• 제목/요약/키워드: Transcriptional activation domain

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PI3-Kinase and PDK-1 Regulate HDAC1-mediated Transcriptional Repression of Transcription Factor NF-κB

  • Choi, Yong Seok;Jeong, Sunjoo
    • Molecules and Cells
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    • 제20권2호
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    • pp.241-246
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    • 2005
  • PDK-1 activates PI3-kinase/Akt signaling and regulates fundamental cellular functions, such as growth and survival. NF-${\kappa}B$ is involved in the induction of a variety of cellular genes affecting immunity, inflammation and the resistance to apoptosis induced by some anti-cancer drugs. Even though the crucial involvement of the PI3-kinase/Akt pathway in the anti-apoptotic activation of NF-${\kappa}B$ is well known, the exact role of PDK-1 as well as PI3-kinase/Akt in NF-vactivation is not understood. Here we demonstrate that PDK-1 plays a pivotal role in transcriptional activation of NF-${\kappa}B$ by dissociating the transcriptional co-repressor HDAC1 from the p65 subunit of NF-${\kappa}B$. The association of CBP with p65 was not directly modulated by PDK-1 or by PI3-kinase. Etoposide activated NF-${\kappa}B$ through PI3-kinase/Akt, and the transcription activation domain (TAD) of p65 was further activated by wild-type PDK-1. Overexpression of a dominant negative PDK-1 mutant decreased etoposide-induced NF-${\kappa}B$ transcription and further down-regulated the ectopic HDAC1-mediated decrease in NF-${\kappa}B$ transcriptional activity. Thus activation of PDK-1 relieves the HDAC1-mediated repression of NF-${\kappa}B$ that may be related to basal as well as activated transcription by NF-${\kappa}B$. This effect may also explain the role of the PI3-kinase/PDK-1 pathway in the anti-apoptotic function of NF-${\kappa}B$ associated with the chemoresistance of cancer cells.

Overexpression of OsNAC17 enhances drought tolerance in rice

  • Kim, Tae Hwan;Kim, Ju-Kon
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.168-168
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    • 2017
  • Drought conditions during cultivation reduce agricultural production yield less than a theoretical maximum yield under normal condition. Plant specific NAC transcription factors in rice are known to play an essential roles in stress resistance transcriptional regulation. In this study, we report the rice (Oryza sativa L japonica) NAM, AFTF and CUC transcription factor OsNAC17, which is predominantly induced by abiotic stress in leaf, was contribute to the drought tolerance mediated reactive oxygen species (ROS) in transgenic rice plants. Constitutive (PGD1) promoter was introduced to overexpress OsNAC17 and produced the transgenic PDG1:OsNAC17. Overexpression of OsNAC17 throughout the whole plant improved drought resistance phenotype at the vegetative stage. Morphological characteristics such as grain yield, grain filling rate, and total grain weight improved by 22~64% over wild type plants under drought conditions during the reproductive stage. The improved drought tolerance in transgenic rice was involved in reducing stomatal density up to 15% than in wild type plants and in increasing reactive oxygen species-scavenging enzyme. DEG profiling experiment identified 119 up-regulated genes by more than twofold (P<0.01). These genes included UDP-glycosyltransferase family protein, similar to 2-alkenal reductase (NADPH-dependent oxireductase), similar to retinol dehydrogenase 12, Lipoxygenase, and NB-ARC domain containing protein related in cell death. Furthermore, OsNAC17 was act as a transcriptional activator, which has an activation domain in C-terminal region. These result demonstrate that the overexpression of OsNAC17 improve drought tolerance by regulating ROS scavenging enzymes and by reducing stomatal density

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Phosphorylation Status of RNA Polymerase II Carboxyl-terminal Domain in Porcine Oocytes and Early Embryos

