DOCSLIB.ORG

The Mammalian Mediator Complex

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Mammalian Mediator Complex View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector FEBS Letters 579 (2005) 904–908 FEBS 29061 Minireview The mammalian Mediator complex Joan Weliky Conawaya,b,c,*, Laurence Florensa, Shigeo Satoa, Chieri Tomomori-Satoa, Tari J. Parmelya, Tingting Yaoa, Selene K. Swansona, Charles A.S. Banksa, Michael P. Washburna, Ronald C. Conawaya,b a Stowers Institute for Medical Research, Kansas City, MO 64110, USA b Department of Biochemistry and Molecular Biology, Kansas University Medical Center, Kansas City, KS 66160, USA c Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA Received 25 October 2004; accepted 2 November 2004 Available online 24 November 2004 Edited by Gunnar von Heijne and Anders Liljas expressed in eukaryotes from yeast to man, is composed of Abstract The multiprotein Mediator (Med) complex is an evo- lutionarily conserved transcriptional regulator that plays impor- more than twenty subunits and has been named Mediator tant roles in activation and repression of RNA polymerase II (Med) for its role in mediating transcriptional signals from transcription. Prior studies identified a set of more than twenty DNA binding transcription factors bound at upstream pro- distinct polypeptides that compose the Saccharomyces cerevisiae moter elements and enhancers to RNA polymerase II and Mediator. Here we discuss efforts to characterize the subunit the general initiation factors bound at the core promoter sur- composition and associated activities of the mammalian Med rounding the transcriptional start site. complex. Ó 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. 2. Saccharomyces cerevisiae Mediator Keywords: Mass spectrometry; Mediator; Messenger RNA synthesis; MudPIT; RNA polymerase II; Transcription factor The Mediator was identified and first purified to near homo- geneity from S. cerevisiae by Kornberg and coworkers, by vir- tue of its ability to promote activator-dependent transcription by purified RNA polymerase II and the general initiation fac- tors in vitro [1]. This brought to light, for the first time, the 1. Introduction enormous complexity of the S. cerevisiae Med complex, by demonstrating that it was composed of some 20 proteins The initiation of messenger RNA synthesis is a major site for including the products of the Srb2, Srb4, Srb5, Srb6, and the regulation of gene expression. The initiation of eukaryotic Gal11 genes. The S. cerevisiae Srb genes were initially isolated messenger RNA synthesis is an elaborate biochemical process in a genetic screen for extragenic suppressors of an RNA poly- that is catalyzed by the 12 subunit enzyme RNA polymerase II merase II mutant containing a partial deletion of the heptad and requires, for simply a basal level of initiation, a minimum repeats in the carboxyl terminal domain (CTD) of its largest of five general initiation factors designated TFIIB, TFIID, subunit [2] and found to copurify in a high molecular mass TFIIE, TFIIF, and TFIIH, which comprise more than 20 dis- complex that was capable of supporting activator-dependent tinct polypeptides. Regulation of transcription initiation by RNA polymerase II transcription in vitro [3,4]. The Gal11 gene RNA polymerase II is further complicated by the requirement was initially isolated as gene required for transcription of for a very large, multisubunit ‘‘adaptor’’ that bridges RNA galactose-inducible genes [5]. polymerase II and its myriad DNA binding regulatory proteins Further characterization of the purified S. cerevisiae Med and transduces both positive and negative signals that turn on complex by Kornberg and coworkers led to the identification and off messenger RNA synthesis in response to the ever as Med subunits of the product of the Srb7 gene [6], the prod- changing microenvironment of the cell. This adaptor, which ucts of the Rgr1 and Sin4 genes [7], the product of the Rox3 is now known to be evolutionarily conserved and ubiquitously gene [8], the product of the Pgd1 Hrs1 gene [6], a collection of previously uncharacterized S. cerevisiae