• Title/Summary/Keyword: Transcription factor

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AUA as a Translation Initiation Site In Vitro for the Human Transcription Factor Sp3

  • Hernandez, Eric Moore;Johnson, Anna;Notario, Vicente;Chen, Andrew;Richert, John R.
    • BMB Reports
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    • v.35 no.3
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    • pp.273-282
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    • 2002
  • Sp3 is a bifunctional transcription factor that has been reported to stimulate or repress the transcription of numerous genes. Although the size of Sp3 mRNA is 4.0kb, the size of the known Sp3 cDNA sequence is 3.6kb. Thus, Sp3 functional studies have been performed with an artificially introduced start codon, and thus an amino-terminus that differs from the wild-type. Ideally, full-length cDNA expression vectors with the appropriate start codon should be utilized for these studies. Using 5'rapid amplification of cDNA ends, a full-length Sp3 cDNA clone was generated and the sequence verified in nine cell lines. No AUG initiation codon was present. However, stop codons were present in all three frames 5' to the known coding sequence. In vitro translation of this full-length cDNA clone produced the expected three isoforms-one at 100 kDa and two in the mid 60 kDa range. Electrophoretic mobility shift assays showed that the protein products had the ability to bind to the Sp1/3 consensus sequence. In vitro studies, using our Sp3 clone and site directed mutagenesis, identified the translation initiation site for the larger isoform as AUA. AUA has not been previously described as an endogenous initiation codon in eukaryotes.

Targeting Nrf2-Mediated Gene Transcription by Triterpenoids and Their Derivatives

  • Loboda, Agnieszka;Rojczyk-Golebiewska, Ewa;Bednarczyk-Cwynar, Barbara;Zaprutko, Lucjusz;Jozkowicz, Alicja;Dulak, Jozef
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.499-505
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    • 2012
  • Chemoprevention represents a strategy designed to protect cells or tissues against various carcinogens and carcinogenic metabolites derived from exogenous or endogenous sources. Recent studies indicate that plant-derived triterpenoids, like oleanolic acid, may exert cytoprotective functions via regulation of the activity of different transcription factors. The chemopreventive effects may be mediated through induction of the nuclear factor erythroid 2-related factor 2 (Nrf2) transcription factor. Activation of Nrf2 by triterpenoids induces the expression of phase 2 detoxifying and antioxidant enzymes such as NAD(P)H quinone oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1) - proteins which can protect cells or tissues against various toxic metabolites. On the other hand, inhibition of other transcription factors, like NF-${\kappa}B$ leads to the decrease in the pro-inflammatory gene expression. Moreover, the modulation of microRNAs activity may constitute a new mechanism responsible for valuable effects of triterpenoids. Recently, based on the structure of naturally occurring triterpenoids and with involvement of bioinformatics and computational chemistry, many synthetic analogs with improved biological properties have been obtained. Data from in vitro and in vivo experiments strongly suggest synthetic derivatives as promising candidates in the chemopreventive and chemotherapeutic strategies.

Inhibition of Adipogenesis in 3T3-L1 Adipocytes with Magnolia officinalis Extracts (후박 추출물의 지방세포 분화 억제 효능에 관한 연구)

  • Kim, Hyun-Ju;Lee, Yeo-Myeong;Kim, Yeon-Hyang;Won, Sun-Im;Choi, Sung-A;Choi, Shin-Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.2
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    • pp.117-123
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    • 2009
  • Magnolia extract, prepared from the Chinese herb Magnolia officinalis, is known for its potent anti-oxidative and anti-inflammatory effects. In this report, we showed that Magnolia extract inhibits adipocyte differentiation, as evidenced by reduced triglyceride (TG) accumulation. Also, Magnolia extract increased hormone sensitive lipase (HSL) protein level, and decreased the adipogenic transcription factor peroxisome proliferator activated receptor (PPAR)-${\gamma}$ protein and their corresponding mRNA. Our results suggest a potential apllication of Magnolia extract as anti-obesity agents inhibits adipocyte differentiation through the down-regulation of adipogenic transcription factors and other adipocyte-specific genes.