  • Oqani, Reza K.;Zhang, Jin Yu;Lee, Min-Gu;Diao, Yun Fei;Jin, Dong-Il
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권6호
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    • pp.789-793
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    • 2012
  • Fertilization of the oocyte commences embryogenesis during which maternally inherited mRNAs are degraded and the embryonic genome is activated. Transcription of embryonic mRNA is initiated by embryonic genome activation (EGA). RNA polymerase II (RNA Pol II) is responsible for the synthesis of mRNAs and most small nuclear RNAs, and consists of 12 subunits, the largest of which characteristically harbors a unique C-terminal domain (CTD). Transcriptional activity of RNA Pol II is highly regulated, in particular, by phosphorylation of serine residues in the CTD. Here, we have shown the presence of RNA Pol II CTD phosphoisoforms in porcine oocytes and preimplantation embryos. The distribution pattern as well as phosphorylation dynamics in germinal vesicles and during embryogenesis differed in developmental stages with these isoforms, indicating a role of RNA Pol II CTD phosphorylation at the serine residue in transcriptional activation during both oocyte growth and embryonic genome activation. We additionally examined the effects of the RNA Pol II inhibitor, ${\alpha}$-amanitin, on embryo development. Our results show that inhibition of polymerase, even at very early stages and for a short period of time, dramatically impaired blastocyst formation. These findings collectively suggest that the functionality of maternal RNA Pol II, and consequently, expression of early genes regulated by this enzyme are essential for proper embryo development.

Molecular cloning of metal-responsive transcription factor-1 (MTF-1) and transcriptional responses to metal and heat stresses in Pacific abalone, Haliotis discus hannai

  • Lee, Sang Yoon;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • 제20권7호
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    • pp.9.1-9.13
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    • 2017
  • Background: Metal-responsive transcription factor-1 (MTF-1) is a key transcriptional regulator playing crucial roles in metal homeostasis and cellular adaptation to diverse oxidative stresses. In order to understand cellular pathways associated with metal regulation and stress responses in Pacific abalone (Haliotis discus hannai), this study was aimed to isolate the genetic determinant of abalone MTF-1 and to examine its expression characteristics under basal and experimentally stimulated conditions. Results: The abalone MTF-1 shared conserved features in zinc-finger DNA binding domain with its orthologs; however, it represented a non-conservative shape in presumed transactivation domain region with the lack of typical motifs for nuclear export signal (NES) and Cys-cluster. Abalone MTF-1 promoter exhibited various transcription factor binding motifs that would be potentially related with metal regulation, stress responses, and development. The highest messenger RNA (mRNA) expression level of MTF-1 was observed in the testes, and MTF-1 transcripts were detected during the entire period of embryonic and early ontogenic developments. Abalone MTF-1 was found to be Cd inducible and highly modulated by heat shock treatment. Conclusion: Abalone MTF-1 possesses a non-consensus structure of activation domains and represents distinct features for its activation mechanism in response to metal overload and heat stress. The activation mechanism of abalone MTF-1 might include both indirect zinc sensing and direct de novo synthesis of transcripts. Taken together, results from this study could be a useful basis for future researches on stress physiology of this abalone species, particularly with regard to heavy metal detoxification and thermal adaptation.

Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain

  • Itoh, Ken;Wakabayashi, Nobunao;Katoh, Yasutake;Ishii, Tetsuro;Igarashi, Kazuhiko;Engel, James Douglas;Yamamoto, Masayuki
    • 한국환경성돌연변이발암원학회:학술대회논문집
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    • 한국환경성돌연변이발암원학회 2002년도 Current Trends in Toxicological Sciences
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    • pp.25-35
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    • 2002
  • Transcription factor Nrf2 is essential for the antioxidant responsive element (ARE)-mediated induction of phase II detoxifying and oxidative stress enzyme genes. Detailed analysis of differential Nrf2 activity displayed in transfected cell lines ultimately led to the identification of a new protein, which we named Keap1, that suppresses Nrf2 transcriptional activity by specific binding to its evolutionarily conserved amino-terminal regulatory domain. The closest homolog of Keap1 is a Drosophila actin-binding protein called Kelch, implying that Keap1 might be a Nrf2 cytoplasmic effector. We then showed that electrophilic agents antagonize Keap1 inhibition of Nrf2 activity in vivo, allowing Nrf2 to traverse from the cytoplasm to the nucleus and potentiate the ARE response. We postulate that Keap1 and Nrf2 constitute a crucial cellular sensor for oxidative stress, and together mediate a key step in the signaling pathway that leads to transcriptional activation by this novel Nrf2 nuclear shuttling mechanism. The activation of Nrf2 leads in turn to the induction of phase II enzyme and antioxidative stress genes in response to electrophiles and reactive oxygen species.

