• Title/Summary/Keyword: Yeast transcription

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Correlation Analysis between Regulatory Sequence Motifs and Expression Profiles by Kernel CCA

  • Rhee, Je-Keun;Joung, Je-Gun;Chang, Jeong-Ho;Zhang, Byoung-Tak
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.63-68
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    • 2005
  • Transcription factors regulate gene expression by binding to gene upstream region. Each transcription factor has the specific binding site in promoter region. So the analysis of gene upstream sequence is necessary for understanding regulatory mechanism of genes, under a plausible idea that assumption that DNA sequence motif profiles are closely related to gene expression behaviors of the corresponding genes. Here, we present an effective approach to the analysis of the relation between gene expression profiles and gene upstream sequences on the basis of kernel canonical correlation analysis (kernel CCA). Kernel CCA is a useful method for finding relationships underlying between two different data sets. In the application to a yeast cell cycle data set, it is shown that gene upstream sequence profile is closely related to gene expression patterns in terms of canonical correlation scores. By the further analysis of the contributing values or weights of sequence motifs in the construction of a pair of sequence motif profiles and expression profiles, we show that the proposed method can identify significant DNA sequence motifs involved with some specific gene expression patterns, including some well known motifs and those putative, in the process of the yeast cell cycle.

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Determination of the MYB Motif Interacting with WD40 and Basic Helix Loop Helix Proteins

  • Kim, Ji-Hye;Kim, Bong-Gyu;Ahn, Joong-Hoon
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.67-70
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    • 2012
  • Plant MYB transcription factors regulate secondary metabolism, cellular morphogenesis, and plant hormone signaling pathway. MYB proteins in plants consist of two repeats of 50 amino acid residues, which are referred to as R2R3 and they interact with WD40 or basic helix loop helix (bHLH) proteins. Yeast two hybrid assay was determined whether rice MYB protein interacts with either OsTTG1, which contains a WD40 domain, or with OsGL3, which contains a bHLH domain. Among 30 OsMYB proteins, three interacted with OsTTG1 and five interacted with OsGL3. A series of MYB mutants were created to determine the MYB domain important for the interaction with OsTTG1 or OsGL3. By using the yeast two hybrid assay, we found that the R3 motif of OsMYB10 and the R2 motif of OsMYB16 were required for interaction with OsTTG1 and OsGL3 proteins, respectively.

The Expanding Significance of Inositol Polyphosphate Multikinase as a Signaling Hub

  • Kim, Eunha;Ahn, Hyoungjoon;Kim, Min Gyu;Lee, Haein;Kim, Seyun
    • Molecules and Cells
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    • v.40 no.5
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    • pp.315-321
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    • 2017
  • The inositol polyphosphates are a group of multifunctional signaling metabolites whose synthesis is catalyzed by a family of inositol kinases that are evolutionarily conserved from yeast to humans. Inositol polyphosphate multikinase (IPMK) was first identified as a subunit of the arginine-responsive transcription complex in budding yeast. In addition to its role in the production of inositol tetrakis- and pentakisphosphates ($IP_4$ and $IP_5$), IPMK also exhibits phosphatidylinositol 3-kinase (PI3-kinase) activity. Through its PI3-kinase activity, IPMK activates Akt/PKB and its downstream signaling pathways. IPMK also regulates several protein targets non-catalytically via protein-protein interactions. These non-catalytic targets include cytosolic signaling factors and transcription factors in the nucleus. In this review, we highlight the many known functions of mammalian IPMK in controlling cellular signaling networks and discuss future challenges related to clarifying the unknown roles IPMK plays in physiology and disease.