Inhibitory Efficacy of Angelica gigas Nakai on Microphthalmia-associated Transcription Factor (MITF), Tyrosinase Related Protein-1 (TRP-1), Tyrosinase Related Protein-2 (TRP-2), and Tyrosinase mRNA Expression in Melanoma Cells (B16F10) (멜라노마 세포에서 당귀추출물의 MITF, TRP-1, TRP-2, tyrosinase mRNA 발현 억제 효과)

  • Lee, Soo-Yeon;Lee, Jin-Young
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1336-1341
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    • 2013
  • The purpose of this study was to research the whitening effects of the extract from Angelica gigas Nakai, which is one of the most widely used herbal medicines in Asia. For whitening effects, the tyrosinase inhibition effect of the A. gigas Nakai extract was shown to be greater than 70% at 1,000 ${\mu}g/ml$ concentration. The result of measuring the cell toxicity effect of the extract from A. gigas Nakai on melanoma cells showed 99% toxicity at 500 ${\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase mRNA expression inhibitory effect by reverse transcription-PCR of the extract from A. gigas Nakai were decreased by 85.7%, 123.9%, 68.8%, and 208%, respectively, at 50 ${\mu}g/ml$ concentration. All these findings could verify that extract from A. gigas Nakai could have an effect on whitening. Moreover, extract from A. gigas Nakai has great potential as a cosmetic ingredient.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α

  • Kim, Jin-Ik;Kaufman, Randal J.;Back, Sung Hoon;Moon, Ja-Young
    • Molecules and Cells
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    • v.42 no.11
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    • pp.783-793
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    • 2019
  • When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.

Cucumber Mosaic Virus 1a Protein Interacts with the Tobacco SHE1 Transcription Factor and Partitions between the Nucleus and the Tonoplast Membrane

  • Yoon, Ju-Yeon;Palukaitis, Peter
    • The Plant Pathology Journal
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    • v.37 no.2
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    • pp.182-193
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    • 2021
  • The transcription factor SHE1 was identified as an interacting partner with the cucumber mosaic virus (CMV) 1a protein in the yeast two-hybrid system, by a pull-down assay, and via bimolecular fluorescent complementation. Using fluorescent-tagged proteins and confocal microscopy, the CMV 1a protein itself was found distributed predominantly between the nucleus and the tonoplast membrane, although it was also found in speckles in the cytoplasm. The SHE1 protein was localized in the nucleus, but in the presence of the CMV 1a protein was partitioned between the nucleus and the tonoplast membrane. SHE1 expression was induced by infection of tobacco with four tested viruses: CMV, tobacco mosaic virus, potato virus X and potato virus Y. Transgenic tobacco expressing the CMV 1a protein showed constitutive expression of SHE1, indicating that the CMV 1a protein may be responsible for its induction. However, previously, such plants also were shown to have less resistance to local and systemic movement of tobacco mosaic virus (TMV) expressing the green fluorescent protein, suggesting that the CMV 1a protein may act to prevent the function of the SHE1 protein. SHE1 is a member of the AP2/ERF class of transcription factors and is conserved in sequence in several Nicotiana species, although two clades of SHE1 could be discerned, including both different Nicotiana species and cultivars of tobacco, varying by the presence of particular insertions or deletions.

