• 제목/요약/키워드: regulatory regions

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Genetic Architecture of Transcription and Chromatin Regulation

  • Kim, Kwoneel;Bang, Hyoeun;Lee, Kibaick;Choi, Jung Kyoon
    • Genomics & Informatics
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    • 제13권2호
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    • pp.40-44
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    • 2015
  • DNA microarray and next-generation sequencing provide data that can be used for the genetic analysis of multiple quantitative traits such as gene expression levels, transcription factor binding profiles, and epigenetic signatures. In particular, chromatin opening is tightly coupled with gene transcription. To understand how these two processes are genetically regulated and associated with each other, we examined the changes of chromatin accessibility and gene expression in response to genetic variation by means of quantitative trait loci mapping. Regulatory patterns commonly observed in yeast and human across different technical platforms and experimental designs suggest a higher genetic complexity of transcription regulation in contrast to a more robust genetic architecture of chromatin regulation.

폐특이 전사조절 유전자의 DNAse 1 Hypersensitive Sites (DNAse 1 Hypersensitive Sites of Lung Specific Transcription Factor Gene)

  • 이용철
    • Tuberculosis and Respiratory Diseases
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    • 제48권6호
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    • pp.879-886
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    • 2000
  • 연구배경: 폐특이 전사조절 유전자인 Thyroid Transcription Factor-1 (TTF-1)유전자는 폐에 선택적인 유전자의 표현의 조절에 중요한 전사인자로 작용하고 폐의 발생에서 morphogenic protein으로서 작용한다. 그러나 현재까지 이 TTF-1 유전자의 전사인자에 대한 연구는 거의 미미하다. DNase 1 hypersensitive(DH) regions은 활동적인 염색체에 대한 중요한 표식자이며 유전자를 조절하는 많은 DNA sequences와 밀접한 관계가 있다. 방법 : 추정적인 distal regulatory elements를 밝혀 내기 위해서 TTF-1을 표현하는 인간의 폐선암 세포주인 NCI-H441을 사용해 DNase 1 hypersensitive site assay를 이용하였다. 결과 : TTF-1 유전자에는 전사의 시작부위에서 +150, -450, -800, 그리고 -1500 base pair부위에 4곳의 DH sites가 있음을 할 수 있었다. 결론 : 이상의 결과로 전사 조절부위가 TTF-1 유전자 내에 그리고 5' prime부위에 위치함을 추정할 수 있었다.

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Identification of Putative Regulatory Alterations Leading to Changes in Gene Expression in Chronic Obstructive Pulmonary Disease

  • Kim, Dong-Yeop;Kim, Woo Jin;Kim, Jung-Hyun;Hong, Seok-Ho;Choi, Sun Shim
    • Molecules and Cells
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    • 제42권4호
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    • pp.333-344
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    • 2019
  • Various genetic and environmental factors are known to be associated with chronic obstructive pulmonary disease (COPD). We identified COPD-related differentially expressed genes (DEGs) using 189 samples accompanying either adenocarcinoma (AC) or squamous cell carcinoma (SC), comprising 91 normal and 98 COPD samples. DEGs were obtained from the intersection of two DEG sets separately identified for AC and SC to exclude the influence of different cancer backgrounds co-occurring with COPD. We also measured patient samples named group 'I', which were unable to be determined as normal or COPD based on alterations in gene expression. The Gene Ontology (GO) analysis revealed significant alterations in the expression of genes categorized with the 'cell adhesion', 'inflammatory response', and 'mitochondrial functions', i.e., well-known functions related to COPD, in samples from patients with COPD. Multi-omics data were subsequently integrated to decipher the upstream regulatory changes linked to the gene expression alterations in COPD. COPD-associated expression quantitative trait loci (eQTLs) were located at the upstream regulatory regions of 96 DEGs. Additionally, 45 previously identified COPD-related miRNAs were predicted to target 66 of the DEGs. The eQTLs and miRNAs might affect the expression of 'respiratory electron transport chain' genes and 'cell proliferation' genes, respectively, while both eQTLs and miRNAs might affect the expression of 'apoptosis' genes. We think that our present study will contribute to our understanding of the molecular etiology of COPD accompanying lung cancer.