proteins, which they designated Med1, Med2, Med4, Med6, Med7, and Med8 [6], *Corresponding author. Fax: 816 926 2091. the products of the Cse2, Nut1, and Nut2 genes [9], and a pre- E-mail address: [email protected] (J.W. Conaway). viously uncharacterized protein, which they designated Med11 Abbreviations: ARC, activator-recruited cofactor; CRSP, cofactor re- [9]. Yeast strains carrying Rgr1 and Sin4 mutations exhibit quired for Sp1 activation; Cse2, chromosome segregation 2; CTD, similar transcriptional defects. Yeast strains carrying an Rgr1 carboxyl terminal domain; DRIP, vitamin D receptor-interacting pr- mutation failed to repress transcription of the Suc2 gene in otein; Med, Mediator; MudPIT, multidimensional protein identifica- the presence of glucose [10]. Yeast strains carrying a Sin4 tion technology; Nut, negative regulation of URS2; ORF, open reading frame; Rgr1, resistance to glucose repression 1; SMCC, Srb- mutation are defective in repressing transcription of the Gal1 Med-containing cofactor; Srb, suppressor of RNA polymerase B; and HO endonuclease genes [11,12]. Yeast strains carrying TRAP, thyroid hormone receptor-associated protein Rox3 mutations fail to repress transcription of the heme-regu- 0014-5793/$30.00 Ó 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. doi:10.1016/j.febslet.2004.11.031 J.W. Conaway et al. / FEBS Letters 579 (2005) 904–908 905 lated Cyc7 gene [13]. The Pdg1/Hrs1 gene was isolated in a ge- as TRIP2 (thyroid receptor interacting protein 2) and PBP netic screen for extragenic suppressors of the hyperrecombina- (PPAR binding protein) in yeast two-hybrid screens, by its tion phenotype of yeast carrying a deletion of the Hpr1 gene ability to interact in a ligand-dependent manner with the thy- [14]. Yeast strains carrying Pdg1/Hrs1 mutations were found roid hormone and PPAR receptors, respectively [37,38].In to be defective in activation of transcription of the Gal10 gene light of evidence that the MED1 protein is not required for [14]. The Nut1 and Nut2 genes were initially isolated in a genet- structural integrity of the mammalian Med complex and thus ic screen for yeast mutants defective in transcriptional activa- far appears to mediate activation of RNA polymerase II tran- tion by the DNA binding activator Swi4 [15]. The Cse2 gene scription only by nuclear receptors, the MED1 protein may was isolated in a genetic screen for yeast strains defective in play a specialized role in transcriptional regulation in mamma- chromosome segregation [16]. lian cells [39]. In addition to defining the subunit composition of the Despite significant similarities in the subunit compositions S. cerevisiae Med complex, Kornberg and coworkersÕ charac- of mammalian Med complexes isolated in different laborato- terization of the biochemical properties of their purified Med ries, apparent differences were notable (Fig. 1). The MED30 preparations led to the initial identification of many of the and MED31 proteins were initially identified only in the known Med-associated activities [1]. They and others pro- TRAP/SMCC complex [23,40]; the MED25 protein only in vided direct evidence that Med binds tightly not only to the the ARC, DRIP, and CRSP complexes [26–29]; the transcriptional activation domains of DNA binding transcrip- MED26 protein only in the CRSP and DRIP complexes tion factors [17,18], but also to RNA polymerase II to form a [27–29]; the MED8 protein only in the ARC and rat Med ‘‘holoenzyme’’ [1,4], consistent with the model that Med par- complexes [26]; the MED18 protein only in the mouse and ticipates directly in activator-dependent recruitment of poly- rat Med complexes [30,32]; and the MED9, MED11, merase to promoters. Although it is not yet completely MED19, MED22, MED28, and MED29 only in rat Med clear as to how Med interacts with RNA polymerase II, evi- [32–34]. dence suggests that yeast Med can bind directly to the heptad repeats in enzymeÕs CTD [1], and, through analysis of elec- tron micrographs of the holoenzyme, it appears that Med 4. Proteomic analysis of the mammalian Med complex makes multiple contacts with polymerase, with important contacts provided by Med