• 제목/요약/키워드: the transcription factors

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Analysis of Bovine Interferon-tau Gene subtypes Expression in the Trophoblast and Non-trophoblast cells

  • Kim, Min-Su;Lim, Hyun-Joo;Lee, Ji Hwan;Park, Soo Bong;Won, Jeong-Il;Kim, Hyun Jong
    • 한국수정란이식학회지
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    • 제33권4호
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    • pp.195-203
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    • 2018
  • Interferon-tau (IFNT) is known as a major conceptus protein that signals the process of maternal recognition of pregnancy in ruminants. Also, multiple interferon genes exist in cattle, However, molecular mechanisms of these bovine IFNT (bIFNT) genes whose expressions are limited have not been characterized. We and others have observed that expression levels of bovine subtype IFNT genes in the tissues of ruminants; thus, bIFNT1 and other new type I (bIFNTc1/c2/c3) gene co-exist during the early stages of conceptus development and non-trophoblast cells. Its genes transcription could be regulated through CDX2 and ETS2 and JUN and/or cAMP-response element binding protein (CREB)-binding protein (CREBBP) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. Bovine ear-derived fibroblast cells, were co-transfected with luciferase reporter constructs carrying upstream (positions -1000 to +51) regions of bIFNT1 and other new type I gene and various transcription factor expression plasmids. Compared to each - 1kb-bIFNT1/c1/c2/c3-Luc increased when this constructs were co-transfected with CDX2, ETS2, JUN and/or CREBBP. Also, Its genes was had very effect on activity by CDX2, either alone or with the other transcription factors, markedly increased luciferase activity. However, the degree of transcriptional activation of the bIFNTc1 gene was not similar to that bIFNT1/c2/c3 gene by expression plasmid. Furthermore, Sequence analyses also revealed that the expression levels of bIFNT1/c2/c3 gene mRNAs expression were highest on day 17, 20 and 22 trophoblast and, Madin-Darby bovine kidney (MDBK), Bovine ear-derived fibroblast (EF), and endometrium (Endo) non-trophoblast cells. But, bIFNTc1 mRNA had not same expression level, bIFNTc1 lowest levels than those of IFNT1/c2/c3 gene in both trophoblast and non-trophoblast cells. These results demonstrate that bovine subtype bIFNT genes display differential, in the trophoblast and non-trophoblast cells.

금앵자 에탄올 추출물에 의한 3T3-L1 지방세포의 분화억제 효과와 그 메커니즘 규명 (Inhibitory Effects and Molecular Mechanism of Adipocyte Differentiation by Rosae laevigata Fructus Ethanol Extracs)

  • 정현영;정인교;남소연;윤희정;김병우;권현주
    • 한국미생물·생명공학회지
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    • 제44권1호
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    • pp.89-97
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    • 2016
  • 비만은 체내 지방이 과도하게 축적되어 일어나는 현상으로, 당뇨, 고혈압, 심혈관 질환 및 암과 같은 질병의 원인이 된다. 본 연구는 RLE에 의해 지방전구세포에서 지방세포로 분화 시, 세포 내 축적되는 Triglyceride 저해 및 발현되는 전사인자들의 발현양상에 미치는 영향에 대하여 조사하였다. 그 결과, RLE는 Oil Red O 염색에서 세포 내 triglyceride의 축적을 농도 의존적으로 억제하였다. 또한 CCAAT/enhancer binding protein(C/EBP) ${\alpha}$, ${\beta}$와 peroxisome proliferator activated receptor ${\gamma}$($PPAR{\gamma}$)과 같은 지방세포 분화 관련 전사인자들의 발현을 억제하였다. RLE는 clonal expansion 단계의 지방세포를 G1기에서 세포 주기를 정지시켜 세포의 증식을 억제하였으며, RLE 처리에 의해 p21의 증가, Cyclin E, Cdk2, Phospho-Rb의 발현 저해 등 G1 arrest 관련 단백질의 발현 변화가 유도되었다. 따라서, RLE는 분화 관련 전사인자들의 발현을 조절하고 지방세포 분화 초기에 G1기의 세포 주기 정지를 억제함으로써 지방전구세포에서 지방세포로의 분화를 억제한다고 사료된다.