NEUROD1 Intrinsically Initiates Differentiation of Induced Pluripotent Stem Cells into Neural Progenitor Cells

  • Choi, Won-Young;Hwang, Ji-Hyun;Cho, Ann-Na;Lee, Andrew J.;Jung, Inkyung;Cho, Seung-Woo;Kim, Lark Kyun;Kim, Young-Joon
    • Molecules and Cells
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    • 제43권12호
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    • pp.1011-1022
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    • 2020
  • Cell type specification is a delicate biological event in which every step is under tight regulation. From a molecular point of view, cell fate commitment begins with chromatin alteration, which kickstarts lineage-determining factors to initiate a series of genes required for cell specification. Several important neuronal differentiation factors have been identified from ectopic over-expression studies. However, there is scarce information on which DNA regions are modified during induced pluripotent stem cell (iPSC) to neuronal progenitor cell (NPC) differentiation, the cis regulatory factors that attach to these accessible regions, or the genes that are initially expressed. In this study, we identified the DNA accessible regions of iPSCs and NPCs via the Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq). We identified which chromatin regions were modified after neuronal differentiation and found that the enhancer regions had more active histone modification changes than the promoters. Through motif enrichment analysis, we found that NEUROD1 controls iPSC differentiation to NPC by binding to the accessible regions of enhancers in cooperation with other factors such as the Hox proteins. Finally, by using Hi-C data, we categorized the genes that directly interacted with the enhancers under the control of NEUROD1 during iPSC to NPC differentiation.

CFD APPLICATION TO THE REGULATORY ASSESSMENT OF FAC-CAUSED CANDU FEEDER PIPE WALL THINNING ISSUE

  • Kang, Dong-Gu;Jo, Jong-Chull
    • Nuclear Engineering and Technology
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    • 제40권1호
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    • pp.37-48
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    • 2008
  • Flow fields inside feeder pipes have been simulated numerically using a CFD (computational fluid dynamics) code to calculate the shear stress distribution, which is the most important factor in predicting the local regions of feeder pipes highly susceptible to FAC (flow-accelerated corrosion)-induced wall thinning. The CFD approach, with schemes used in this study, to simulate the flow situations inside the CANDU feeder pipes has been verified as it showed a good agreement between the investigation results for the failed feedwater pipe at Surry unit 2 plant in the U.S. and the CFD calculation. Sensitivity studies of the three geometrical parameters, such as angle of the first and second bends, length of the first span between the grayloc hub and the first bend, and length of the second span between the first and the second bends have been performed. CFD analysis reveals that the local regions of feeder pipes of Wolsung unit 1 in Korea, on which wall thickness measurements have been performed so far, are not coincident with the worst regions predicted by the present CFD analysis located in the connection region of straight and bend pipe near the inlet part of the bend intrados. Finally, based on the results of the present CFD analysis, a guide to the selection of the weakest local positions where the measurement of wall thickness should be performed with higher priority has been provided.

Effective Exon-Intron Structure Verification of a 1-Pyrroline-5-Carboxylate-Synthetase Gene from Halophytic Leymus chinensis (Trin.) Based on PCR, DNA Sequencing, and Alignment

  • Sun, Yan-Lin;Hong, Soon-Kwan
    • 한국자원식물학회지
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    • 제23권6호
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    • pp.526-534
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    • 2010
  • Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.

Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle

  • Baik, M.;Vu, T.T.T.;Piao, M.Y.;Kang, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권10호
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    • pp.1493-1498
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    • 2014
  • Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

