• Title/Summary/Keyword: RNA polymerase II

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Two new species of Trichoderma isolated from commercially grown oyster mushroom, Pleurotus ostreatus (oral)

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.127.1-127
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    • 2003
  • We describe two new Trichoderma species associated with oyster mushroom in Korea. Trichoderma green mould has been one of the most serious diseases of oyster mushroom in Korea. Of these the predominant species are two unrecorded species. We designed as Trichoderma sp. Korean type 1 (Th K1) and Trichoderma sp. Korean type 2 (Th K2), respectively. Th K1 and Th K2 can be distinguished from previously reported Trichoderma species as well as each other in morphological characteristics including growth rate at 35$^{\circ}C$, colony morphology, conidia shape and branch pattern of phialides. Sequence of the ITS region of rDNA, the protein coding translation elongation factor gene(EF-1${\alpha}$), and RNA polymeraseII (RPB2) not only clearly separated Trichoderma sp. Korean types from their closely related T. harzianum biotype but also distinguished them from each other. Analyses of the EF-1${\alpha}$ and RPB2 sequences were found to be more useful for establishing systematic relationships among Trichoderma isolates than those of the ITS sequence. Based on the results of morphological and molecular characteristics. We propose the two Trichoderma sp. Korean types as the new species

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Functional Analysis of the Heptasequence SPTSPTY in the Transcriptional Activation Domain of Rat Nuclear Factor 1-A

  • Hwang, Jung-Su;Son, Kyung-No;Rho, Hyune-Mo;Kim, Ji-Young
    • BMB Reports
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    • v.32 no.5
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    • pp.468-473
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    • 1999
  • Nuclear Factor 1 (NF1) proteins are a family of transcriptional factors consisting of four different types: NF1-A, -B, -C, and -X. Some NF1 transcription factors contain a heptasequence motif, SPTSPSY, which is found as a repeat sequence in the carboxy terminal domain (CTD) of the largest subunit of RNA polymerase II. A similar heptasequence, SPTSPTY, is contained in rat liver NF1-A at a position between residues 469 and 475. In order to investigate the roles of the individual amino acids of the heptasequence of rat liver NF1-A in transcriptional activation, we systematically substituted single and multiple amino acid residues with alanine residue(s) and evaluated the transcriptional activities of the mutated NF1-A. Substitution of a single amino acid reduced transcriptional activity by 10 to 30%, except for the proline residue at position 473, whose substitution with alanine did not affect transcriptional activity. However, changes of all four serine and threonine residues to alanine or of the tyrosine residue along with the serine residue at position 469 to alanine reduced the activity to almost background levels. Our results indicate that multiple serine and threonine residues, rather than a single residue, may be involved in the modulation of the transcriptional activities of the factor. Involvement of the tyrosine residue is also implicated.

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Immunomodulating Activity of Fungal ${\beta}-Glucan$ through Dectin-1 and Toll-like Receptor on Murine Macrophage

  • Kim, Ha-Won
    • 한국약용작물학회:학술대회논문집
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    • 2006.11a
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    • pp.103-115
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    • 2006
  • [ ${\beta}-Glucan$ ] is a glucose polymer that has linkage of ${\beta}-(1,3)$, -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, ${\beta}-glucans$ are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding ${\beta}-glucans$ as pathogen-associated molecular pattern (PAMP). Recently ${\beta}-glucans$ receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-l is consisted of 7 exons and 6 introns. The polypeptide of dectin-l has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-l could recognize variety of beta-l,3 and/or beta-l,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-l mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-l was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with ${\beta}-glucans$ of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and $TNF-{\alpha}$ in the presence of LPS. However, GLG alone did not increase IL-6 nor $TNF-{\alpha}$ These results suggest that receptor dectin-l cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

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In vivo putative O-GlcNAcylation of human SCP1 and evidence for possible role of its N-terminal disordered structure

  • Koo, JaeHyung;Bahk, Young Yil
    • BMB Reports
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    • v.47 no.10
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    • pp.593-598
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    • 2014
  • RNA polymerase II carboxyl-terminal domain (RNAPII CTD) phosphatases are responsible for the dephosphorylation of the C-terminal domain of the small subunit of RNAPII in eukaryotes. Recently, we demonstrated the identification of several interacting partners with human small CTD phosphatase1 (hSCP1) and the substrate specificity to delineate an appearance of the dephosphorylation catalyzed by SCP1. In this study, using the established cells for inducibly expressing hSCP1 proteins, we monitored the modification of ${\beta}$-O-linked N-acetylglucosamine (O-GlcNAc). O-GlcNAcylation is one of the most common post-translational modifications (PTMs). To gain insight into the PTM of hSCP1, we used the Western blot, immunoprecipitation, succinylayed wheat germ agglutinin-precipitation, liquid chromatography-mass spectrometry analyses, and site-directed mutagenesis and identified the $Ser^{41}$ residue of hSCP1 as the O-GlcNAc modification site. These results suggest that hSCP1 may be an O-GlcNAcylated protein in vivo, and its N-terminus may function a possible role in the PTM, providing a scaffold for binding the protein(s).

Current Progress in the Analysis of Transcriptional Regulation in the Industrially Valuable Microorganism Aspergillus oryzae

  • Nakajima, Keiichi;Sano, Motoaki;Machida, Masayuki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.253-262
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    • 2000
  • Aspergillus is considered to be an attractive host for heterologous protein production because of its safety and ability to secrete large amounts of proteins. In order to obtain high productivity, thus far promoters of amylases have been most widely used in A. oryzae. Recent progress in cloning and expression analysis, including EST sequencing, revealed that glycolytic genes represent some of those most strongly expressed in A. oryzae. Therefore, promoters of glycolytic genes could be important alternatives to promoters of amylases because lower amounts of proteases are produced in the presence of glucose. Several A. oryzae transcription factors responsible for the induction and/or maximum expression of many industrially important genes encoding amylases and proteases have been cloned and characterized. In addition to the transcriptional regulatory factors, the gene encoding the largest subunit of RNa polymerase II, constituting the basic transcription machinery, has also been cloned from A. oryzae. This recently acquired understanding of the details of transcriptional regulatory mechanisms and factors will facilitate engineering flexible controls for the expression of proteins important for the fermentation industries.

