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.

Stress-dependent regulation of a monothiol glutaredoxin gene from Schizosaccharomyces pombe

2005 ◽  
Vol 51 (7) ◽  
pp. 613-620 ◽  
Author(s):  
Hong-Gyum Kim ◽  
Byung-Chul Kim ◽  
Eun-Hee Park ◽  
Chang-Jin Lim
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Potassium Chloride ◽  
Sodium Nitroprusside ◽  
Fusion Gene ◽  
Mrna Level ◽  
Carbon Sources ◽  
Lacz Gene ◽  
Gene Encoding ◽  
Stress Dependent

Glutaredoxin (Grx) is a small, heat-stable protein acting as a multi-functional glutathione-dependent disulfide oxidoreductase. In this work, a gene encoding the monothiol glutaredoxin Grx4 was cloned from the genomic DNA of the fission yeast Schizosaccharomyces pombe. The determined DNA sequence carries 1706 bp, which is able to encode the putative 244 amino acid sequence of Grx with 27 099 Da. It does not contain an intron, and the sequence CGFS is found in the active site. Grx activity was increased 1.46-fold in S. pombe cells harboring the cloned Grx4 gene, indicating that the Grx4 gene is in vivo functioning. Although aluminum, cadmium, and hydrogen peroxide marginally enhanced the synthesis of β-galactosidase from the Grx4-lacZ fusion gene, NO-generating sodium nitroprusside (0.5 mmol/L and 1.0 mmol/L) and potassium chloride (0.2 mol/L and 0.5 mol/L) significantly enhanced it. The Grx4 mRNA level was also enhanced after the treatment with sodium nitroprusside and potassium chloride. The synthesis of β-galactosidase from the Grx4-lacZ gene was increased by fermentable carbon sources, such as glucose (lower than 2%) and sucrose, but not by nonfermentable carbon sources such as acetate and ethanol. The basal expression of the S. pombe Grx4 gene did not depend on the presence of Pap1. These results imply that the S. pombe monothiol Grx4 gene is genuinely functional and regulated by a variety of stresses.Key words: monothiol glutaredoxin, Pap1, regulation, Schizosaccharomyces pombe, stress response.

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.

Expression of the atf1+ gene is upregulated in fission yeast under nitrosative and nutritional stresses

2009 ◽  
Vol 55 (11) ◽  
pp. 1323-1327 ◽  
Author(s):  
S.-H. Song ◽  
B.-M. Kim ◽  
C.-J. Lim ◽  
Y.S. Song ◽  
E.-H. Park
Keyword(s):  
Nitric Oxide ◽  
Schizosaccharomyces Pombe ◽  
Sodium Nitroprusside ◽  
Fission Yeast ◽  
Fusion Gene ◽  
Lacz Fusion ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
Nitrogen Depletion ◽  
Rt Pcr

This work was designed to assess regulation of the atf1+ gene in the fission yeast Schizosaccharomyces pombe under nitrosative and nutritional stresses, using the atf1+–lacZ fusion gene and RT–PCR. Nitric oxide (NO)-generating sodium nitroprusside (SNP; 10 µmol/L) and nitrogen depletion significantly enhanced synthesis of β-galactosidase from the atf1+–lacZ fusion gene in S. pombe Pap1-positive KP1 cells, but not in S. pombe Pap1-negative TP108-3C cells. SNP (10 µmol/L) and nitrogen depletion also caused a significant increase in atf1+ mRNA levels in Pap1-positive cells, but not in Pap1-negative cells. Depletion of glucose marginally increased synthesis of β-galactosidase from the fusion gene in S. pombe Pap1-positive cells. Taken together, the S. pombe atf1+ gene is upregulated by nitrosative and nutritional stresses on a transcriptional level, possibly via the mediation of Pap1.

scholarly journals The S. pombe CDK5 ortholog Pef1 regulates sexual differentiation through control of the TORC1 pathway and autophagy

2020 ◽  
Vol 133 (17) ◽  
pp. jcs247817
Author(s):  
Shinya Matsuda ◽  
Ushio Kikkawa ◽  
Haruka Uda ◽  
Akio Nakashima
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Sexual Differentiation ◽  
Environmental Changes ◽  
Nitrogen Starvation ◽  
General Strategy ◽  
Positive Regulation ◽  
Starvation Response ◽  
Double Deletion ◽  
Early Phases