Expressional Evaluation of C/EBP Family, SREBP1, and Steroid Hormone Receptors in the Epididiymal Fat of Postnatally Developing Mouse

  • Lee, Yong-Seung;Lee, Ki-Ho
    • Development and Reproduction
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    • v.26 no.2
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    • pp.49-58
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    • 2022
  • The differentiation and development of preadipocyte into mature adipocyte are regulated by transcription factors, such as CCAAT enhancer binding protein (Cebp) gene family and sterol regulatory element binding transcription factor 1 (Srebp1). Steroid hormones give influences on the development and function of adipocyte. The present research examined expression patterns of CCAAT enhancer binding protein alpha (Cebpa), CCAAT enhancer binding protein beta (Cebpb), CCAAT enhancer binding protein gamma (Cebpg), sterol regulatory element binding transcription factor 1 (Srebp1), androgen receptor (Ar), and estrogen receptors (Esr) among different epididymal fat parts during postnatal period by quantitative real-time polymerase chain reaction. In the distal epididymal fat, expression of Cebpa, Cebpb, Cebpg, Srebp1, Ar, and Esr2 was increased until 12 months of age, while expression of Esr1 was decreased at 5 months of age and was not detectable after 8 months of age. In the proximal epididymal fat, transcript levels of Cebps and Srebp1 were increased at 8 months of age, followed by decreases of Cebpb and Cebpg transcript levels at 12 months of age. An additional increase of Srebp1 expression was observed at 12 months of age. Expression of Ar and Esr2 were increased until 8 months of age, followed by a drop of Ar expression level at 12 months of age. Expression pattern of Esr1 was similar to that in the distal epididymal fat. In the tail epididymal fat, expression of Cebpa, Cebpg, Srebp1, Ar, and Esr2 was increased with age. Esr1 was not detectable at all. The highest level of Cebpb was observed at 8 months of age. These data suggest the possibility of developmental and functional differentiation among the epididymal fat parts.

Whitening effect of novel peptide mixture by regulating melanosome biogenesis, transfer and degradation

  • Lee, Eung-Ji;Kim, Jandi;Jeong, Min Kyeong;Lee, Young Min;Chung, Yong Ji;Kim, Eun Mi
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.1
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    • pp.15-26
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    • 2021
  • Peptides are short chain of amino acids linked by peptide bonds. They are widely used as effective and biocompatible active ingredients in cosmetic industry. In this study, we developed novel peptide mixture and identified its anti-pigmentation effect on melanocytes and keratinocytes. Our results revealed that peptide mixture inhibited melanosome biogenesis through the regulation of microphthalmia-associated transcription factor, a key factor of melanogenesis in melanocytes. And we observed that peptide mixture inhibited melanosome uptake through the reduction of protease-activated receptor 2, a phagocytosis-related receptor in keratinocytes. Furthermore, peptide mixture activated autophagy system resulting in degradation of transferred melanosomes in keratinocytes. The anti-pigmentation effect of multi-targeting peptide mixture was assessed in a human skin equivalent model (MelanoDerm). Melanin contents in epidermal layer were significantly decreased by topical treatment of peptide mixture, suggesting that it can be applied as a novel cosmetics material having a whitening function.

Identification of Radiation-Sensitive Gene in U937 Cell by using cDNA-Chip Composed of Human Cancer Related Gene (U937 세포에서 발암관련 유전자들로 구성된 DNA chip을 이용한 방사선 감수성 유전자들의 선발)

  • 김종수;김인규;강경선;윤병수
    • Environmental Mutagens and Carcinogens
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    • v.22 no.1
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    • pp.54-59
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    • 2002
  • We have used cDNA microarray hybridization to identify gene regulated in response to gamma-irradiation in U-937 cell. The cDNA-chip was composed entirely of 1,000 human cancer related gene including apoptosis and angiogenesis etc. In gamma-irradiated U-937 cell, highly charged protein, ribosomal protein L32, four and a half LIM domains 3, lipocalin 2 (oncogene 24p3) and interleukin 15, ataxia telangiectasia mutated (includes complementation groups A, C and D) genes showed increased level of its transcription, and cell division cycle 25A, dihydrofolate reductase, topoisomerase (DNA) II beta(180kD), kinase suppressor of ras and strarigin genes showed reduced level of its transcription compared to untreated U-937 cell. The significant change of level of transcription was not found in well-known ionizing radiation(IR)-responsive gene, such as transcription factor TP53 and p53 related gene, except ataxia telangiectasia mutated gene.

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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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    • v.13 no.2
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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.