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Molecular Cloning of the Gene Coding for 3-Isopropylmalate Dehydrogenase of Kluyveromyces fragilis (Kluyveromyces fragilis의 LEU gene의 Cloning)

  • 박성희;이동선;우주형;김종국;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.305-308
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    • 1990
  • In order to clone the gene coding for 3-isopropylmalate dehydrogenase of Muyveromyces fragilis, a shuttle plasmid vector pHNll4 was used. It can serve as a cloning vector in Saccharomyces cerevisiae DBY746 for other Sau3AI-cleaved DNA segment of Kluyveromyces fragilis. Two cloned fragments which complement the leu2 mutation of Saccharomyces cerevisiae and E, coli were obtained. Their length was 4.4 kb an 3.5 kb, and their orientation was opposite each other. From the fact that the two recombinant plasmids were expressed in Saccharomyces cerevisiae and E, coli, probably the two inserts had the promoter of Ktuyveromyces fi-agilis and that of Kluyveromyces fiagilis was efficiently assosiated with RNA polymerase of Saccharomyces cerevisiae and E. coli. According to the result of Southern hybridization, we thought that the cloned fragment has low homology with 3-isopropylmalate dehydrogenase coding region of E. coli and Saccharomyces cerevisiae.

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Re-evaluation of Hypocrea pseudogelatinosa and H. pseudostraminea isolated from shiitake mushroom (Lentinula edodes) cultivation in Korea and Japan

  • Kim, Chang Sun;Yu, Seung Hun;Nakagiri, Akira;Shirouzu, Takashi;Sotome, Kozue;Kim, Seon Cheol;Maekawa, Nitaro
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.341-356
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    • 2012
  • Shiitake (Lentinula edodes) is the most economically important cultivated mushroom, but yields are impacted by its competitor, Trichoderma spp. We previously found two unidentified Trichoderma species growing in bedlogs and sawdust shiitake media in Korea. Here, we identify and re-describe those two species based on molecular sequence data, morphology, and culture characteristics. Well-supported clades based on phylogenetic analyses of internal transcribed spacer, translation elongation factor 1-${\alpha}$, and RNA polymerase subunit II sequences grouped one of the unidentified Trichoderma spp. with Hypocrea pseudogelatinosa and the other with Hypocrea pseudostraminea, and their morphologies matched well with the original descriptions of the two Hypocrea species. This study reports the first phylogenetic analyses of H. pseudogelatinosa and Japanese strains of H. pseudostraminea. Based on the phylogenetic results, we redescribed these two species using modern taxonomic concepts in Hypocrea/Trichoderma.

Development of a Virus Elution and Concentration Procedure for Detecting Norovirus in Oysters

  • Ha, Sook-Hee;Woo, Gun-Jo;Hwang, In-Gyun;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1150-1154
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    • 2009
  • Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.

New Records of Xylaria Species in Korea: X. ripicola sp. nov. and X. tentaculata

  • Kim, Chang Sun;Jo, Jong Won;Kwag, Young-Nam;Oh, Soon-Ok;Lee, Sle-gee;Sung, Gi-Ho;Han, Jae-Gu;Oh, Junsang;Shrestha, Bhushan;Kim, Sang-Yong;Shin, Chang-Ho;Han, Sang-Kuk
    • Mycobiology
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    • v.44 no.1
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    • pp.21-28
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    • 2016
  • During a Korean mushroom diversity survey from 2011 to 2014, we found one new Xylaria species (X. ripicola sp. nov.) and one Xylaria species that had not been previously observed in Korea (X. tentaculata). To confirm the phylogenetic placement of the new species, we conducted a phylogenetic investigation based on internal transcribed spacer regions of ribosomal DNA sequences. Additionally, the new species, X. ripicola, was subsequently analyzed for RNA polymerase II subunit sequences. We also evaluated the macroscopic and microscopic features of this species. Herein, X. ripicola is described as a new species that was collected from a natural beach habitat and X. tentaculata is formally reported as newly found in Korea.

Two Unrecoreded Species Belonging to Penicillium Section Exilicaulis in South Korea

  • Park, Myung Soo;Kim, Sung Hyun;Lee, Jun Won;Kim, Ji Seon;Cho, Yoonhee;Lim, Young Woon
    • The Korean Journal of Mycology
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    • v.48 no.3
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    • pp.175-185
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    • 2020
  • Penicillium in section Exilicaulis is characterized by non-vesiculate monoverticillate and biverticillate stipes. Species in sect. Exilicaulis are commonly found in soil and plants in terrestrial environments; however, only a few species have been reported in Korea. To investigate the diversity of Penicillium sect. Exilicaulis, Penicillium species were isolated from terrestrial and marine environments. Based on sequence analyses of β-tubulin, calmodulin, and the second largest subunit of RNA polymerase II loci, 19 strains of Penicillium in sect. Exilicaulis were identified as P. citreonigrum, P. citreosulfuratum, P. corylophilum, P. menonorum, P. rubefaciens, P. velutinum, Penicillium sp. 1, and Penicillium sp. 2. Two of them, P. citreonigrum and P. citreosulfuratum, were confirmed to be new to Korea. Molecular phylogenies and detailed descriptions of the two unrecorded species are provided.