• Title/Summary/Keyword: transcriptional analysis

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Molecular characterization of a novel rice(Oryza sativa L.) MAP kinase, OsEDRl, its role in defense signaling pathway.

  • Kim, Jung-A;Jwa, Nam-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.82-83
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    • 2003
  • Plants have evolved differently from animals having mobile activities. Thus, plants should have developed unique defense mechanisms against biotic/abiotic stresses to which plants are differently exposed, according to seasons. Most organisms have an conserved signaling network using mitogen-activated protein kinase (MAPK) cascade(s). The phenomenon implied that they are functionally very important in all organisms. In fact, they constitute one of the major components of signaling pathways involved in regulating a wide range of cellular activities from growth and development to cell death. Recently, complete MAPK cascade was first characterized in Arabidopsis from the receptor kinase (FLS2) through fellowing MEKKI -MKK4/MKK5-MPK3/MPK6-WRKY22/MRKY29 pathway. Whereas, MAPK cascade signaling pathway in monocot plant including rice (0ryza sativa L.), the most important of all food crops and an established monocot plant research model, MAPKinase kinase kinases (MAPKKK) of rice are the first upstream component of the MAPK cascade, but MAPKKK has been first identified and characterized in our lab and designated as, OsEDRl based on its homology with the Arabidopsis EDRI. The Arabidopsis EDRl was regarded as a negative regulator of defense response and the role of rice OsEDRl was analyzed. Transcriptional regulation of OsEDRl was detected under various stresses and immunoblotting analysis is going on to detect the level of OsEDRl protein in the mutants showing unique phenotype. We also introduced the constitutively active and the dominant negative forms of the OsEDRl for characterizing biological function.

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Leptin potentiates Prevotella intermedia lipopolysaccharide-induced production of TNF-$\alpha$ in monocyte-derived macrophages

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.40 no.3
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    • pp.119-124
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    • 2010
  • Purpose: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-$\alpha$ production in differentiated THP-1 cells, a human monocytic cell line. Methods: LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-$\alpha$ and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-$\alpha$ and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. Results: Leptin enhanced P. intermedia LPS-induced TNF-$\alpha$ production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-$\alpha$ expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-$\alpha$ production was not mediated by the leptin receptor. Conclusions: The ability of leptin to enhance P. intermedia LPS-induced TNF-$\alpha$ production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

Up-regulation of Aldo-keto Reductase 1C3 Expression in Sulforaphane-treated MCF-7 Breast Cancer Cells

  • Lee, Sang-Han
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1079-1085
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    • 2008
  • The chemopreventive activity of sulforaphane (SFN) occurs through its inhibition of carcinogen-activating enzymes and its induction of detoxification enzymes. However, the exact mechanisms by which SFN exerts its anti-carcinogenic effects are not fully understood. Therefore, the mechanisms underlying the cytoprotective effects of SFN were examined in MCF-7 breast cancer cells. Exposure of cells to SFN (10 ${\mu}M$) induced a transcriptional change in the AKR1C3 gene, which is one of aldo-keto reductases (AKRs) family that is associated with detoxification and antioxidant response. Further analysis revealed that SFN elicited a dose- and time-dependent increase in the expression of both the NRF2 and AKR1C3 proteins. Moreover, this up-regulation of AKR1C3 was inhibited by pretreatment with antioxidant, N-acetyl-L-cysteine (NAC), which suggests that the up-regulation of AKR1C3 expression induced by SFN involves reactive oxygen species (ROS) signaling. Furthermore, pretreatment of cells with LY294002, a pharmacologic inhibitor of phosphatidylinositol 3-kinase (PI3K), suppressed the SFN-augmented Nrf2 activation and AKR1C3 expression; however, inhibition of PKC or MEK1/2 signaling with $G\ddot{o}6976$ or PD98059, respectively, did not alter SFN-induced AKR1C3 expression. Collectively, these data suggest that SFN can modulate the expression of the AKR1C3 in MCF-7 cells by activation of PI3K via the generation of ROS.

Transcriptome Analysis of the Barley-Rhynchosporium secalis Interaction

  • Al-Daoude, Antonious;Shoaib, Amina;Al-Shehadah, Eyad;Jawhar, Mohammad;Arabi, Mohammad Imad Eddin
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.425-431
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    • 2014
  • Leaf scald caused by the infection of Rhynchosporium secalis, is a worldwide crop disease resulting in significant loss of barley yield. In this study, a systematic sequencing of expressed sequence tags (ESTs) was chosen to obtain a global picture of the assembly of genes involved in pathogenesis. To identify a large number of plant ESTs, which are induced at different time points, an amplified fragment length polymorphism (AFLP) display of complementary DNA (cDNA) was utilized. Transcriptional changes of 140 ESTs were observed, of which 19 have no previously described function. Functional annotation of the transcripts revealed a variety of infection-induced host genes encoding classical pathogenesis-related (PR) or genes that play a role in the signal transduction pathway. The expression analyses by a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) revealed that Rar1 and Rpg4 are defense inducible genes, and were consistent with the cDNA-AFLP data in their expression patterns. Hence, the here presented transcriptomic approach provides novel global catalogue of genes not currently represented in the EST databases.

Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi

  • Kong, Hyun-Hee;Hwang, Mee-Yeul;Kim, Hyo-Kyung;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.151-160
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    • 2001
  • Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface Proteins , enzymes for DNA, energy Production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba.

