• Title/Summary/Keyword: fission yeast

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Construction of Schizosaccharomyces pombe spThp1 Null Mutants and its Characterization (분열효모 Schizosaccharomyces pombe에서 spThp1 유전자 결실돌연변이의 제조와 특성 조사)

  • Yoon Jin-Ho
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.149-152
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    • 2006
  • The sp%pl null mutant was constructed to study the function of fission yeast Schizosaccharomyces pombe spThp1, which is homologous to budding yeast Saccharomyces cerevisiae THP1. Tetrad analysis showed that the spThp1 is not essential for vegetative growth. The spThp1 null mutant also showed no massive poly(A)+ RNA export defect. However, spThp1 null is genetically associated with spMex67 null. These results suggest that spThp1 is involved in mRNA export out of the nucleus.

Construction of spSac3 Null Mutants Defective in mRNA Export (mRNA의 핵에서 세포질로의 이동에 관여하는 spSac3 유전자의 결실돌연변이 제조와 특성 조사)

  • Kang Sook-Hee;Yoon Jin-Ho
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.153-155
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    • 2006
  • We constructed the null mutants of fission yeast Schizosaccharomyces pombe spSac3 gene that is homologous to budding yeast Saccharomyces cerevisiae SAC3 involved in mRNA export out of nucleus. Tetrad analysis showed that the spSac3 is essential for vegetative growth. The spSac3 mutants harboring pREP81X-spSac3 plasmid showed poly(A)+ RNA export defect in the presence of thiamine. These results suggest that spSac3 is also involved in mRNA export from the nucleus.

Fission Yeast-based Screening to Identify Putative HDAC Inhibitors Using a Telomeric Reporter Strain

  • Chung, Kyung-Sook;Ahn, Jiwon;Choi, Chung-Hae;Yim, Nam Hui;Kang, Chang-Mo;Kim, Chun-Ho;Lee, Kyeong;Park, Hee-Moon;Song, Kyung-Bin;Won, Misun
    • Molecules and Cells
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    • v.26 no.1
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    • pp.93-99
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    • 2008
  • Transcriptional silencing is regulated by promoter methylation and histone modifications such as methylation and acetylation. We constructed a Schizosaccaromyces pombe reporter strain, KCT120a, to identify modifiers of transcriptional silencing, by inserting the $ura4^+$ gene into a heterochromatic telomere region. Two compounds inhibited the activity of histone deacetylases, induced acetylation of histone H3 and caused apoptotic cell death in HeLa cells. Expression of gelsolin and $p21^{waf1/cip1}$ also increased, as it does in response to HDAC inhibitors such as TSA. Therefore, these compounds appear to be potent inhibitors of HDACs, and hence potential anti-cancer drugs. Our observations suggest that a yeast cell-based assay system for transcriptional silencing may be useful for identifying histone deacetylase inhibitors and other agents affecting chromatin remodeling.

A Second Thioltransferase of Schizosaccharomyces pombe Contains Glutathione S-transferase Activity

  • Kim, Hong-Gyum;Park, Eun-Hee;Lim, Chang-Jin
    • BMB Reports
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    • v.32 no.6
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    • pp.535-540
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    • 1999
  • Two types of the thioltransferase (also called glutaredoxin) have been previously detected in the cytosolic extract of Schizosaccharomyces pombe, a fission yeast. Previously, the one with a smaller molecular mass (14kDa) was purified and characterized. In the present study, the second thioltransferase was purified. The purification procedure included ammonium sulfate fractionation (40-80%), Sephadex G-200 gel filtration, DEAE-cellulose ion-exchange chromatography, Sephadex G-50 gel filtration, and glutathione-agarose affinity chromatography. The purified enzyme showed a single band on SDS-PAGE, and its molecular mass was determined to be 23 kDa. It utilizes various compounds as substrates, including 2-hydroxyethyl disulfide. Interestingly, we found that the purified thioltransferase also contains significant glutathione S-transferase activity.

