nitroprusside ◽  
Fission Yeast ◽  
Recombinant Plasmid ◽  
Mrna Level ◽  
Yeast Cells ◽  
Yeast Saccharomyces Cerevisiae

A unique gene named pca1+, encoding a metacaspase, was cloned from the fission yeast Schizosaccharomyces pombe and was used to create a recombinant plasmid, pPMC. The metacaspase mRNA level was markedly elevated in the fission yeast cells harboring the plasmid pPMC. Overexpressed Pca1+appeared to stimulate the growth of the fission yeast cells instead of arresting their growth. Its expression was enhanced by stress-inducing agents such as H2O2, sodium nitroprusside, and CdCl2, and it conferred cytoprotection, especially against CdCl2. However, such protection was not reproducible in the budding yeast Saccharomyces cerevisiae harboring pPMC. Taken together, these results propose that Pca1+may be involved in the growth and stress response of the fission yeast.

Stress-dependent regulation of Pbh1, a BIR domain-containing protein, in the fission yeast

2006 ◽  
Vol 52 (12) ◽  
pp. 1261-1265 ◽  
Author(s):  
Nam-Chul Cho ◽  
Hyun-Jung Kang ◽  
Hye-Won Lim ◽  
Byung-Chul Kim ◽  
Eun-Hee Park ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nitrosative Stress ◽  
Nitrogen Starvation ◽  
Fusion Gene ◽  
Shuttle Vector ◽  
Upstream Region ◽  
Basal Expression ◽  
Stress Dependent ◽  
Bir Domain

To elicit the physiological roles of Pbh1, a baculoviral IAP repeat (BIR) domain-containing protein, in Schizosaccharomyces pombe, we investigated if Pbh1 expression is regulated by stress. The upstream region (1221 bp) of the pbh1 gene was fused into the promoterless β-galactosidase gene of the shuttle vector YEp367R, and the resultant fusion plasmid was named pPbh04. The synthesis of β-galactosidase from the pbh1-lacZ fusion gene was markedly enhanced by sodium nitroprusside (SNP) generating nitric oxide. The basal expression of the pbh1 gene required the presence of Pap1. Pap1 also mediated the induction of the pbh1 gene by SNP and nitrogen starvation. Pap1-dependent induction of the pbh1 gene by SNP was confirmed by the enhanced level of the pbh1 mRNA in Pap1-positive cells but not in Pap1-negative cells. Taken together, it was demonstrated that the pbh1 genes are positively regulated by nitrosative and nitrogen starvation stresses in Pap1-dependent manner.Key words: fission yeast, nitrosative stress, nutritional stress, nitrogen starvation, Pap1, Pbh1, regulation, Schizosaccharomyces pombe.

scholarly journals Upstream – News in Genomics

2002 ◽  
Vol 3 (3) ◽  
pp. 221-225
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ovarian Cancer ◽  
Oryza Sativa ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Proteomic Analysis ◽  
Large Scale ◽  
Protein Complexes ◽  
The Other ◽  
Blood Samples

In recent months a bumper crop of genomes has been completed, including the fission yeast (Schizosaccharomyces pombe) and rice (Oryza sativa). Two large-scale studies ofSaccharomyces cerevisiaeprotein complexes provided a picture of the eukaryotic proteome as a network of complexes. Amongst the other stories of interest was a demonstration that proteomic analysis of blood samples can be used to detect ovarian cancer, perhaps even as early as stage I.

scholarly journals Analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe.

1986 ◽  
Vol 83 (21) ◽  
pp. 8253-8257 ◽  
Author(s):  
L. Clarke ◽  
H. Amstutz ◽  
B. Fishel ◽  
J. Carbon
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Centromeric Dna
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