Genome-Wide Identification of Haploinsufficiency in Fission Yeast

  • Baek, Seung-Tae;Han, Sang-Jo;Nam, Mi-Young;Kim, Young-Dae;Kim, Li-La;Lee, Hyun-Jee;Heo, Kyung-Sun;Lee, Hye-Mi;Lee, Min-Ho;Park, Song-Kyu;Maeng, Pil-Jae;Park, Young-Woo;Lee, Sung-Hou
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1059-1063
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    • 2008
  • Abnormal phenotypes resulting from haploinsufficiency (HI) are due to the loss of one allele. Recent studies in budding yeast have shown that HI originates from insufficient protein levels or from a stoichiometric imbalance between subunits of protein complexes. In humans, however, HI often involves transcription factors. Therefore, the species differences in HI and the molecular mechanisms of species-specific HI remain under investigation. In this study, HI in fission yeast was systematically surveyed. HI in fission yeast affected genes related to signaling and to basic cellular processes, as observed in budding yeast. These results suggest that there are species differences in HI and that the HI that occurs in fission yeast is intermediate to HI in budding yeast and humans.

Characterization of Osh3, an Oxysterol-binding Protein, in Filamentous Growth of Saccharomyces cerevisiae and Candida albicans

  • Hur, Hyang-Sook;Ryu, Ji-Ho;Kim, Kwang-Hoon;Kim, Jin-Mi
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.523-529
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    • 2006
  • OSH3 is one of the seven yeast homologues of the oxysterol binding proteins (OSBPs) which have the major binding affinity to the oxysterols and function as regulator of cholesterol biosynthesis in mammals. Mutational analysis of OSH3 showed that OSH3 plays a regulatory role in the yeast-to-hyphal transition through its oxysterol-binding domain in Saccharomyces cerevisiae. The OSH3 gene was also identified in the pathogenic yeast Candida albicans. Deletion of OSH3 caused a defect in the filamentous growth, which is the major cause of the C. albicans pathogencity. The filamentation defect of the mutation in the MAPK-associated transcription factor, namely $cph1{\Delta}$ was suppressed by overexpression of OSH3. These findings suggest the regulatory roles of OSH3 in the yeast filamentous growth and the functional conservations of OSH3 in S. cerevisiae and C. albicans.

효모시스템에서 Human Transglutaminase C(TGase II)의 발현에 관한 연구 : C-말단부위의 결손효과

  • Woo, Sang-Kyu;Jung, Sun-Mi;Rhee, Sang-Ki;Ahn, Byeong-Yoon;Kim, Hee-Chul
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.290-298
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    • 1996
  • In an effort to understand the role of the conserved domain and of the heterologous one-third part of the carboxy terminal domain of transglutaminase C (TGase II), attempts were made to express TGase II cDNA of human origin in yeast Saccharomyces cerevisiae as in a full-length form as well as in a form of C-terminal truncation. The 2$\mu$-based expression plasmids which contained the TGase II cDNA under the gal inducible promoter were introduced into yeast and the maintenance of the full-length and truncated form of the TGase II gene plasmids were confirmed by Southern blot. The expression of the TGase II gene was analysed by reverse transcription polymerase chain reaction (RT-PCR), and western blot analyses. As assayed by [1,4$^{14}$C]-putrescine incorporation into succinylated casein, the full-lenth as well as the truncated forms of recombinant TGase II showed some catalytic activity. These results indicate that the N-terminal homologous domain of human TGase II retains a catalytically active domain. The level of TGase II expressed in yeast, however, was far lower than satisfactory and other expression system should be sought further chracterization of the enzyme. The negative effect of TGase II on the growth of yeast is interesting with respect to the physiological effect of TGase II in cornification of epidermal keratinocytes.

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Synthesis and Secretion of the Endo-$\beta$-l,4-Glucanase from Bacillus subtilis in Industrial Yeast Strain (산업용 효모에서 Bacillus subtilis Endo-$\beta$-1,4-Glucanase의 생합성 및 분비)

  • 박용준;이영호;백운화;강현삼
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.348-355
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    • 1991
  • DNA segment encoding $\beta$-1, 4-glucanase of Bacillus subtilis was fused in frame to mouse $\alpha$-amylase signal sequence behind the alcohol dehydrogenase isoenzyme I gene (ADHI) promoter of the yeast expression vector pMS12. To enhance the expression level of the $\beta$glucanase gene in yeast, transcription terminator sequence iso-1-cytochrome c gene (CYCI) was inserted into the recombinant plasmid. The transformants harbouring such recombinant plasmids secreted $\beta$-glucanase into the culture medium. The expresstion level of the $\beta$-glucanase gene was increased about 2-fold caused by inserting the terminator. The amount of the secreted $\beta$-glucanase in culture medium was approximately 60% of the total quantity synthesized.