The Up-Regulation of miR-199b-5p in Erythroid Differentiation Is Associated with GATA-1 and NF-E2

  • Li, Yuxia;Bai, Hua;Zhang, Zhongzu;li, Weihua;Dong, Lei;Wei, Xueju;Ma, Yanni;Zhang, Junwu;Yu, Jia;Sun, Guotao;Wang, Fang
    • Molecules and Cells
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    • 제37권3호
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    • pp.213-219
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    • 2014
  • MicroRNAs (miRNAs) represent a class of small non-coding regulatory RNAs that play important roles in normal hematopoiesis, including erythropoiesis. Although studies have identified several miRNAs that regulate erythroid commitment and differentiation, we do not understand the mechanism by which the crucial erythroid transcription factors, GATA-1and NF-E2 directly regulate and control differentiation via miRNA pathways. In this study, we identified miR-199b-5p as a key regulator of human erythropoiesis, and its expression was up-regulated during the erythroid differentiation of K562 cells. Furthermore, the increase of miR-199b-5p in erythroid cells occurred in a GATA-1- and NF-E2-dependent manner during erythrocyte maturation. Both GATA-1 and NF-E2 bound upstream of the miR-199b gene locus and activated its transcription. Forced expression of miRNA-199b-5p in K562 cells affected erythroid cell proliferation and maturation. Moreover, we identified c-Kit as a direct target of miR-199b-5p in erythroid cells. Taken together, our results establish a functional link among the erythroid transcription factors GATA-1/NF-E2, miR-199b-5p and c-Kit, and provide new insights into the coupling of transcription and post-transcription regulation in erythroid differentiation.

조릿대 에틸아세테이트 분획물의 지방세포에서 분화전사인자 조절을 통한 지방형성 저해 효능 (Inhibitory Effects of Sasa borealis on Mechanisms of Adipogenesis)

  • 박희숙;김건희
    • 한국식품영양과학회지
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    • 제42권6호
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    • pp.837-843
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    • 2013
  • 본 연구에서는 3T3-L1 지방전구세포를 이용하여 조릿대조추출물(SBE)과 에틸아세테이트 분획물(SBEA)의 지방세포 내 중성지방 축적 저해 활성을 확인하고자 하였다. 먼저 SBE의한 지방세포 분화 저해 활성을 확인하기 위해 추출물을 3T3-L1 지방전구세포에 분화를 유도하면서 농도별(10, 50, 100 ${\mu}g/mL$)로 처리하였고, 그 결과 SBE가 지방세포의 분화를 억제시켜 지방세포 내 중성지방 축적을 저해시켰다. 또한 SBE를 용매 극성에 따른 분획한 분획물들의 항분화 효능을 확인하였다. 그중 항분화 효능이 가장 뛰어난 에틸아세테이트 분획물로 지방세포 분화에 따른 세포 내 중성지방축적이 억제 되었다. 그러나, 지방세포 분해를 통한 glycerol release의 증가는 나타나지 않았다. 이 같은 결과를 바탕으로 항분화 효능의 기전을 연구하기 위해 PPAR${\gamma}$, C/EBP${\alpha}$ 등 전사활성과 지방세포 분화에 관여하는 유전자들의 활성을 확인해 보았다. 실험 결과 SBEA는 PPAR${\gamma}$와 C/EBP${\alpha}$의 mRNA 발현을 농도 의존적으로 감소시켰다. 따라서 SBEA 항비만 효과는 지방 생성의 주요 전사인자인 PPAR${\gamma}$와 C/EBP${\alpha}$의 유전자 발현조절을 통해 지방 분화 억제 및 지방 축적을 효과적으로 감소시키는 것으로 보이며, 효과가 있는 농도가 100 ${\mu}g/mL$로 천연물질로써 비교적 낮은 농도에서 우수한 지방 분화억제 활성을 나타내어 경제적이며 효과적인 항비만 기능성식품으로서 개발 가능성이 있을 것으로 사료된다.

