• Title/Summary/Keyword: 3' cis-elements

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Caffeic Acid Phenethyl Ester Induces the Expression of NAG-1 via Activating Transcription Factor 3 (ATF3를 통한 caffeic acid phenethyl ester에 의한 NAG-1 유전자의 발현 증가)

  • Park, Min-Hee;Chung, Chungwook;Lee, Seong Ho;Baek, Seung Joon;Kim, Jong Sik
    • Journal of Life Science
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    • v.28 no.1
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    • pp.37-42
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    • 2018
  • Non-steroidal anti-inflammatory drug-activated gene-1 (NAG-1) is a transforming growth factor beta (TGF-${\beta}$) superfamily gene associated with pro-apoptotic and anti-tumorigenic activities. In the present study, we investigated if caffeic acid phenethyl ester (CAPE) derived from propolis could induce the expression of anti-tumorigenic gene NAG-1. Our results indicate that CAPE significantly induced NAG-1 expression in a time- and concentration-dependent manner in HCT116 cells. We also found that CAPE induced NAG-1 expression in a concentration-dependent manner in another human colorectal cancer cell line, LOVO. In addition, CAPE triggered apoptosis, which was detected with Western blot analysis using poly-(ADP-ribose) polymerase antibody. NAG-1 induction by CAPE was not dependent on transcription factor p53, which was confirmed with Western blot analysis using p53 null HCT116 cells. The luciferase assay results indicated that the new cis-elements candidates were located between -474 and -1,086 of the NAG-1 gene promoter. CAPE dramatically induced activating transcription factor 3 (ATF3) expression, but not cAMP response element-binding protein (CREB), which shares the same binding sites with ATF3. The co-transfection experiment with pCG-ATF3 and pCREB showed that only ATF3 was associated with NAG-1 up-regulation by CAPE, whereas CREB had no effect. In conclusion, the results suggest that CAPE could induce the expression of anti-tumorigenic gene NAG-1 mainly through ATF3.

Functional Analysis of the Stress-Inducible Soybean Calmodulin Isoform-4 (GmCaM-4) Promoter in Transgenic Tobacco Plants

  • Park, Hyeong Cheol;Kim, Man Lyang;Kang, Yun Hwan;Jeong, Jae Cheol;Cheong, Mi Sun;Choi, Wonkyun;Lee, Sang Yeol;Cho, Moo Je;Kim, Min Chul;Chung, Woo Sik;Yun, Dae-Jin
    • Molecules and Cells
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    • v.27 no.4
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    • pp.475-480
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    • 2009
  • The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

AtbZIP16 and AtbZIP68, two new members of GBFs, can interact with other G group bZIPs in Arabidopsis thaliana

  • Shen, Huaishun;Cao, Kaiming;Wang, Xiping
    • BMB Reports
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    • v.41 no.2
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    • pp.132-138
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    • 2008
  • AtbZIP16 and AtbZIP68 are two putative G group bZIP transcription factors in Arabidopsis thaliana, the other three members of G group bZIPs are GBF1-3 which can bind G-box. Members of G group have conservative protein structure: highly homological basic region and a proline-rich domain in the N-terminal region. Here, we report that AtbZIP16 and AtbZIP68 could bind cis elements with ACGT core, such as G-box, Hex, C-box and As-1, but with different binding affinities which from high to low were G-box > Hex > C-box > As-1; AtbZIP16 and AtbZIP68 could form homodimer and form heterodimer with other members of G group; N-terminal proline rich domain of AtbZIP16 had transactivation activity in yeast cells while that of AtbZIP68 did not; AtbZIP16 and AtbZIP68 GFP fusion protein localized in the nucleus of onion epidermal cells. These results indicated that AtbZIP16 and AtbZIP68 were two new members of GBFs. In Arabidopsis, AtbZIP16 and AtbZIP68 may also participate in light-responsive process in which GBF1-3 are involved.

