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RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • v.7 no.4
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    • pp.181-186
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    • 2009
  • Myotonic dystrophy type 1 (DM1), which is a dominantly inherited neurodegenerative disorder, results from a CTG trinucleotide repeat expansion in the 3'-untranslated region (3'-UTR) of the myotonic dystrophy protein kinase (DMPK) gene. Retention of mutant DMPK (mDMPK) transcripts in the nuclei of affected cells has been known to be the main cause of pathogenesis of the disease. Thus, reducing the RNA toxicity through elimination of the mutant RNA has been suggested as one therapeutic strategy against DM1. In this study, we suggested RNA replacement with a trans -splicing ribozyme as an alternate genetic therapeutic approach for amelioration of DM1. To this end, we identified the regions of mDMPK 3'-UTR RNA that were accessible to ribozymes by using an RNA mapping strategy based on a trans-splicing ribozyme library. We found that particularly accessible sites were present not only upstream but also downstream of the expanded repeat sequence. Repair or replacement of the mDMPK transcript with the specific ribozyme will be useful for DM1 treatment through reduction of toxic mutant transcripts and simultaneously restore wild-type DMPK or release nucleus-entrapped mDMPK transcripts to the cytoplasm.

Adipocyte-Related Genes and Transcription Factors were affected by Ganoderma lucidum during 3T3-L1 Differentiation (지방세포 분화중인 3T3-L1 세포에서 영지추출물에 의한 지방관련 유전자와 전사인자의 발현 조절)

  • Lee, Chae-Woo;Yoon, Hyun-Min;Kang, Kyung-Hwa
    • Journal of Pharmacopuncture
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    • v.12 no.2
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    • pp.13-20
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    • 2009
  • In this study, the effects of Ganoderma lucidum (GL) on fat metabolism were performed in 3T3-L1 adipocytes. The effects of GL on 3T3-L1 preadipocytes differentiation were also examined. Our results showed that GL decreased the TG content by ORO staining. To elucidate the mechanism of the effects of GL on lowering TG content in 3T3-L1 adipocytes, we examined whether GL modulate the expressions of transcription factors and adipokines related to control of energy expenditure process because adipokines regulate adipocyte mass and increased expenditure may consume much TG in adipocytes. As a result, the expression of C/$EBP{\beta}$, C/$EBP{\delta}$, C/$EBP{\alpha}$, and $PPAR{\gamma}$, genes, which induce the adipose differentiation and adipose-specific FAS, aP2, and adipsin genes, which compose fat formation were decreased. In addition, GL increased the expression of leptin, UCP2, adiponectin in 3T3-L1 adipocytes, resulting in energy homeostasis. In conclusion, GL could regulate transcript factor related to induction of adipose differentiation and control TG content by up-regulation of adipokines related to fat burn.

Differential Expression of Cyclic AMP-Response Element Binding Protein Zhangfei (CREBZF) in the Mouse Testis during Postnatal Development

  • Jang, Hoon
    • Development and Reproduction
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    • v.22 no.1
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    • pp.65-72
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    • 2018
  • Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

Altered Sulfate Metabolism of Arabidopsis Caused by Beet Severe Curly Top Virus Infection

  • Lee, Hong-Gun;Park, Sung-Hee;Kim, Dong-Giun;Lee, Taek-Kyun;Yum, Seung-Shic;Auh, Chung-Kyoon;Lee, Suk-Chan
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.355-360
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    • 2005
  • Sulfur, an important component of plants, is regulated by a variety of stresses in sulfate assimilation and metabolism. Increase has been observed in the expression of O-acetylserine(thiol)lyase (OASTL) through two-dimensional electrophoresis with the shoot tips of Arabidopsis infected by beet severe curly top geminivirus (BSCTV). With the three- to six-fold increases in the transcript expression of OASTL, serine acetyltransferase (SAT) and $\gamma$-glutmylcysteine synthetase (GSH) were induced over the mock-inoculated organization in each organization through real-time RT-PCR analysis. The expression of those genes might affect the accumulation of anthocyanin in symptomatic tissues and the induction of abnormal callus-like structures formed by additional cell divisions as typical disease symptoms of BSCTV-infected Arabidopsis. This is the first report to describe the collaborative induction of OASTL, SAT, and GSH in virus-infected plants. The changed expressions of OASTL, SAT, and GSH in Arabidopsis infected with BSCTV raises new aspects regarding the biological function of symptomatic tissues related to sulfate metabolism.