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The Heterochromatin Protein 1 (HP1) Family: Put Away a Bias toward HP1

  • Kwon, So Hee;Workman, Jerry L.
    • Molecules and Cells
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    • v.26 no.3
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    • pp.217-227
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    • 2008
  • Heterochromatin protein 1 (HP1) was first described in Drosophila melanogaster as a heterochromatin associated protein with dose-dependent effect on gene silencing. The HP1 family is evolutionarily highly conserved and there are multiple members within the same species. The multi-functionality of HP1 reflects its ability to interact with diverse nuclear proteins, ranging from histones and transcriptional co-repressors to cohesion and DNA replication factors. As its name suggests, HP1 is well-known as a silencing protein found at pericentromeres and telomeres. In contrast to previous views that heterochromatin is transcriptionally inactive; noncoding RNAs transcribed from heterochromatic DNA repeats regulates the assembly and function of heterochromatin ranging from fission yeast to animals. Moreover, more recent progress has shed light on the paradoxical properties of HP1 in the nucleus and has revealed, unexpectedly, its existence in the euchromatin. Therefore, HP1 proteins might participate in both transcription repression in heterochromatin and euchromatin.

Possible Roles of LAMMER Kinase Lkh1 in Fission Yeast by Comparative Proteome Analysis

  • Cho, Soo-Jin;Kim, Young-Hwan;Park, Hee-Moon;Shin, Kwang-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.108-112
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    • 2010
  • To investigate the possible roles of LAMMER kinase homologue, Lkh1, in Schizosaccharomyces pombe, whole proteins were extracted from wild type and lkh1-deletion mutant cells and subjected to polyacrylamide gel electrophoresis. Differentially expressed proteins were identified by tandem mass spectrometry (MS/MS) and were compared with a protein database. In whole-cell extracts, 10 proteins were up-regulated and 9 proteins were down-regulated in the mutant. In extracellular preparations, 6 proteins were up-regulated in the lkh1+ null mutant and 4 proteins successfully identified: glycolipid anchored surface precursor, $\beta$-glucosidase (Psu1), cell surface protein, glucan 1,3-$\beta$-glucosidase (Bgl2), and exo-1,3 $\beta$-glucanase (Exg1). These results suggest that Lkh1 is involved in regulating cell wall assembly.

Analysis of a Region Required for the Functions of Fission Yeast Nucleoporin Nup184 and Its SUMO Modification (분열효모 핵공단백질인 Nup184의 기능에 필요한 부위 분석 및 SUMO 변성)

  • Chai, Ai-Ree;Jang, Soo-Yeon;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.66-72
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    • 2012
  • The Nup188 protein is one of the largest evolutionally conserved nucleoprins (Nups) that compose the inner ring of nuclear pore complex (NPC). The Nup184 protein, fission yeast Schizosaccharomyces pombe ortholog of Nup188p, is required for normal growth and mRNA export in nutrient-rich medium (YES). Here, we identified a carboxyl region (482 to 1628) of Nup184 protein that was enough to complement the defects of both growth and mRNA export when the ${\Delta}nup184$ knock-out mutant was grown in YES medium. This region is also required for localization of GFP-Nup184 fusion to the nuclear periphery. In addition, we found that ORF of Nup184 (predicted 1564 amino-acid protein) registered in S. pombe GeneDB (hosted by Sanger Institute, UK) is 64 amino-acid residues shorter than that predicted by our sequence data. This carboxy-terminal region is necessary for the functions of Nup184p. We further demonstrated that Nup184 protein was conjugated with SUMO in vivo.

Studies on Intracellular Functions of the mas3 Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서 mas3 유전자의 세포내 기능 연구)

  • Hwang Mi Ra;Cha Jae Young;Shin Sang Min;Park Jong Kun
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.124-131
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    • 2005
  • The regulation of gene expression plays an important rolet in cell cycle controls. In this study, a novel $mas3^+$ (mitosis associated protein) gene, a homolog of human SMARCADl, was isolated and characterized from a fission yeast Schizosaccharomyces pombe. The overall homology between the helicase proteins of the two species is $87\%$. This DEAD/H box-containing molecule has seven highly conserved sequence regions that allow us to place it in the SNF2 family of the helicase superfamily. Knock-out cell of $mas3^+$ gene was constructed using kanMX6 as a selection marker. Survival of mas3 null mutant exposed to UV or MMS was similar to those of wild type cells. $mas3^+$ expression was lowest at $G_2$ and gradually increased. Cytokinesis of mas3 null mutant was abnormal at $26^{\circ}C\;and\;35^{\circ}C$ and a large number of multi-septate cells were produced. These results indicate that the $mas3^+$ is involved in cytokinesis and cell shape control.