• Title/Summary/Keyword: Schizosaccharomyces pombe

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Immunofluorescence Localization of Schizosaccharomjyces pombe $cdc103^{+}$ Gene Product

  • Kim, Hyong-Bai
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.248-254
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    • 1996
  • $cdc103^+$ gene in Schizosaccharomyces pombe which is similar to the CDC3 gene in Saccharomyces cerevisiae was cloned and sequenced. Comparison of the predicted amino acid sequences of $cdc103^+$ and CDC3 revealed that they share significant similarity (43% identity and 56% identity or similarity) to each other. The gene product of CDC3 in S. cerevisiae is known to be a highly ordered ring of filaments that lies just inside the cytoplasmic membrane in the region of the mother-bud neck. In order to characterize the gene product of $cdc103^+$ in Schizosaccharomyces pombe, fusion proteins were used to generate the polyclonal antibodies specific for the gene product (cdc103p). In immunofluorescence experiments, these antibodies decorate the region of the septum formation as a double ring structure late in the cell division cycle.

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Expression, Characterization and Regulation of a Saccharomyces cerevisiae Monothiol Glutaredoxin (Grx6) Gene in Schizosaccharomyces pombe

  • Lee, Jae-Hoon;Kim, Kyunghoon;Park, Eun-Hee;Ahn, Kisup;Lim, Chang-Jin
    • Molecules and Cells
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    • v.24 no.3
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    • pp.316-322
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    • 2007
  • Glutaredoxins (Grxs), also known as thioltransferases (TTases), are thiol oxidoreductases that regulate cellular redox state in a variety of organisms. In the budding yeast Saccharomyces cerevisiae, Grx1 and 2 are cytosolic dithiol Grxs, while Grx3, 4 and 5 are monothiol Grxs. A gene encoding a new monothiol Grx, Grx6, was cloned from the genomic DNA of S. cerevisiae by PCR. Its DNA sequence contains 1,080 bp, and encodes a putative protein of 203 amino acid residues containing Cys-Phe-Tyr-Ser at the active site. Grx6 is similar to other monothiol Grxs in the same organism and to Grx3 in the fission yeast Schizosaccharomyces pombe. and its predicted three-dimensional structure resembles that of S. pombe Grx3. S. pombe cells harboring plasmid pFGRX6 containing the Grx6 gene had about 1.3-fold elevated Grx activity in the exponential phase, and grew better than the control cells under some stressful conditions. Synthesis of ${\beta}$-galactosidase from a Grx6-lacZ fusion gene in S. pombe was enhanced by potassium chloride, aluminum chloride and heat ($37^{\circ}C$) treatment. S. pombe cells harboring plasmid pFGRX6 had elevated ROS levels whereas S. pombe cells harboring extra copies of Grx3 had reduced ROS levels.

Functional Characterization of the Madlp, a Spindle Checkpoint Protein in Fission Yeast

  • Kim, In-Gyu;Rhee, Dong-Keun;Lee, Hee-Cheul;Lee, Joo;Kim, Hyong-Bai
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.694-700
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    • 2005
  • Defects in the mitotic spindle or in the attachment of chromosomes to the spindle are believed to release an activated form of spindle checkpoint complex that inhibits APC-dependent ubiquitination and subsequently arrests the cell cycle at metaphase. When the spindle assembly is disrupted, the fission yeast mitotic arrest deficient (mad) mutants fail to arrest and rapidly lose viability. To enhance our understanding of the molecular mechanisms for the pathway of checkpoint function, the functional characterizations of Mad 1 p from Schizosaccharomyces pombe involved in this process have been carried out. Yeast two-hybrid and various deletion analyses of S. pombe Mad1 p reveal that the C terminus of Mad1p is critical for the binding of Mad2p and maintenance of Mad 1 p-Mad2p interaction. In addition, it was found. that the Mad1p region (residues 206-356) is essential for Mad1p-other checkpoint components. Mad1p truncating this region is sufficient to bind Mad2p but abolishes the checkpoint function, indicating that the checkpoint function is necessary for interaction of Mad 1 p-other checkpoint components. The possible functions of S. pombe Mad1p at the cell cycle checkpoint are discussed.