ABSTRACTIn Schizosaccharomyces pombe, a general strategy for survival in response to environmental changes is sexual differentiation, which is triggered by TORC1 inactivation. However, mechanisms of TORC1 regulation in fission yeast remain poorly understood. In this study, we found that Pef1, which is an ortholog of mammalian CDK5, regulates the initiation of sexual differentiation through positive regulation of TORC1 activity. Conversely, deletion of pef1 leads to activation of autophagy and subsequent excessive TORC1 reactivation during the early phases of the nitrogen starvation response. This excessive TORC1 reactivation results in the silencing of the Ste11-Mei2 pathway and mating defects. Additionally, we found that pef1 genetically interacts with tsc1 and tsc2 for TORC1 regulation, and physically interacts with three cyclins, Clg1, Pas1 and Psl1. The double deletion of clg1 and pas1 promotes activation of autophagy and TORC1 during nitrogen starvation, similar to what is seen in pef1Δ cells. Overall, our work suggests that Pef1–Clg1 and Pef1–Pas1 complexes regulate initiation of sexual differentiation through control of the TSC–TORC1 pathway and autophagy.

scholarly journals Autophagy-deficient Schizosaccharomyces pombe mutants undergo partial sporulation during nitrogen starvation

Microbiology ◽  
2009 ◽  
Vol 155 (12) ◽  
pp. 3816-3826 ◽  
Author(s):  
Hiroyuki Mukaiyama ◽  
Shiro Kajiwara ◽  
Akira Hosomi ◽  
Yuko Giga-Hama ◽  
Naotaka Tanaka ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nitrogen Source ◽  
Environmental Changes ◽  
Nitrogen Starvation ◽  
Nitrogen Sources ◽  
Vacuole Fusion ◽  
Sporulation Process

Autophagy is triggered when organisms sense radical environmental changes, including nutritional starvation. During autophagy, cytoplasmic components, including organelles, are enclosed within autophagosomes and are degraded upon lysosome–vacuole fusion. In this study, we show that processing of GFP-tagged Atg8 can serve as a marker for autophagy in the fission yeast Schizosaccharomyces pombe. Using this marker, 13 Atg homologues were also found to be required for autophagy in fission yeast. In budding yeast, autophagy-deficient mutants are known to be sterile, whereas in fission yeast we found that up to 30 % of autophagy-defective cells with amino acid auxotrophy were able to recover sporulation when an excess of required amino acids was supplied. Furthermore, we found that approximately 15 % of the autophagy-defective cells were also able to sporulate when a prototrophic strain was subjected to nitrogen starvation, which suggested that fission yeast may store sufficient intracellular nitrogen to allow partial sporulation under nitrogen-limiting conditions, although the majority of the nitrogen source is supplied by autophagy. Monitoring of the sporulation process revealed that the process was blocked non-specifically at various stages in the atg1Δ and atg12Δ mutants, possibly due to a shortage of amino acids. Taking advantage of this partial sporulation ability of fission yeast, we sought evidence for the existence of a recycling system for nitrogen sources during starvation.

Analysis of the Schizosaccharomyces pombe cyclin puc1: evidence for a role in cell cycle exit

1994 ◽  
Vol 107 (3) ◽  
pp. 601-613 ◽  
Author(s):  
S.L. Forsburg ◽  
P. Nurse
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Sexual Development ◽  
Mitotic Cycle ◽  
Nitrogen Starvation ◽  
Cell Cycle Exit ◽  
Gene Encoding ◽  
Yeast Saccharomyces Cerevisiae ◽  
Protein Blocks

The puc1+ gene, encoding a G1-type cyclin from the fission yeast Schizosaccharomyces pombe, was originally isolated by complementation in the budding yeast Saccharomyces cerevisiae. Here, we report the molecular characterization of this gene and analyse its role in S. pombe. We fail to identify any function of this cyclin at the mitotic G1/S transition in S. pombe, but demonstrate that it does function in exit from the mitotic cycle. Expression of the puc1+ gene is increased during nitrogen starvation, and puc1 affects the timing of sexual development in response to starvation. Overexpression of the puc1 protein blocks sexual development, and rescues pat1ts cells, which would otherwise undergo a lethal meiosis. We conclude that puc1 contributes to negative regulation of the timing of sexual development in fission yeast, and functions at the transition between cycling and non-cycling cells.

Pap1p-dependent upregulation of thioredoxin 3 and thioredoxin reductase genes from the fission yeast under nitrosative stress

2012 ◽  
Vol 58 (2) ◽  
pp. 206-211 ◽  
Author(s):  
Min-Sik Park ◽  
Hyeon-Jung Kim ◽  
A Rum Park ◽  
Kisup Ahn ◽  
Hye-Won Lim ◽  
...  
Keyword(s):  
Fission Yeast ◽  
Nitrosative Stress ◽  
Thioredoxin Reductase

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.

Sign in / Sign up

Export Citation Format

Share Document