간경화 동물모델에서 Chimeric decoy oligodeoxynucleotide로 억제되는 NF-κB와 Sp1 전사인자 발현 억제 효과에 대한 연구 (Inhibitory Effects of Chimeric Decoy Oligodeoxynucleotide in the Regulation of Transcription Factors NF-κB and Sp1 in an Animal Model of Liver Cirrhosis)

  • 김경현;박지현;김수정;이우람;장영채;김현철;박관규
    • 생명과학회지
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    • 제19권10호
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    • pp.1360-1367
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    • 2009
  • 간섬유화는 지속적인 간세포 손상에 대한 수복현상으로 일어나며, 급성 염증반응과 같은 손상이 주어진 후에는 간세포의 괴사 및 세포외기질의 축적이 일어나게 된다. 간섬유화에 대한 새로운 치료방법을 모색하기 위하여 본 연구에서는 간섬유화 과정에서 염증 반응과 관련된 NF-$\kappa$B와 세포외기질의 축적과 관련된 Sp1전사인자를 동시에 조절하여 간섬유화 억제효과를 관찰하고자 하였다. 전사인자인 Sp1과 NF-$\kappa$B를 동시에 억제하기 위하여 한 분자 내에 Sp1과 NF-$\kappa$B의 전사인자와 결합하는 부위를 가지는 Chimeric (Chi) decoy oligodeoxynucleotide (ODN)을 제작하였다. Chi decoy ODN은 활성화된 간성상세포에서 간섬유화 와 관련된 유전자 발현을 억제시켰으며, 섬유화 동물모델에서도 간 조직의 염증 반응 및 섬유화 관련 인자의 발현을 현저히 억제시켰다. 따라서 Chi decoy ODN은 간섬유화 및 활성화된 간성상세포의 활성을 억제할 수 있는 유전자 치료제로 고려될 수 있을 것으로 사료된다.

Identification of the 187 bp EphA7 Genomic DNA as the Dorsal Midline-Specific Enhancer of the Diencephalon and Mesencephalon

  • Kim, Yujin;Park, Eunjeong;Park, Soochul
    • Molecules and Cells
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    • 제38권11호
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    • pp.1007-1012
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    • 2015
  • EphA7 is a key molecule in regulating the development of the dien- and mesencephalon. To get insight into the mechanism of how EphA7 gene expression is regulated during the dorsal specification of the dien- and mesencephalon, we investigated the cis-acting regulatory sequence driving EphA7 to the dorsal midline of the dien- and mesencephalon. Transgenic LacZ reporter analysis, using overlapping EphA7 BACs, was used to narrow down the dorsal midline-specific enhancer, revealing the 25.3 kb genomic region as the enhancer candidate. Strikingly, this genomic DNA was located far downstream of the EphA7 transcription start site, +302.6 kb to +327.9 kb. Further enhancer mapping, using comparative genomic analysis and transgenic methods, showed that the 187 bp genomic DNA alone, approximately 305 kb downstream of the EphA7 transcription start site, was sufficient to act as the dorsal midline-specific enhancer of EphA7. Importantly, our results indicate that the 187 bp dorsal midline-specific enhancer is critically regulated by homeobox transcription factors during the development of the dien- and mesencephalon.