interactions with the Rpb3 and Although the precise explanation for the apparent differ- Rpb11 polymerase subunits [19,20]. Perhaps not surprising ences in subunit compositions of the various mammalian in light of MedÕs significant interaction with RNA polymer- Med complexes characterized in different laboratories is cur- ase II in the preinitiation complex, yeast Med exerts potent rently not known, recent proteomic analyses suggests that stimulation of transcription initiation in reactions carried most of the Med-associated proteins identified in different lab- out in the absence of activators [1]. Binding of Med to oratories are bona fide subunits of the complex [41]. In these RNA polymerase II was found to stimulate phosphorylation experiments, multidimensional protein identification technol- of the CTD heptad repeats by the TFIIH-associated CTD ki- ogy (MudPIT) was used to characterize the subunit composi- nase [1]. Finally, S. cerevisiae Med was found to have associ- tion of a variety of mammalian Med complexes purified by ated histone acetyltransferase (HAT) activity likely carried anti-FLAG agarose immunoaffinity chromatography from out by its Nut1 subunit
Load more

Recommended publications
  • Genome-Wide Association Study of Body Weight
    Genome-wide association study of body weight in Australian Merino sheep reveals an orthologous region on OAR6 to human and bovine genomic regions affecting height and weight Hawlader A. Al-Mamun, Paul Kwan, Samuel A. Clark, Mohammad H. Ferdosi, Ross Tellam, Cedric Gondro To cite this version: Hawlader A. Al-Mamun, Paul Kwan, Samuel A. Clark, Mohammad H. Ferdosi, Ross Tellam, et al.. Genome-wide association study of body weight in Australian Merino sheep reveals an orthologous region on OAR6 to human and bovine genomic regions affecting height and weight. Genetics Selection Evolution, 2015, 47 (1), pp.66. 10.1186/s12711-015-0142-4. hal-01341302 HAL Id: hal-01341302 https://hal.archives-ouvertes.fr/hal-01341302 Submitted on 4 Jul 2016 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. et al. Genetics Selection Evolution Al-Mamun (2015) 47:66 Genetics DOI 10.1186/s12711-015-0142-4 Selection Evolution RESEARCH ARTICLE Open Access Genome-wide association study of body weight in Australian Merino sheep reveals an orthologous region on OAR6 to human and bovine genomic regions affecting height and weight Hawlader A. Al-Mamun1,2, Paul Kwan2, Samuel A.
    [Show full text]
  • Characterization of Genomic Copy Number Variation in Mus Musculus Associated with the Germline of Inbred and Wild Mouse Populations, Normal Development, and Cancer
    Western University Scholarship@Western Electronic Thesis and Dissertation Repository 4-18-2019 2:00 PM Characterization of genomic copy number variation in Mus musculus associated with the germline of inbred and wild mouse populations, normal development, and cancer Maja Milojevic The University of Western Ontario Supervisor Hill, Kathleen A. The University of Western Ontario Graduate Program in Biology A thesis submitted in partial fulfillment of the equirr ements for the degree in Doctor of Philosophy © Maja Milojevic 2019 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Genetics and Genomics Commons Recommended Citation Milojevic, Maja, "Characterization of genomic copy number variation in Mus musculus associated with the germline of inbred and wild mouse populations, normal development, and cancer" (2019). Electronic Thesis and Dissertation Repository. 6146. https://ir.lib.uwo.ca/etd/6146 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. Abstract Mus musculus is a human commensal species and an important model of human development and disease with a need for approaches to determine the contribution of copy number variants (CNVs) to genetic variation in laboratory and wild mice, and arising with normal mouse development and disease. Here, the Mouse Diversity Genotyping array (MDGA)-approach to CNV detection is developed to characterize CNV differences between laboratory and wild mice, between multiple normal tissues of the same mouse, and between primary mammary gland tumours and metastatic lung tissue.
