• Title/Summary/Keyword: S. pombe

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Cloning and Sequence Analysis of the Kinesin Gene in Schizosaccharomyces pombe (Schizosaccharomyces pombe의 Kinesin 유전자의 클로닝과 염기서열분석)

  • 정재욱;최성민;김형배;이명석
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.18-24
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    • 1999
  • Kinesin has been discovered in Saccharomyces cerevisiae, Aspergillus nidulans, and Drosophila melanogaster and it has major roles in the movemenl of chromosomes and separation of spindle poles. In this study, a gene encoding kinesin heavy chain in Schizosaccharo~n)~ces pombe was cloned by using the polymerase chain reaction with degenerated primcrs corresponding to highly conserved regions of the kinesin heavy chain motor domain. The kinesin gene in S pombe contains an open reading frame of 2496 base pairs and encodes a kinesin prolein of 832 amino acids with a molecular weight of 96 kd. From thc comparison of the predictcd amino acids of the newly cloned kinesin, the kinesin in S. pornbe belongs to the C-terminal motor subfamily of kincsin-related protein.

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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein S20

  • Lee, Yoon-Jong;Kim, Kyunghoon;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Daemyung;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.31-36
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    • 2001
  • A cDNA clone encoding the ribosomal protein S20 has been isolated from the Schizosaccharomyces pombe cDNA library by colony hybridization. The insert contained in the original plasmid pYJ10 was transferred intro shuttle vector pRS316 generate plasmid pYJll. The dDNA insert of plasmid pYJll, contains 484 nucleotides and encodes a protein of 118 amino acids with a calculated mass of 13,544 daltons. The deduced amino acid sequence of S. pombe ribosomal protein S20 is very homologous with fruit fly, rat, and budding yeast counterparts. It is also homologous with Xenopus S22 ribosomal protein. S. pombe ribosomal protein S20 appears to be relatively hydruphobic except the C-terminal region. The 728 bp upstream region of the S20 gene was amplified from chromosomal DNA and transferred into the BamHI/EcoRI site of the promoterles $\beta$-galactosidase gene of the vector YEp357R, which resulted in fusion plasmid pYS20. The synthesis of $\beta$-galactosidase from the fusion plasmid appeared to be the highest in the mid-exponential phase. The S. pombe cells with the fusion plasmid grown at 35$\^{C}$ gave lower $\beta$-galactosidase activity than the cells grown at 30$\^{C}$. Computer analysis showed the consensus sequence CAGTCACA in the upstream regions of various ribosomal protein genes in S. pombe, which would be involved in the coordinated expression of small ribosomal proteins.

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Characterization of HRD3, a Schizosaccharomyces pombe Gene Involved in DNA Repair and Cell Viability

  • Choi, In-Soon
    • Animal cells and systems
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    • v.7 no.2
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    • pp.159-164
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    • 2003
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. The RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of a S. pombe RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (homolog of RAD3 gene). Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as HRD3 gene and this gene exists as a single copy in S. pombe. The transcript of 2.8 kb was detected by Northern blot analysis, The level of transcripts increased by ultraviolet (UV) irradiation, indicating that HRD3 is one of the UV-inducible genes in S. pombe. Furthermore, the predicted partial sequence of HRD3 protein has 60% identity to S. cerevisiae RAD3 gene. This homology was particularly striking in the regions identified as being conserved in a group of DNA helicases. Gene deletion experiments indicate that the HRD3 gene is essential for viability and DNA repair function. These observations suggest evolutionary conservation of other protein components with which HRD3 might interact in mediating its DNA repair and viability functions.

Schizosaccharomyces pombe의 연구 동향 - 효모의 연구동향

  • 김형배
    • The Microorganisms and Industry
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    • v.19 no.4
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    • pp.55-61
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    • 1993
  • 이 논문에서는 S. pombe의 특징과 연구방법에 대하여 전반적인 검토를 하고, human 등 고등생물의 연구에 대한 model system으로의 S. pombe에 대한 연구를 살펴보고자 한다. 특히 많은 연구분야중 현재 가장 활발히 연구되고 있는 분야인 cell cycle에 대한 연구와 oncogene에 대한 연구, 그리고 이를 이용한 차원높은 진핵 세포에서의 연구에 대하여 논하고자 한다.

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Participation of protein disulfide isomerase 2 in the tolerance against mercury toxicity in Schizosaccharomyces pombe (수은 독성에 대한 Schizosaccharomyces pombe 단백질2황화물이성질화효소 2의 저항성)

  • Choi, Jiye;Lim, Chang-Jin;Kim, Kyunghoon
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.338-346
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    • 2015
  • The present work was undertaken to address the role of protein disulfide isomerase 2 (Pdi2) in the mercury-tolerance of Schizosaccharomyces pombe, using the Pdi2-overexpressing recombinant plasmid pYPDI2 and the corresponding vector plasmid pRS316. When exposed to mercuric chloride, the PDI2 overepxression cells grew significantly better than the vector control cells. They revealed the lower levels of intracellular reactive oxygen species (ROS) and nitric oxide (NO), when incubated with mercuric chloride for 6 h, than the vector control cells. The PDI2 overepxression cells contained the higher levels of total glutathione (GSH) and superoxide dismutase (SOD) activity than the vector control cells, after 6 h of incubation in mercuric chloride. However, the PDI2 overepxression cells contained similar levels of glutathione peroxidase (GPx) activities, compared to those of the vector control cells. Taken together, the S. pombe Pdi2 promotes the tolerance against mercury toxicity through up-regulating total GSH and SOD and subsequently attenuating ROS and NO elevations.