Function identification of bovine Nramp1 promoter and intron 1

  • Hao, Linlin;Zhang, Libo;Li, Mingtang;Nan, Wang;Liu, Songcai;Zhong, Jifeng
    • Animal cells and systems
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    • 제15권3호
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    • pp.227-233
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    • 2011
  • The Nramp1/Slc11a1 locus encodes a proton-coupled divalent cation transporter, expressed in late endosomes/lysosomes of macrophages, that constitutes a component of the innate immune response to combat intracellular pathogens and it was shown to play an important role in regulating inherent immunity. The previously identified Z-DNA forming polymorphic repeat(GT)n in the promoter region of the human Nramp1 gene does act as a functional polymorphism influencing gene expression. Research has shown that INF-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$ and bacteria LPS increase the level of Nramp1 expression. However, the molecular mechanism for Nramp1 gene regulation is unclear. In this research, bovine Nramp1 5'-flanking region (-1748~+769) was cloned and analyzed by bioinformatics. Then to find the core promoter and the cis-acting elements, deletion analysis of promoter was performed using a set of luciferase reporter gene constructs containing successive deletions of the bovine Nramp1 5'-flanking regions. Promoter activity analysis by the dual luciferase reporter assay system showed that the core promoter of Nramp1 was located at +58~-89 bp. Some positive regulatory elements are located at -89~-205 bp and -278~-1495 bp. And the repressor elements were in region -205~-278 bp, intron1 and -1495~-1748 bp. LPS-responsive regions were located at -1495~-1748 bp and -278~-205 bp. The present study provides an initial effort to explore the molecular mechanism of transcriptional activation of the bovine Nramp1 gene and should facilitate further studies to decode the complex regulatory process and for molecular breeding for disease resistance in bovines.

Construction and Characterization of Novel Expression Vectors for Genetic Adipose Tissue Ablation

  • Ko, Duck Sung;Choi, Woong Hwan;Kim, Chul Geun
    • Animal cells and systems
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    • 제2권2호
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    • pp.249-258
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    • 1998
  • Obesity, one of the most common metabolic diseases in industrial countries is characterized by an increase in the number or size of adipocytes. In an effort to create transgenic mouse models for the study of obesity we developed a novel technique in which adipose tissue can be ablated genetically at will, at any specific developmental stage and/or physiological condition, by the treatment of ganciclovir. We made a series of adipocytespecific expression vectors using minimal regulatory regions of brown adipocyte-specific uncoupling protein (UCP-1) gene and adipocyte-specific aP2 gene, and then analyzed their expression characteristics in cultured cell lines. When both constructs pUCP-LacZ and paP2-LacZ were transfected transiently into differentiating 3T3-L1 (pre-while adipocytes) and HIB-1B (pre-brown adipocytes) cell lines in vitro and then monitored by X-gal staining of cells, these regulatory regions were sufficient to show proper differentiation stage-specific expression in adipocvtes. To confirm that adipocytes expressing HSV-TK controlled by these minimal requlatory elements are sufficient to kill themselves with ganciclovir treatment pUCP-TK and paP2-TK expression constructs were transfected stably into HIB-1B and 3T3-L1 cells, respectively, and their ganciclovir sensitivities were tested during in vitro differentiation of cells. As expected more than 80% of cells were dead by the 7th day of treatment with ganciclovir while negative control cells were not affected at all. The data suqqest that the constructed vectors are suitable for obtaining novel obese transqenic models based on a conditional genetic tissue ablation method.

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육상풍력 발전시설 지자체 규제 공간범위 산정 연구 (Estimating Spatial Scope of Local Government Ordinance for Onshore Wind Energy Generation Facilities)

  • 홍성희;김소라;박은정;이혜림;김진영;황수진;송정은
    • 신재생에너지
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    • 제18권4호
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    • pp.38-53
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    • 2022
  • T Recently, the government has been promoting the expansion and supply of renewable energy as an alternative for achieving carbon neutrality and the nationally determined contributions by 2030. In 2020, the Ministries of Industry and Environment and the Korea Forest Service collaborated to build a nationwide onshore wind energy siting atlas considering wind resources and forestry and environmental regulations focused on central regulations. In this study, the ordinances of the local governments were analyzed to examine the effects of regional location regulations on the expansion of onshore wind power energy generation facilities, in addition to those of central regulations. A development permit standard survey of 226 urban plan ordinances of the local governments nationwide showed that presently in 2022, 52 municipalities are applying regulations on wind energy generation facilities by ordinances. This is twice more than that in 2018, when renewable energy power generation facility development was difficult. Additionally, the location regulations applied by these ordinances were organized by items and regions, and regulatory characteristics, such as the number and scope, were analyzed by regions. To analyze the spatial distribution characteristics, JeollaNamdo was selected as the case area. A spatial DB was established for regulated areas based on the regional and central regulations, and the spatial distribution characteristics and the regulatory scope were compared and analyzed.