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An NMR study on the intrinsically disordered core transactivation domain of human glucocorticoid receptor

  • Kim, Do-Hyoung;Wright, Anthony;Han, Kyou-Hoon
    • BMB Reports
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    • 제50권10호
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    • pp.522-527
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    • 2017
  • A large number of transcriptional activation domains (TADs) are intrinsically unstructured, meaning they are devoid of a three-dimensional structure. The fact that these TADs are transcriptionally active without forming a 3-D structure raises the question of what features in these domains enable them to function. One of two TADs in human glucocorticoid receptor (hGR) is located at its N-terminus and is responsible for ~70% of the transcriptional activity of hGR. This 58-residue intrinsically-disordered TAD, named tau1c in an earlier study, was shown to form three helices under trifluoroethanol, which might be important for its activity. We carried out heteronuclear multi-dimensional NMR experiments on hGR tau1c in a more physiological aqueous buffer solution and found that it forms three helices that are ~30% pre-populated. Since pre-populated helices in several TADs were shown to be key elements for transcriptional activity, the three pre-formed helices in hGR tau1c delineated in this study should be critical determinants of the transcriptional activity of hGR. The presence of pre-structured helices in hGR tau1c strongly suggests that the existence of pre-structured motifs in target-unbound TADs is a very broad phenomenon.

Heterologous Expression of Streptomyces albus Genes Linked to an Integrating Element and Activation of Antibiotic Production

  • Kwon, Hyung-Jin;Lee, Soon-Youl;Hong, Soon-Kwang;Park, Uhn-Mee;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.488-497
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    • 1999
  • Probing Streptomyces albus ATCC 21838 chromosomal DNA with a proline tRNA sequence resulted in an isolation of a putative integrating element in the 6.4-kb EcoRI fragment. It was found that Streptomyces lividans TK-24 transformed with a cloned DNA fragment on a multicopy plasmid, produced a higher level of spore pigment and mycelial red pigment on a regeneration agar. Furthermore, the transformant S. lividans TK-24 produced a markedly increased level of undecylprodigiosin in a broth culture. A nucleotide sequence analysis of the cloned region revealed several open reading frames homologous to the integrases of integrating plasmids or temperate bacteriophages, signal-transducing regulatory proteins with a conserved ATP-binding domain, oxidoreductases ($\beta$-ketoacyl reductase), and an AraC-like transcriptional regulator. To examine the effect on antibiotic production, each coding region was overexpressed separately from the other genes in the region in S. lividans TK-24 with; pJHS3044 for the expression of the signal-transducing regulatory protein homologue, pJHS3045 for the homologue of oxidoreductase, and pJHS3051 for the homologue of the AraC-like transcriptional regulator. Phenotypic studies of S. lividans TK-24 strains harboring plasmids for the overexpression of individual genes suggested the following effects of the genes on antibiotic production: The oxidoreductase homologue stimulated the production of actinorhodin and undecylprodigiosin, which was influenced by the culture conditions; the homologue of the AraC-like transcriptional regulator was the most effective factor in antibiotic production within all the culture conditions tested; the signal-transducing regulatory protein homologue repressed the effect due to the homologue of the AraC-like transcriptional regulator, however, the antibiotic production was derepressed upon entering the stationary phase.

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Locus Control Region의 구조와 기능 (The Structure and Function of Locus Control Region)

  • 김애리
    • 생명과학회지
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    • 제17권11호
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    • pp.1587-1592
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    • 2007
  • Locus control region (LCR) is a cia-acting element which regulates the transcription of genes in developmental stage and/or tissue-specific pattern. Typically, LCR consists of several DNase I hypersensitive sites (HSs), where the binding motifs for transcriptional activators are present. The binding of activators to the HSs recruits chromatin modifying complexes to the LCR, opening chromatin structure and modifying histones covalently through the locus. LCR forms close physical contact with target gene located at a distance by looping away intervening region. In addition, non-coding RNA is transcribed from LCR toward target genes in continuously acetylated active domain. These structural and functional features of LCR suggest that the LCR plays many roles in chromatin activation and transcriptional regulation.

Transcriptional Activity of an Estrogen Receptor β Subtype in the Medaka Oryzias dancena

  • Maeng, Sejung;Yoon, Sung Woo;Kim, Eun Jeong;Nam, Yoon Kwon;Sohn, Young Chang
    • 한국발생생물학회지:발생과생식
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    • 제23권4호
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    • pp.333-344
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    • 2019
  • In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.