Glucose and Insulin Stimulate Lipogenesis in Porcine Adipocytes: Dissimilar and Identical Regulation Pathway for Key Transcription Factors

  • Zhang, Guo Hua;Lu, Jian Xiong;Chen, Yan;Dai, Hong Wei;ZhaXi, YingPai;Zhao, Yong Qing;Qiao, Zi Lin;Feng, Ruo Fei;Wang, Ya Ling;Ma, Zhong Ren
    • Molecules and Cells
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    • 제39권11호
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    • pp.797-806
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    • 2016
  • Lipogenesis is under the concerted action of ChREBP, SREBP-1c and other transcription factors in response to glucose and insulin. The isolated porcine preadipocytes were differentiated into mature adipocytes to investigate the roles and interrelation of these transcription factors in the context of glucose- and insulin-induced lipogenesis in pigs. In ChREBP-silenced adipocytes, glucose-induced lipogenesis decreased by ~70%, however insulin-induced lipogenesis was unaffected. Moreover, insulin had no effect on ChREBP expression of unperturbed adipocytes irrespective of glucose concentration, suggesting ChREBP mediate glucose-induced lipogenesis. Insulin stimulated SREBP-1c expression and when SREBP-1c activation was blocked, and the insulin-induced lipogenesis decreased by ~55%, suggesting SREBP-1c is a key transcription factor mediating insulin-induced lipogenesis. $LXR{\alpha}$ activation promoted lipogenesis and lipogenic genes expression. In ChREBP-silenced or SREBP-1c activation blocked adipocytes, $LXR{\alpha}$ activation facilitated lipogenesis and SREBP-1c expression, but had no effect on ChREBP expression. Therefore, $LXR{\alpha}$ might mediate lipogenesis via SREBP-1c rather than ChREBP. When ChREBP expression was silenced and SREBP-1c activation blocked simultaneously, glucose and insulin were still able to stimulated lipogenesis and lipogenic genes expression, and $LXR{\alpha}$ activation enhanced these effects, suggesting $LXR{\alpha}$ mediated directly glucose- and insulin-induced lipogenesis. In summary, glucose and insulin stimulated lipogenesis through both dissimilar and identical regulation pathway in porcine adipocytes.

Epidemiologic Trends of Diarrhea-causing Virus Infection Analyzed by Multiplex Reverse Transcription PCR in Cheonan, Korea, 2010-2018

  • Park, Ji On;Jeon, Jae-Sik;Kim, Jae Kyung
    • 한국미생물·생명공학회지
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    • 제47권2호
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    • pp.317-322
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    • 2019
  • Gastroenteritis with diarrhea is one of the most infectious diseases in the world following respiratory infections. Notably, diarrhea-causing viruses (DVs) cause more than 70% of such cases. In this study, 3,065 stool specimens from patients with diarrhea (median age, 1.1 years; range, 0.0-91.1 years), who were admitted to the DanKook University Hospital, were examined using multiplex reverse transcription PCR (mRT-PCR). The target viruses were astrovirus (AstV), enteric adenovirus (EAdV), group A rotavirus (RotV), norovirus GI (NoV-GI), and norovirus GII (NoV-GII). The mRT-PCR results were analyzed based on various factors such as seasonality, age, presence of co-infection, and analyzed trends. The detection rate of the DVs during the study period was found to be 30.8% (n = 943/3,065). When the detection rate was analyzed monthly, the DV detection rate was found to be highest between December to January. Of the detected DVs, NoV-GII was the most common, accounting for 45.5% of the detected viruses (n = 446/980). Notably, 86.5% (n = 848/980) of the pathogens were detected in individuals who were less than 5 years of age. During the study period, NoV-GII and RotV showed alternating trends. In addition, both the number and rate of co-infections increased.

닭 인터페론 유전자의 클로닝에 관한 연구 (MOLECULAR CLONING OF CHICKEN INTERFERON-GAMMA)

  • 송기덕;;한재용
    • 한국가금학회:학술대회논문집
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    • 한국가금학회 1999년도 제16차 정기총회및학술발표회
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    • pp.34-50
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    • 1999
  • A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

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