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Arg243, Invariably Critical for the Transcriptional Activation of Yeast Gcn4p

  • Cho, Gyu-Chull;Lee, Jae-Yung;Kim, Joon
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.154-158
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    • 1999
  • The arginine residue at position 243 (Arg 243) of the yeast transcription factor, Gcn4p, is invariably conserved among bZIP transcription factors. Using site-directed oligonucleotide saturation mutagenesis involving two-step polymerase chain reaction (PCR) amplification, random mutations were successfully introduced at the codon of 243 in the basic domain of Gcn4p. This mutant library was transformed ito Gcn4p defective yeast strain and selected for the transcriptionally active colonies. All colonies which were transcriptionally active had arginines in the codon 243. In this study, the strand preference by Taq polymerase during mutagenesis was also tested. Oligonucleotides were specially designed to test whether or not the polymerase was preferred using the strand as a template. A population of randomly mutated products were cloned into an appropriate vector and characterized by DNA sequencing analysis. Saturation mutagenesis which was performed efficiently by this method revealed a strong bias in terms of strand preference of Taq polymerase by an approximate ratio of 3 to 1 in this study.

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Molecular Biological Study of The Effects of Gilgyung-Tang(GGT) on Cellular Proliferation and Viability of Normal Human Lung Fibriblast Cell (길경탕(桔梗湯)이 인체(人體) 폐세포(肺細胞)에 미치는 영향(影響)에 관(關)한 분자생물학적(分子生物學的) 연구(硏究))

  • Rhee, Hyung-Koo
    • The Journal of Korean Medicine
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    • v.20 no.2
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    • pp.88-97
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    • 1999
  • To characterize the effects of Gilgyung-Tang(GGT) on cellular proliferation and viability of normal lung fibroblast cells, we examined the cell cycle progression and cell cycle-related gene expression in T3891 using a flow cytometry and a quantitative RT-PCR analysis. 1. The significant surpression effect of cellular proliferations of GGT was observed in proportion to a certain concentration and time. 2. GGT was identified to induce apoptotic death of damaged cells by treatment with a DNA-damage agent and etoposide, while it stimulated the recovery of cellular viability of normal cells. 3 The significant reductions of mRNA expression of PCAN, c-Fos treated by GGT were observed. 4. The significant inductions of mRNA expression of p53, CDKN1. Gadd45 treated by GGT were observed. 5. The apoptosis caused by the reduction of Bcl-2 genes was significant and the Bax genes were increased. but the amount of Fas genes were not changed. These results strongly suggest that GGT triggers arrest of the cell cycle at G1 phase, and thus causes an inhibition of cellular proliferation of human normal lung cells through the transcriptional up-regulation of cell cycle inhibitory genes and down-regulation of induction of cell cycle stimulating genes respectably.

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Photoperiodic Proteins in Plant Cells (식물세포의 일주기성 단백질)

  • Hwang, Hee-Youn;Bhoo, Seong-Hee
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.121-125
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    • 2010
  • In the past 10 years, a lot of plant circadian rhythm researches have published in molecular biology and biochemistry. We discussed with published molecular studies of circadian clock and rhythmic genes in Arabidopsis, rice and algae. However past this studies are not sufficient to explain the whole rhythmic metabolism. Recently many researchers have concerned post-transcriptional, translational and post-translational modification of rhythmic proteins. From the view point of the high-throughput study, we could suggest the proteomic analysis with 2-DE gel electrophoresis and MS/MS techniques for the identification of modified proteins.

Identification and Transcriptional Analysis of Priming Genes in Arabidopsis thaliana Induced by Root Colonization with Pseudomonas chlororaphis O6

  • Cho, Song-Mi;Park, Ju-Yeon;Han, Song-Hee;Anderson, Anne J.;Yang, Kwang-Yeol;Gardener, Brian Mcspadden;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.27 no.3
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    • pp.272-279
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    • 2011
  • Root colonization of Arabidopsis thaliana with Pseudomonas chlororaphis O6 induces systemic tolerance against diverse pathogens, as well as drought and salt stresses. In this study, we demonstrated that 11 genes in the leaves were up-regulated, and 5 genes were down-regulated as the result of three- to five-days root colonization by P. chlororaphis O6. The identified priming genes were involved in cell signaling, transcription, protein synthesis, and degradation. In addition, expression of selected priming genes were induced in P. chlororaphis O6-colonized plants subjected to water withholding. Genes encoding defense proteins in signaling pathways regulated by jasmonic acid and ethylene, such as VSP1 and PDF1.2, were additional genes with enhanced expression in the P. chlororaphis O6-colonized plants. This study indicated that the expression of priming genes, as well as genes involved in jasmonic acid- and ethylene-regulated genes may play an important role in the systemic induction of both abiotic and biotic stress due to root colonization by P. chlororaphis O6.

Endosulfan Induces CYP1A1 Expression Mediated through Aryl Hydrocarbon Receptor Signal Transduction by Protein Kinase C

  • Han, Eun Hee;Kim, Hyung Gyun;Lee, Eun Ji;Jeong, Hye Gwang
    • Toxicological Research
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    • v.31 no.4
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    • pp.339-345
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    • 2015
  • CYP1A1 is a phase I xenobiotic-metabolizing enzyme whose expression is mainly driven by AhR. Endosulfan is an organochlorine pesticide used agriculturally for a wide range of crops. In this study, we investigated the effect of endosulfan on CYP1A1 expression and regulation. Endosulfan significantly increased CYP1A1 enzyme activity as well as mRNA and protein levels. In addition, endosulfan markedly induced XRE transcriptional activity. CH-223191, an AhR antagonist, blocked the endosulfan-induced increase in CYP1A1 mRNA and protein expression. Moreover, endosulfan did not induce CYP1A1 gene expression in AhR-deficient mutant cells. Furthermore, endosulfan enhanced the phosphorylation of calcium calmodulin (CaM)-dependent protein kinase (CaMK) and protein kinase C (PKC). In conclusion, endosulfan-induced up-regulation of CYP1A1 is associated with AhR activation, which may be mediated by PKC-dependent pathways.