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.

Effects of Tho2, a component of THO complex, on growth and mRNA export in fission yeast (분열효모에서 THO 복합체의 구성요소인 Tho2가 생장 및 mRNA export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.181-185
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    • 2015
  • Tho2/THOC2 is a subunit of the THO complex that plays important roles in mRNP biogenesis connecting transcription with mRNA maturation and export. A fission yeast, Schizosaccharomyces pombe, ortholog of Tho2/THOC2 was identified from the genome database. Tetrad analysis showed that the S. pombe tho2 is essential for growth. Repression or overexpression of the tho2 gene caused growth defect with elongated cells, abnormal DNA distribution, and accumulation of $poly(A)^+$ RNA in the nucleus. And the functional GFP-Tho2 protein is localized mainly in the nucleus. Yeast two-hybrid analysis showed that Tho2 interacted with Tex1, another subunit of THO complex. These results suggest that S. pombe Tho2 is also involved in mRNA export from the nucleus and is a component of THO complex.

Effects of Cdc31, a component of TREX-2 complex, on growth and mRNA export in fission yeast (분열효모에서 TREX-2 복합체의 구성요소인 Cdc31이 생장과 mRNA export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.383-387
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    • 2016
  • In fission yeast, Schizosaccharomyces pombe, the cdc31 gene encodes a member of the conserved $Ca^{2+}$-binding centrin/CDC31 family, which is a component of spindle pole body. Here, we demonstrate that the S. pombe cdc31p is also a component of TREX-2 complex, which influences mRNA export from the nucleus to the cytoplasm. Repression of the cdc31 gene expression caused growth defect with accumulation of $poly(A)^+$ RNA in the nucleus. On the other hand, over-expression of cdc31 exhibited no defects of both growth and bulk mRNA export, but showed somewhat longer cell morphology. Yeast two-hybrid analysis showed that Cdc31 interacted with Sac3 and Pci2, the subunits of TREX-2 complex. These results suggest that S. pombe Cdc31 is also involved in mRNA export as a component of TREX-2 complex.

효모에 의한 과실주중의 감산 효과에 관한 연구 제1보 균주의 분리 및 동정

  • 유대식
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1978.04a
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    • pp.96.1-96
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    • 1978
  • 미생물학적 감산현상에 의하여 과실주중의 산미를 조절하기 위한 방법으로 효모에 의하여 사과산을 알코올로 분해하는 Maloalcohol 발효를 유도하고자 하여 본 연구를 하였다. 이미 발표된 Schizosaccharomyces pombe 0-77보다 단 시간에 강력히 사과산을 소비하는 효모균을 딸기의 과피로부터 분리, 동정하였다. 본 분리균을 J. Lodder의 “The Yeasts”에 준하여 동정한 바 분열법에 의하여 증식하며 포자를 형성하고 galactose 를 발효못하므로 Schizosacoha-romyces 속으로 분류하였다. 위균사를 잘 형성하며 melibiose를 발효하므로 Schizosaccharomyces japonicus와 일치 하였다. 그러나 변종의 동정은 어려우나 위균사의 형태로 보아 Schizosaccharomyces japonicus var. japonicus로 동정하였다.