Biosynthesis of Tetrahydrobiopterin in an Aquatic Fungus, Allomyces macrogynus (물곰팡이 Allomyces macrogynus에서 Tetrahydrobiopterin의 생합성)

  • Lee, Soo Woong;Park, Young Shik
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.243-247
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    • 1998
  • Comparative analyses of intracellular biopterin contents and its biosynthetic enzymes were performed in Allomyces macrogynus. Biopterin content in fresh weight was 14-fold higher in mycelium than in zoospore. Enzyme activities of GTP cyclohydrolase I and 6-pyruvoyltetrahydropterin synthase in ammonium sulfate fractions were approximately 2-fold higher in mycelium. On the other hand, sepiapterin reductase (SR) activity was 10 fold higher in zoospore. Northern blot assay also demonstrated that SR transcript was abundant in zoospore. These results suggest a possible involvement of tetrahydrobiopterin in cellular differentiation of Allomyces macrogyllus as well as provide an experimental basis to elucidate the physiological function of SR in this organism.

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Effects of Kamiyukgunja-tang on anti-CD40 and Recombinant Interleukin-4 induced Cytokine Production and Immunoglobulin E in Highly Purified Mouse B Cells (생쥐의 B 세포에서 면역글로블린 E의 분비와 사이토카인 생산에 대한 가미육군자탕의 효과)

  • Kim Woon Gil;Kim Dong Hee;Park Yang Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.1065-1074
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    • 2003
  • In order to evaluate the antiallergic effects of Kamiyukgunja-tang (KYGJT), studies were done. We measured the cytotoxic activity for lung fibroblast cell, cytokines transcript expression, production of INF-γ, IL-10, IL-4, GM-CSF, IL-1 β, TNF-α. IL-5 proliferation of B cell in anti-CD40mAb plus r1L-4 stimulated murine splenic B cells. The results were obtained as follows : 1. KYGJT was not showed cytotoxicity in the fibroblast lung cell. 2. KYGJT increased the gene synthesis of INF-γ, IL-10, GM-CSF(m-RNA). 3. KYGJT decreased the gene synthesis of IL-1β, IL-4, TNF-α, IL-5(m-RNA). 4. KYGJT decreased the appearance of TNF-α significantly. 5. KYGJT decreased the appearance of IgE significantly. 6. KYGJT decreased the proliferation of B cell significantly. 7. KYGJT decreased the appearance of Histamin Release Production significantly. The facts above prove that KYGJT is effective against the allergy. Thus. I think that we should study on this continuously

Molecular characterization of BrRZFPs genes encoding C3HC4 type RING zinc finger protein under abiotic stress from Chinese cabbage (Brassica rapa L.)

  • Jung, Yu Jin;Lee, Kye Dong;Cho, Yong Gu;Nou, Ill Sup;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.102-110
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    • 2013
  • The novel BrRZFPs genes encoding C3HC4-type RING zinc finger protein were identified from FOX (full length cDNA over-expressing) library of Brassica rapa. Ten full-length cDNAs obtained from the library encode zinc-finger protein containing 346 amino acids, designated BrRZFPs. These genes were classified into four groups by phylogenic analysis showing conserved protein sequences at both termini. The tissue distribution of BrRZFPs transcription was examined by qRT-PCR revealing ubiquitous expression pattern. However, each gene was strongly expressed in the specific tissue. Transcriptional analysis showed that those acquired 10 genes were inducible under abiotic stresses. Likewise, the transcript of BrRZFP3 was strongly induced (~12-folds) by exogenous abscisic acid, whereas the transcripts of BrRZFP1, BrRZFP2 and BrRZFP3 were (> 9-folds) induced by cold. We suggest that these BrRZFPs that function as signal or response to abiotic stress are useful for crop improvement.

