• BMB Reports
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• 제43권12호
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• pp.807-812
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• 2010

$NF{\kappa}B$ and ZAS3 are transcription factors that control important cellular processes including immunity, cell survival and apoptosis. Although both proteins bind the ${\kappa}B$-motif, they produce opposite physiological consequences; $NF{\kappa}B$ activates transcription, promotes cell growth and is often found to be constitutively expressed in cancer cells, while ZAS3 generally represses transcription, inhibits cell proliferation and is downregulated in some cancers. Here, we show that ZAS3 inhibits $NF{\kappa}B$-dependent transcription by competing with $NF{\kappa}B$ for the ${\kappa}B$-motif. Transient transfection studies show that N-terminal 645 amino acids is sufficient to repress transcription activated by $NF{\kappa}B$, and that the identical region also possesses intrinsic repression activity to inhibit basal transcription from a promoter. Finally, in vitro DNA-protein interaction analysis shows that ZAS3 is able to displace $NF{\kappa}B$ by competing with $NF{\kappa}B$ for the ${\kappa}B$-motif. It is conceivable that ZAS3 has therapeutic potential for controlling aberrant activation of $NF{\kappa}B$ in various diseases.

• BMB Reports
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• 제30권5호
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• pp.362-369
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• 1997

Multiple phosphorylation of the carboxy-terminal domain (CTD) of the largest subunit in RNA polymerase II (RNAP II) is thought to play an important role in the transcription cycle. The preinitiation complex in a partially purified complete transcription system suggested that RNA polymerase IIA containing unphosphorylated CTD is involved in complex assembly, whereas RNA polymerase IIO containing Ser and Thr phosphorylated CTD is not involved in preinitiation complex assembly. Recently a minimal transcription system was developed which requires chemically defined minimal components for its transcription: TBP, TFIIB, TFIIF, RNAP II and a supercoiled adenovirus-2 major late promoter (Ad-2 MLP). It would be using interesting to determine the consequence of CTD phosphorylation on preinitiation complex formation using the minimal transcription system. Contrary to the results from the partially purified complete transcription system, both RNA polymerase IIA and IIO are equally recruited in the preinitiation complex formation. The discrepancy may result from the two different assays used to determine complex formation, the use of chemically undefined complete and defined minimal transcription systems. This implicates that some factors in the complete transcription system are involved in the distinction between RNAP IIA and IIO in complex assembly. In addition multiple tyrosine phosphorylation of the CTD of RNAP II was prepared with c-Abl kinase and its recruiting ability in the preinitiation complex was examined. Compare with Ser and Thr phosphorylated RNAP IIO, Tyr phosphorylated RNAP IlOy forms a stable preinitiation complex in both the minimal and complete transcription systems. Based on these results, it seems that tyrosine phosphorylation of the CTD is important in the transcription cycle on the special subset of class-II promoter or has a different role in the transcription process.

• 한국자기공명학회논문지
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• 제24권4호
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• pp.117-122
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• 2020

Transcription factors are proteins that bind specific sites or elements in regulatory regions of DNA, known as promoters or enhancers, where they control the transcription or expression of target genes. MEIS1 protein is a DNA-binding domain present in human transcription factors and plays important roles in various biological functions. The hydrogen exchange rate constants of the imino protons were determined for the wild-type containing the consensus DNA-binding site for the MEIS1 and those of the mutant DNA duplexes using NMR spectroscopy. The G2A-, A3G- and C4T-mutant DNA duplexes lead to clear changes in thermal stabilities of these four consensus base pairs. These unique dynamic features of the four base pairs in the consensus 5'-TGAC-3' sequence might play crucial roles in the effective DNA binding of the MEIS1 protein.