Ribonucleotide Reductase Activity of Schizosaccbarornyces pombe Is Inhibited by Escherichia coli RecA Antibody (Schizosaccbarornyces pombe에서 Escherichia coli RecA 항체에 의한 Ribonucleotide Reductase 효소활성 저해)

  • Lee, Jung-Sup;Chun, Min-Suck;Kim, Ok-Bong;Park, Jong-Kun;Kim, Si-Wouk;Park, Yeal;Yang, Young-Ki;Hong, Seung-Hwan;Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.565-569
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    • 1995
  • We have previoosly demonstrated that the RecA-like protein of Schizosaccharomyces pombe (S. pombe) is immunologically related to Escherichia coil (E. coil) RecA protein and that the cellular level of the protein is significantly increased by inhibitors of nucleotide pool-forming enzymes such as hydroxyurea (HU) and methotrexate (MTX) (lee and Park, 1994; lee et al., 1994). In this study, we report that the ribonudeotide redudase activity of S. pombe is inhibited by E. coil RecA antibody, as determined by thin layer chromatography using [5-$^3$H]CDP as a substrate. The relative activity of ribonucleotide reductase was dramatically inhibited by 100 mM of flu (26.4% reduction) in in vitro assay, compared to that of non-treated control. The ribonucleotide reductase activity was also inhibited by immunoprecipitation with E. coil RecA antibody (43.3% reduction). These results indicate that the strudure of S. pombe ribonucleotide reductase is in part similar to that of E. coil RecA protein.

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Regulation of Thioltransferase Activity from Schizosaccharomyces pombe

  • Cho, Young-Wook;Park, Eun-Hee;Lim, Chang-Jin
    • BMB Reports
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    • v.33 no.5
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    • pp.422-425
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    • 2000
  • Thioltransferase (TTase), also known as glutaredoxin (Grx), is an enzyme catalyzing the reduction of a variety of disulfide compounds and acting as a cofactor for various enzymes such as ribonucleotide reductase. The Schizosaccharomyces pombe cells, exponentially grown in rich medium at $30^{\circ}C$, were shifted to $20^{\circ}C$ and $35^{\circ}C$. The yeast cells, shifted to $35^{\circ}C$, showed higher TTase activity than the cells continuously grown at $30^{\circ}C$, whereas the yeast cells, shifted to $20^{\circ}C$, gave lower TTase activity. The S. pombe cells, exponentially grown in minimal medium and shifted from $30^{\circ}C$ to $35^{\circ}C$ and $40^{\circ}C$, produced higher TTase activity. When the S. pombe cells were initially incubated in rich and minimal media at three different temperatures ($25^{\circ}C$, $30^{\circ}C$ and $35^{\circ}C$), they showed higher TTase activity at higher temperature. These results suggest that the TTase activity of S. pombe is regulated by temperature.

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Cloning and Expression of Alkaline Phosphatase Gene from Schizosaccharomyces pombe

  • Kang, Sung-Won;Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.3
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    • pp.262-267
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    • 2001
  • A cDNA coding alkaline phosphatase (AP) homologue was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The nucleotide sequence of the cloned cDNA appeared to lack the N-terminal coding region. The genomic DNA encoding alkaline phosphatase homologue was isolated from S. pombe chromosomal DNA using PCR. The amplified DNA fragment was ligated into plasmid pRS315 to generate the recombinant plasmid pSW20. The DNA insert was subcloned as two smaller fragments for nucleotide sequencing. The sequence contains 2,789 by and encodes a protein of 532 amino acids with a molecular mass of 58,666 daltons. The S. pombe cells containing plasmid pSW20 showed much higher AP activity compared with the yeast cells with vector only This indicates that the cloned AP gene apparently encodes AP The predicted amino acid sequence of the S. pombe AP shares homology with those of other known APs.

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Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.77-82
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    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

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Effects of fission yeast ortholog of THOC5 on growth and mRNA export in fission yeast (THOC5의 분열효모 이종상동체가 생장 및 mRNA export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.435-439
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    • 2015
  • THO/TREX complex plays an important role in transcriptional elongation, mRNA processing, nuclear RNA export, and genome stability. A fission yeast, Schizosaccharomyces pombe, SPBC577.04 gene encoding the ortholog of THOC5, a component of THO/TREX complex, was identified and characterized. The S. pombe thoc5 (spthoc5) is not essential for both growth and mRNA export, but deletion of the spthoc5 gene caused growth defect and slight accumulation of $poly(A)^+$ RNA in the nucleus. And the functional spThoc5-GFP protein is localized mainly in the nucleus. Co-immunoprecipitation analysis showed that the Hpr1(THOC1) protein, an evolutionally well-conserved component of THO/TREX complex, interacted with spThoc5 as well as Tho2(THOC2), another subunit of THO complex. These results suggest that S. pombe Thoc5 as a component of THO/TREX complex is also involved in mRNA export from the nucleus.