Attenuated Expression of Interferon-induced Protein Kinase PKR in a Simian Cell Devoid of Type I Interferons

  • Park, Se-Hoon;Choi, Jaydo;Kang, Ju-Il;Choi, Sang-Yun;Hwang, Soon-Bong;Kim, Jungsuh P.;Ahn, Byung-Yoon
    • Molecules and Cells
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    • v.21 no.1
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    • pp.21-28
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    • 2006
  • The interferon-induced, double-stranded RNA (dsRNA)-dependent protein kinase PKR plays a key role in interferon-mediated host defense against viral infection, and is implicated in cellular transformation and apoptosis. We have isolated a cDNA of simian PKR encoding a product with 83% amino acid identity to the human homolog and showed that PKR expression is significantly attenuated in the Vero E6 African green monkey kidney cells devoid of type I interferon genes. A variant form of PKR lacking the exon 12 in the kinase domain is produced in these cells, presumably from an alternatively spliced transcript. Unlike wild type PKR, the variant protein named PKR-${\Delta}E12$ is incapable of auto-phosphorylation and phosphorylation of eIF2-${\alpha}$, indicating that the kinase sub-domains III and IV embedded in exon 12 are indispensable for catalytic function. PKR-${\Delta}E12$ had no dominant negative effect but was weakly phosphorylated in trans by wild type PKR.

march5 Governs the Convergence and Extension Movement for Organization of the Telencephalon and Diencephalon in Zebrafish Embryos

  • Jung, Jangham;Choi, Issac;Ro, Hyunju;Huh, Tae-Lin;Choe, Joonho;Rhee, Myungchull
    • Molecules and Cells
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    • v.43 no.1
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    • pp.76-85
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    • 2020
  • MARCH5 is a RING finger E3 ligase involved in mitochondrial integrity, cellular protein homeostasis, and the regulation of mitochondrial fusion and fission. To determine the function of MARCH5 during development, we assessed transcript expression in zebrafish embryos. We found that march5 transcripts were of maternal origin and evenly distributed at the 1-cell stage, except for the mid-blastula transition, with expression predominantly in the developing central nervous system at later stages of embryogenesis. Overexpression of march5 impaired convergent extension movement during gastrulation, resulting in reduced patterning along the dorsoventral axis and alterations in the ventral cell types. Overexpression and knockdown of march5 disrupted the organization of the developing telencephalon and diencephalon. Lastly, we found that the transcription of march5 was tightly regulated by the transcriptional regulators CHOP, C/EBPα, Staf, Znf143a, and Znf76. These results demonstrate the essential role of March5 in the development of zebrafish embryos.

A Basic Study on the Development of a Grading Scale of Discourse Competence in Korean Speaking Assessment -Focusing on the Scale of 'REFUSAL' Task (한국어 말하기 평가에서 '담화 능력' 등급 기술을 위한 기초 연구 -'부탁'에 대한 '거절하기' 과제를 중심으로-)

  • Lee, Haeyong;Lee, Hyang
    • Journal of Korean language education
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    • v.29 no.3
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    • pp.255-292
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    • 2018
  • Most grading scales of Korean language proficiency tests are based on existing grading scales that are not empirically verified. The purpose of this study is to develop an empirically verified scale descriptor. The 'Performance data-driven approach' that is suggested by Fulcher (1987) was used to develop the detailed description of characteristics for each level of performance. This study is focused on the functional phase of speech samples analysis (coding data) to create explanatory categories of discourse skills into which individual observations of speech phenomena can be scored. The speech samples that were collected through this study demonstrated stages of speech that can be a foundation of a grading scale. The data used in the study was collected from 23 native speakers of Korean. Speech samples were recorded from simulated speaking tests using the 'REFUSAL' task, and transcribed for analysis. The transcript was analyzed using discourse analysis. The result showed that the 'REFUSAL' task needs to go through four functional phases in actual communication. Furthermore, this study found specific and detailed explanatory categories of discourse competence based on the actual native speaker's speech data. Such findings are expected to contribute to the development of more valid and reliable speaking assessment.