• 생약학회지
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• 제44권2호
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• pp.104-109
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• 2013

Hesperidin (HES), a flavonone glycoside isolated from the citrus fruits such as lemons and oranges, has been reported to have many biological properties including antiinflammatory, antioxidant, and antiallergy activities. In this study, we focused on the action of HES modulating Th2-associated cytokines such as IL-4 and IL-13 expression in PMA/ionomycin (PI)-stimulated rat basophilic leukemia (RBL-2H3) cells. The production of IL-4 and IL-13 was quantified by ELISA and the mRNA expression was detected by using RT-PCR assay. In addition, western blot analysis was performed to determine the transcription factors involved in the cytokine expression. We found that HES significantly decreased PI-induced IL-4 and IL-13 productions and also decreased the level of mRNA in a dose-dependent manner. Furthermore, western blot analysis of the transcription factors implied that HES down-regulated the protein level of c-Jun and c-Fos, which are the activating protein 1 (AP-1) family and nuclear factor-kappaB (NF-${\kappa}B$) characterized as a transcription factors related to the Th2-associated cytokine expression. Taken together, our data showed that the action of HES responsible for antiallergy activities is based on suppression of Th2-associated cytokines through inhibition of AP-1 and NF-${\kappa}B$ transcription factors.

• 대한한방소아과학회지
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• 제26권3호
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• pp.30-45
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• 2012

Objectives The suppressive effect of CSBHT has been mysterious. Thus, the present study is designed to investigate the suppressive effect and its mechanism. Methods To investigate the anti-allergy effect from ChungShimBoHyeolTang(CSBHT), RBL-2H3 cell was used and examined by Real-Time PCR, and IL-4 and IL-13 from RBL-2H3 was examined by ELIS. In addition, GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, c-Jun, NF-${\kappa}B$ p65 transcription factors of RBL-2H3 mast cell were examined by Western Blotting. Also, OVA/alum-sensitized mice were orally administrated CSBHT and serum OVA-specific IgE production, IL-4, and IL-13 production in splenocytes supernatant were examined. Results As a result of treating with CSBHT extract, RBL-2H3 mast cells significantly suppressed the IL-4 and IL-13 mRNA expression and IL-4 and IL-13 production. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, and NF-${\kappa}B$ p65. Also, examining the mice, administration of CSBHT suppressed the amount of OVA-specific IgE in OVA/alum-sensitized mice and IL-4 and IL-13 production in splenocytes supernatant. Conclusions The study suggested that the anti-allergic activities of CSBHT suppresses IL-4 and IL-13 production from the Th2 cytokines by suppressing transcription factors as GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 in mast cells.

• 대한한방소아과학회지
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• 제25권1호
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• pp.28-39
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• 2011

Objectives: SaengRyoSaMulTang is a herbal formula in Oriental Medicine, known anti-allergens. However, its mechanism and cellular targets have not been found yet. Thus the study has developed to investigate the suppressive effect of SaengRyoSaMulTang. Methods: In the study, cellular viability, IL-4, IL-13 mRNA expression, IL-4, IL-13 production, manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}$B p65 transcription factors were examined by Real-Time PCR, ELISA analysis and western blotting. Results: As a result of treating with SaengRyoSaMulTang extract(SRSMT), the study has shown that the amount of Th2 cytokines, which include PI induced IL-4 and IL-13, plays a significant role in suppressing effect. RBL-2H3 mast cells significantly suppressed the PI-induced Th2 cytokine production including IL-4 and IL-13 in a dose dependent manner. PI-induced IL-4 and IL-13 production was significantly suppressed by SRSMT intervention. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos, but NF-${\kappa}$B p65. Conclusions: The study suggests that the anti-allergenic activities of SRSMT may regulate the transcription factors GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos inhibiting Th2 cytokines IL-4 and IL-13 in mast cells.

• 대한통합의학회지
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• 제11권3호
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• pp.59-67
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• 2023

Purpose : Epithelial-to-mesenchymal transition (EMT) is recognized as an important cellular response in metastatic proceduresand characterized by loss of cellular polarity as well as gain of mesenchymal features, which enables migration and invasion. Hepatocellular carcinoma (HCC) is one of the most common primary carcinomas in the liver and exhibits a poor prognosis due to frequent extrahepatic metastasis. Taraxacum officinale has been used for a long time in oriental medicine because of its various pharmacological activitiessuch as anti-rheumatic, anti-inflammatory, antioxidative, and anticarcinogenic activities. In this study, the anti-metastatic activity of T. officinale water extract (TOWE) and ethanol extract (TOEE) was investigated through the regulation of EMT in the Huh7 cells. Methods : The effects of TOWE and TOEE on migratory and invasive activities were investigated by wound healing and in vitro invasion assays. Western blot analysis was also applied to analyze protein expression levels associated with EMT and their upstream transcription factors in Huh7 cells. Results : TOWE and TOEE treatment potently inhibited migration and invasion of Huh7 cells compared to the untreated group. Both extracts treatment inhibited protein expression levels of N-cadherin, matrix metalloproteinase (MMP)-9, and vimentin while E-cadherin was significantly accelerated. In addition, the activated status of transcription factors, Snail, nuclear factor (NF)-κ B, and zinc finger E-box binding homeobox (ZEB)1 was also inhibited with statistical significance. In comparison to both extracts, TOEE more potently attenuated migration, invasion, and EMT markers as well as their transcription factors in Huh7 cells than TOWE, which means that TOEE might possess more functional phytochemicals than TOWE. Conclusion : Consequently, TOWE and TOEEattenuated metastatic activity of hepatocellular carcinoma through the regulation of EMT markers and their transcription factors in Huh7 cells, which means that T. officinale might be a promising strategy for a chemopreventive agent against HCC metastasis.