    [Show full text]
  • Anti-MED30 (Aa 1-100) Polyclonal Antibody (CABT- BL5153) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use
    Anti-MED30 (aa 1-100) polyclonal antibody (CABT- BL5153) This product is for research use only and is not intended for diagnostic use. PRODUCT INFORMATION Product Overview Rabbit polyclonal antibody to Human MED30. Antigen Description The multiprotein TRAP/Mediator complex facilitates gene expression through a wide variety of transcriptional activators. MED30 is a component of this complex that appears to be metazoan specific (Baek et al., 2002 Immunogen Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human MED30 / TRAP25. Isotype IgG Source/Host Rabbit Species Reactivity Human Purification Immunogen affinity purified Conjugate Unconjugated Cellular Localization Nuclear Format Liquid Size 100 μg Buffer 1% BSA, PBS, pH 7.4 Preservative 0.02% Sodium Azide Storage Store at 4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze/thaw cycles. BACKGROUND Introduction The multiprotein TRAP/Mediator complex facilitates gene expression through a wide variety of transcriptional activators. MED30 is a component of this complex that appears to be metazoan specific (Baek et al., 2002 [PubMed 11909976]).[supplied by OMIM, Nov 2010] 45-1 Ramsey Road, Shirley, NY 11967, USA Email: [email protected] Tel: 1-631-624-4882 Fax: 1-631-938-8221 1 © Creative Diagnostics All Rights Reserved GENE INFORMATION Entrez Gene ID 90390 Protein Refseq NP_542382 UniProt ID Q96HR3 Chromosome Location 8q24.11 Pathway Developmental Biology, organism-specific biosystem; Gene Expression, organism-specific
    [Show full text]
  • Identification of Protein Complexes Associated with Myocardial Infarction Using a Bioinformatics Approach
    MOLECULAR MEDICINE REPORTS 18: 3569-3576, 2018 Identification of protein complexes associated with myocardial infarction using a bioinformatics approach NIANHUI JIAO1, YONGJIE QI1, CHANGLI LV2, HONGJUN LI3 and FENGYONG YANG1 1Intensive Care Unit; 2Emergency Department, Laiwu People's Hospital, Laiwu, Shandong 271199; 3Emergency Department, The Central Hospital of Tai'an, Tai'an, Shandong 271000, P.R. China Received November 3, 2016; Accepted January 3, 2018 DOI: 10.3892/mmr.2018.9414 Abstract. Myocardial infarction (MI) is a leading cause of clinical outcomes regarding high-risk MI. For example, muta- mortality and disability worldwide. Determination of the tions in the myocardial infarction-associated transcript have molecular mechanisms underlying the disease is crucial for been reported to cause susceptibility to MI (5). In addition, it identifying possible therapeutic targets and designing effective was demonstrated that mutations in the oxidized low-density treatments. On the basis that MI may be caused by dysfunc- lipoprotein receptor 1 gene may significantly increase the tional protein complexes rather than single genes, the present risk of MI (6). Although some MI-related genes have been study aimed to use a bioinformatics approach to identifying detected, many were identified independently and functional complexes that may serve important roles in the develop- associations among the genes have rarely been explored. ment of MI. By investigating the proteins involved in these Therefore, it is necessary to investigate MI from a systematic identified complexes, numerous proteins have been reported perspective, as the complex disease was reported to occur due that are related to MI, whereas other proteins interacted to the dysregulation of functional gene sets (7).
    [Show full text]
  • The Cyclin-Dependent Kinase 8 Module Sterically Blocks Mediator Interactions with RNA Polymerase II
    The cyclin-dependent kinase 8 module sterically blocks Mediator interactions with RNA polymerase II Hans Elmlund*†, Vera Baraznenok‡, Martin Lindahl†, Camilla O. Samuelsen§, Philip J. B. Koeck*¶, Steen Holmberg§, Hans Hebert*ʈ, and Claes M. Gustafsson‡ʈ *Department of Biosciences and Nutrition, Karolinska Institutet and School of Technology and Health, Royal Institute of Technology, Novum, SE-141 87 Huddinge, Sweden; †Department of Molecular Biophysics, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden; ‡Division of Metabolic Diseases, Karolinska Institutet, Novum, SE-141 86 Huddinge, Sweden; §Department of Genetics, Institute of Molecular Biology, Oester Farimagsgade 2A, DK-1353 Copenhagen K, Denmark; and ¶University College of Southern Stockholm, SE-141 57 Huddinge, Sweden Communicated by Roger D. Kornberg, Stanford University School of Medicine, Stanford, CA, August 28, 2006 (received for review February 21, 2006) CDK8 (cyclin-dependent kinase 8), along with CycC, Med12, and Here, we use the S. pombe system to investigate the molecular Med13, form a repressive module (the Cdk8 module) that prevents basis for the distinct functional properties of S and L Mediator. RNA polymerase II (pol II) interactions with Mediator. Here, we We find that the Cdk8 module binds to the pol II-binding cleft report that the ability of the Cdk8 module to prevent pol II of Mediator, where it sterically blocks interactions with the interactions is independent of the Cdk8-dependent kinase activity. polymerase. In contrast to earlier assumptions, the Cdk8 kinase We use electron microscopy and single-particle reconstruction to activity is dispensable for negative regulation of pol II interac- demonstrate that the Cdk8 module forms a distinct structural tions with Mediator.