A Study on the dwellings of the Korean Diaspora of Uzbekistan (우즈베키스탄 거주 고려인의 주생호라에 관한 연구)

  • Lee YoungShim;Lee SangHae
    • Journal of the Korean Home Economics Association
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    • v.42 no.9
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    • pp.1-18
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    • 2004
  • Most of the Korean diaspora who lived in Yunhaeju moved to Central Asia in 1937 following the policy of deportation of Russia. Uzbekistan is the representative place for Korean diaspora to have settled down in Central Asia at that time and mort Korean diaspora in Central Asia are living in Uzbekistan now. Korean diaspora have maintained a traditional way of living for 140 years without a deep relationship with Korea. This study examined the dwellings of the Korean diaspora of Uzbekistan in Central Asia by visiting their houses and conducting interviews. Results of the research were as following: 1) The houses of the Korean diaspora in Uzbekistan in early times consisted of 2-3 bedrooms with Gudle. Gudle is the most traditional element of the Korean diaspora's house and it has been used as a place for gathering family members. 2) Korean diaspora's houses in Uzbekistan were built according to Russian and Uzbek style but most of the Korean diaspora's have a mixed style with traditional Korean elements of living.3) The changing process of planning and building codes of apartments in Uzbekistan is similar to that in other CIS nations and all of Russia. 4) Korean's food style is a mixture of Korean, Russian and Central Asian foods. Kimchi and Jang (bean paste) are essential for most Koreans and most Koreans are making these at home. 5) Most Koreans have maintained a good relationship with the Uzbek people and Koreans are very active in gelling along well with all. This enables them to exchange their culture eventually in various parts. This study could be the first step to supply basic information for the study of the Korean diaspora in Uzbekistan. Deeper research over a wider range is needed in Russia.

The Arabidopsis Phytocystatin AtCYS5 Enhances Seed Germination and Seedling Growth under Heat Stress Conditions

  • Song, Chieun;Kim, Taeyoon;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • v.40 no.8
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    • pp.577-586
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    • 2017
  • Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors that are implicated in protein turnover and stress responses. Here, we characterized a PhyCYS from Arabidopsis thaliana, which was designated AtCYS5. RT-qPCR analysis showed that the expression of AtCYS5 in germinating seeds was induced by heat stress (HS) and exogenous abscisic acid (ABA) treatment. Analysis of the expression of the ${\beta}-glucuronidase$ reporter gene under the control of the AtCYS5 promoter showed that AtCYS5 expression during seed germination was induced by HS and ABA. Constitutive overexpression of AtCYS5 driven by the cauliflower mosaic virus 35S promoter led to enhanced HS tolerance in transgenic Arabidopsis, which was characterized by higher fresh weight and root length compared to wild-type (WT) and knockout (cys5) plants grown under HS conditions. The HS tolerance of AtCYS5-overexpressing transgenic plants was associated with increased insensitivity to exogenous ABA during both seed germination and post-germination compared to WT and cys5. Although no HS elements were identified in the 5'-flanking region of AtCYS5, canonical ABA-responsive elements (ABREs) were detected. AtCYS5 was upregulated in ABAtreated protoplasts transiently co-expressing this gene and genes encoding bZIP ABRE-binding factors (ABFs and AREB3). In the absence of ABA, ABF1 and ABF3 directly bound to the ABREs in the AtCYS5 promoter, which activated the transcription of this gene in the presence of ABA. These results suggest that an ABA-dependent pathway plays a positive role in the HS-responsive expression of AtCYS5 during seed germination and post-germination growth.

Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Choristoneura fumiferana Granulovirus pk-1: A Baculoviral Protein Kinase