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Transcriptional Regulation of the Gene Encoding ${\gamma}$-Glutamylcysteine Synthetase from the Fission Yeast Schizosaccharomyces pombe

  • Kim, Su-Jung;Kim, Hong-Gyum;Kim, Byung-Chul;Kim, Kyunghoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.233-238
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    • 2004
  • Transcriptional regulation of the Schizosaccharomyces pombe y-glutamylcysteine synthetase (GCS) gene was examined using the two GCS-lacZ fusion plasmids pUGCS101 and pUGCS102, which harbor 607 bp and 447 bp upstream regions, respectively. The negatively-acting sequence was located in the -607 - -447 bp upstream region of the GCS gene. The upstream sequence responsible for induction by menadione(MD) and L-buthionine-(S, R)-sulfoximine (BSO) resides in the -607 - -447 bp region, whereas the sequence which codes for nitric oxide induction is located within the -447 bp region, measured from the translational initiation point. Carbon source-dependent regulation of the GCS gene appeared to be dependent on the nucleotide sequence within -447 bp region. The transcription factor Papl is involved in the induction of the GCS gene by MD and BSO, but not by nitric oxide. Induction of the GCS gene occurring due to low glucose concentration does not depend on the presence of Pap1. These data imply that induction by MD and BSO may be mediated by the Pap1 binding site, probably located in the -607 - -447 region, and also that the nitric oxide-mediated regulation of the S. pombe GCS gene may share a similar mechanism with its carbon-dependent induction.

Purification and Characterization of Hrp1, a Homolog of Mouse CHD1 from the Fission Yeast Schizosaccharomyces pombe

  • Yong Hwan Jin;Eung Jae Yoo;Yeun Kyu Jang;Seung Hae Kim;Chee-Gun Lee;Rho Hyun Seong;Seung Hwan Hong;Sang Dai Park
    • Animal cells and systems
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    • v.2 no.4
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    • pp.539-543
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    • 1998
  • Hrp1, of Schizosaccharomyces pombe, is a new member of the SW12/SNF2 protein family that contains a chromodomain and a DNA binding domain as well as ATPase/7 helicase domains. This configuration suggests that Hrp1 could be a homolog of mouse CHD1, which is thought to function in altering the chromatin structure to facilitate gene expression. To understand the enzymatic nature of Hrp1 we purified the 6-Histidine-tagged Hrp1 protein (6$\times$His-Hrp1) to homogeneity from a S. pombe Hrp1-overexpressing strain and hen examined its biochemical properties. We demonstrate that the purified 6$\times$His-Hrp1 protein exhibited a DNA-binding activity with a moderate preference to the (A+T)-rich tract in double-stranded NA via a minor groove interaction. However, we failed to detect any intrinsic DNA helicase activity from the purified Hrp1 like other SW12/SNF2 proteins. These observations suggest that the DNA binding activities of Hrp1 may be involved in the remodeling of the chromatin structure with DNA-dependent ATPase. We propose that Hrp1 may function in heterochromatins as other proteins with a chromo- or ATPase/helicase domain and play an important role in the determination of chromatin architecture.

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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein L11

  • Kim, Hong-Gyum;Lee, Jin-Joo;Park, Eun-Hee;Sa, Jae-Hoon;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.4
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    • pp.379-384
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    • 2001
  • The cDNA encoding ribosomal protein was identified from a cDNA library of Schizosaccharomyces pombe. The nucleotide sequence of the 548 by cDNA clone reveals an open reading frame, which encodes a putative protein of 166 amino acids with a molecular mass of 18.3 kDa. The amino acid sequence of the S. pombe L11 protein is highly homologous with those of rat and fruit, while it is clearly less similar to those of prokaryotic counterparts. The 1,044 by upstream sequence, and the region encoding N-terminal 7 amino acids of the genomic DNA were fused into the promoterless $\beta$-galactosidase gene of the shuttle vector YEp357 in order to generate the fusion plasmid pHY L11. Synthesis of $\beta$-galactosidase from the fusion plasmid varied according to the growth curve. It decreased significantly in the growth-arrested yeast cells that were treated with aluminum chloride and mercuric chloride. However, it was enhanced by treatments with cadmium chloride ($2.5\;{\mu}M$), zinc chloride ($2.5\;{\mu}M$), and hydrogen peroxide (0.5 mM). This indicates that the expression of the L,11 gene could be induced by oxidative stress.

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