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Cloning and Characterization of the Orotidine-5'-Phosphate Decarboxylase Gene (URA3) from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.642-648
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    • 2012
  • We determined the nucleotide sequence of the URA3 gene encoding orotidine-5'-phosphate decarboxylase (OMPDCase) of the erythritol-producing osmotolerant yeast Candida magnoliae by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 795 bp, encoding a 264 amino acid residue protein with the highest identity to the OMPDCase of the yeast Kluyveromyces marxianus. Phylogenetic analysis of the deduced amino acid sequence revealed that it shared a high degree of identity with other yeast OMPDCase homologs. The cloned URA3 gene successfully complemented the ura3 null mutation in Saccharomyces cerevisiae, revealing that it encodes a functional OMPDCase in C. magnoliae. An enzyme activity assay and reverse transcription polymerase chain reaction indicated that the expression level of the C. magnoliae URA3 gene in S. cerevisiae was not as high as that of the S. cerevisiae URA3 gene. The GenBank accession number for C. magnoliae URA3 is JF521441.

Forkhead Genes are Key Regulators of Developmental Processes in Aspergillus nidulans

  • Oh, Dong-Soon;Kim, Jong-Hwa;Han, Dong-Min;Han, Kap-Hoon
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.39-39
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    • 2014
  • In a homothallic filamentous fungus Aspergillus nidulans, sexual and asexual developments are largely affected by the genetic and environmental factors. To regulate the complex subsets of genes involved in the developmental processes accurately, tight regulations of transcription factors are required. The forkhead type transcription factors are the class of regulators that function in a broad spectrum of cellular and developmental processes in many species from yeast to human. Here, we identified the fkhA and fkhB genes that encode a conserved forkhead transcription factors. The fkhA deletion resulted in the complete loss of fruiting body formation under all conditions favoring sexual development, suggesting that the fkhA gene is required for sexual development in A. nidulans. Overexpression of fkhA resulted in enhanced formation of fruiting bodies under induction condition not only in the normal condition but also in the condition of presence of 0.6 M KCl, which strongly inhibits sexual development. To know the function of the fkhB gene, we also generated fkhB knock-out strain in A. nidulans. Deletion of fkhB resulted in abnormal conidiophore formation under standard conditions and delayed sexual development process, suggesting that the fkhB gene plays an important role in conidiophore morphogenesis Taken together, these results suggest that the fkhA gene is necessary and sufficient for regulating sexual development and the fkhB gene is a transcription factor related in asexual developmental process in A. nidulans.

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Potential role of the histone chaperone, CAF-1, in transcription

  • Kim, Hye-Jin;Seol, Ja-Hwan;Cho, Eun-Jung
    • BMB Reports
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    • v.42 no.4
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    • pp.227-231
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    • 2009
  • The eukaryotic genome forms a chromatin structure that contains repeating nucleosome structures. Nucleosome packaging is regulated by chromatin remodeling factors such as histone chaperones. The Saccharomyces cerevisiae H3/H4 histone chaperones, CAF-1 and Asf1, regulate DNA replication and chromatin assembly. CAF-1 function is largely restricted to non-transcriptional processes in heterochromatin, whereas Asf1 regulates transcription together with another H3/H4 chaperone, HIR. This study examined the role of the yeast H3/H4 histone chaperones, Asf1, HIR, and CAF-1 in chromatin dynamics during transcription. Unexpectedly, CAF-1 was recruited to the actively transcribed region in a similar way to HIR and Asf1. In addition, the three histone chaperones genetically interacted with Set2-dependent H3 K36 methylation. Similar to histone chaperones, Set2 was required for tolerance to excess histone H3 but not to excess H2A, suggesting that CAF-1, Asf1, HIR, and Set2 function in a related pathway and target chromatin during transcription.