• 한국초지조사료학회지
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• 제41권4호
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• pp.242-249
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• 2021

Forage crop management is severely challenged by global warming-induced climate changes representing diverse a/biotic stresses. Thus, screening of valuable genetic resources would be applied to develop stress-tolerant forage crops. We isolated two NAC (NAM, ATAF1, ATAF2, CUC2) transcription factors (ANAC032 and ANAC083) transcriptionally activated by multi-abiotic stresses (salt, drought, and cold stresses) from Arabidopsis by microarray analysis. The NAC family is one of the most prominent transcription factor families in plants and functions in various biological processes. The enhanced expressions of two ANACs by multi-abiotic stresses were validated by quantitative RT-PCR analysis. We also confirmed that both ANACs were localized in the nucleus, suggesting that ANAC032 and ANAC083 act as transcription factors to regulate the expression of downstream target genes. Promoter activities of ANAC032 and ANAC083 through histochemical GUS staining again suggested that various abiotic stresses strongly drive both ANACs expressions. Our data suggest that ANAC032 and ANAC083 would be valuable genetic candidates for breeding multi-abiotic stress-tolerant forage crops via the genetic modification of a single gene.

• Molecules and Cells
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• 제40권3호
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• pp.169-177
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• 2017

Tissue-specific transcription is critical for normal development, and abnormalities causing undesirable gene expression may lead to diseases such as cancer. Such highly organized transcription is controlled by enhancers with specific DNA sequences recognized by transcription factors. Enhancers are associated with chromatin modifications that are distinct epigenetic features in a tissue-specific manner. Recently, super-enhancers comprising enhancer clusters co-occupied by lineage-specific factors have been identified in diverse cell types such as adipocytes, hair follicle stem cells, and mammary epithelial cells. In addition, noncoding RNAs, named eRNAs, are synthesized at super-enhancer regions before their target genes are transcribed. Many functional studies revealed that super-enhancers and eRNAs are essential for the regulation of tissue-specific gene expression. In this review, we summarize recent findings concerning enhancer function in tissue-specific gene regulation and cancer development.

• Journal of Plant Biotechnology
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• 제49권4호
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• pp.292-2999
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• 2022

Anthocyanin, an important component in the grape berry skin, strongly affects grape quality. The transcription factors VvMYBA1 and VvMYBA2 (VvMYBA1/2) control anthocyanin biosynthesis. In addition, cultivation and environmental factors, such as light, influence anthocyanin accumulation. The present study aimed to clarify the effect of shading (reduced light condition) on the transcriptomic regulation of anthocyanin biosynthesis using a red-wine grape cultivar, Vitis vinifera 'Pinot Noir', and its white mutant, 'Pinot Blanc', caused by the deletion of the red allele of VvMYBA1/2. The grape berry skins were analyzed for anthocyanin content and global gene transcription accumulation. The microarray data were later validated by quantitative real-time PCR. A decisive influence of VvMYBA1/2 on the expression of an anthocyanin-specific gene, UDP glucose: flavonoid 3-O-glucosyltransferase, was observed as expected. In contrast, upstream genes of the pathway, which are shared by other flavonoids, were also expressed in 'Pinot Blanc', and the mRNA levels of some of these genes decreased in both cultivars on shading. Thus, the involvement of light-sensitive transcription factor(s) other than VvMYBA1/2 was suggested for the expression control of the upstream genes of the anthocyanin biosynthetic pathway. Furthermore, it was suggested that the effects of these factors are different among isogenes.