  • Giannopoulos, Paresa N.;Nassoury, Nasha;Lamontagne, Lucie;Guertin, Claude;Rashidan, Kianoush Khajeh
    • BMB Reports
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    • v.38 no.4
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    • pp.457-467
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    • 2005
  • Open reading frame (ORF) 3 on the Choristoneura fumiferana granulovirus (ChfuGV), located in the 11 kb fragment of the BamHI genomic bank encodes a predicted 32-kDa putative kinase protein. Bioinformatics analysis on the predicted amino acid sequence of ChfuGV PK-1 revealed the existence of 11 catalytic subdomains. Sequence analysis within the 5'-untranslated region (5'-UTR) of ChfuGV pk-1 indicates the presence of both putative early and late promoter motifs, indicating that pk-1 may be expressed throughout the infection cycle. Promoter sequence analysis reveals that pk-1 is deprived of a TATA box and appears instead to be regulated by other cis-acting transcriptional regulatory elements. Temporal transcription analysis by RT-PCR confirms the appearance of transcripts detected from 2 h p.i. until 72 h p.i. Northern blot hybridization characterizes pk-1 transcription as a 1.2 kb transcript. Homology comparisons reveal that ChfuGV PK-1 protein is most closely related to Phthorimaea operculalla granulovirus (PoGV) with 80% amino acid identity.

Versatile Luciferase Reporter Plasmids for Transcription Studies in Diverse Eukaryotic Cells (다양한 진핵생물 세포에서 전사 연구에 사용될 수 있는 Luciferase Reporter Plasmid의 개발)

  • 조영석;한동욱;백금희;박승필;윤상순;임운기;김정락;김한도;강호성
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.378-386
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    • 1996
  • We have developed a couple of new luciferase reporter plasmida with very low background reporter actlvltlea. One can be used to measure the promoter strength, after insertion of some promoter fragment into the reporter plasmid, and the other, with very low basal promoter actlvltlea, allis In studying eukaryotic transcriptional regulators. The latter reporter plasmid contains such cli elements as a 17 nucleotide long inftlator, Spl.blndIng sftes, GAL4 binding sltea, and bInding sitea for a certain Drosophila homeodomain proteins. In an attempt to construct an improved reporter plaimid by fadlltating transcriptional termination and minimizing any interference by cryptic promoters which may be preaent in the reporter pleamld DNA, we have inserted transsrlptional termination-related signals, a three tandem repeat of SV4O polyadenylatlon signal (AAA) and the putative transcrtptional termination signal (UMS) of the mouse c-mos gene, Into just upstream of the initIator, and the promoter actlvitiea were measured by a transIent expression assay employing the Drosophila Schneider line 2 cells. As expected, the basal promoter activitIes decreased maximally when both transcription termination related elements were inserted. Moreover, the reporter plasmld with the two elements allowed more sensitive measurement of transcriptional activation than the reporter piasmid without them. Theae reporter plasmids can be used for studying transcriptional regulators of higher organisms Including mammals as well as Droiophlla melanogaiter.

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Genetic and molecular analysis of the R-mb gene from maize (옥수수 R-mb 유전자의 유전분석과 그의 구조)

  • 윤필용;유삼규;송원용;윤충효;임용표
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.161-165
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    • 1997
  • The R-mb locus of maize is one of several genes that encode tissue-specific transcriptional regulator for the anthocyanin biosynthesis in plant parts and the aleurone layer in seeds. We found that the seed pigment frequencies gradually decreased at selfed progenies of the R-mb genetic stocks. In order to analyze the genomic structure of R-mb locus components, genomic Southern blot was performed by using R specific probe, pR-nj:1. Two bands were detected at the size of about 3.9 and 7.75kb. Five R-mb positive clones (mb-II, III, V,Ⅵ, and Ⅶ) were obtained by screening of maize genomic λFIXII library using R specific probe pR-nj:1. We constructed the restriction map of clone mb-II (7.75Kb positive) and mb-Ⅵ (3.9Kb positive), and have compared these with other R locus genes. From genetic and molecular analysis, it is suggested that R-mb complex consists two copy of R elements, and each element shows the paramutagenic and gene silencing effects by the fashion of cis-inactivation.

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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Shen, Xing-Jia;Yi, Yong-Zhu;Tang, Shun-Ming;Li, Yi-Ren;He, Jia-Lu;Wu, Xiang-Fu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.169-174
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    • 2004
  • Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (lue) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 - 2.0 $\mu\textrm{g}$/$m\ell$ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5-2.0 ${\mu}g$/$m\ell$ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.