    [Show full text]
  • Functional Diversification of the Nleg Effector Family in Enterohemorrhagic Escherichia Coli
    Functional diversification of the NleG effector family in enterohemorrhagic Escherichia coli Dylan Valleaua, Dustin J. Littleb, Dominika Borekc,d, Tatiana Skarinaa, Andrew T. Quailea, Rosa Di Leoa, Scott Houlistone,f, Alexander Lemake,f, Cheryl H. Arrowsmithe,f, Brian K. Coombesb, and Alexei Savchenkoa,g,1 aDepartment of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E5, Canada; bDepartment of Biochemistry and Biomedical Sciences, Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON L8S 4K1, Canada; cDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, TX 75390; dDepartment of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390; ePrincess Margaret Cancer Centre, University of Toronto, Toronto, ON M5G 2M9, Canada; fDepartment of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada; and gDepartment of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB T2N 4N1, Canada Edited by Ralph R. Isberg, Howard Hughes Medical Institute and Tufts University School of Medicine, Boston, MA, and approved August 15, 2018 (receivedfor review November 6, 2017) The pathogenic strategy of Escherichia coli and many other gram- chains. Depending on the length and nature of the polyubiquitin negative pathogens relies on the translocation of a specific set of chain, it posttranslationally regulates the target protein’s locali- proteins, called effectors, into the eukaryotic host cell during in- zation, activation, or degradation. Ubiquitination is a multistep fection. These effectors act in concert to modulate host cell pro- process which begins with the ubiquitin-activating enzyme (E1) cesses in favor of the invading pathogen. Injected by the type III using ATP to “charge” ubiquitin, covalently binding the ubiquitin secretion system (T3SS), the effector arsenal of enterohemorrhagic C terminus by a thioester linkage.
    [Show full text]
  • FBXL19 Recruits CDK-Mediator to Cpg Islands of Developmental Genes Priming Them for Activation During Lineage Commitment
    RESEARCH ARTICLE FBXL19 recruits CDK-Mediator to CpG islands of developmental genes priming them for activation during lineage commitment Emilia Dimitrova1, Takashi Kondo2†, Angelika Feldmann1†, Manabu Nakayama3, Yoko Koseki2, Rebecca Konietzny4‡, Benedikt M Kessler4, Haruhiko Koseki2,5, Robert J Klose1* 1Department of Biochemistry, University of Oxford, Oxford, United Kingdom; 2Laboratory for Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; 3Department of Technology Development, Kazusa DNA Research Institute, Kisarazu, Japan; 4Nuffield Department of Medicine, TDI Mass Spectrometry Laboratory, Target Discovery Institute, University of Oxford, Oxford, United Kingdom; 5CREST, Japan Science and Technology Agency, Kawaguchi, Japan Abstract CpG islands are gene regulatory elements associated with the majority of mammalian promoters, yet how they regulate gene expression remains poorly understood. Here, we identify *For correspondence: FBXL19 as a CpG island-binding protein in mouse embryonic stem (ES) cells and show that it [email protected] associates with the CDK-Mediator complex. We discover that FBXL19 recruits CDK-Mediator to †These authors contributed CpG island-associated promoters of non-transcribed developmental genes to prime these genes equally to this work for activation during cell lineage commitment. We further show that recognition of CpG islands by Present address: ‡Agilent FBXL19 is essential for mouse development. Together this reveals a new CpG island-centric Technologies, Waldbronn, mechanism for CDK-Mediator recruitment to developmental gene promoters in ES cells and a Germany requirement for CDK-Mediator in priming these developmental genes for activation during cell lineage commitment. Competing interests: The DOI: https://doi.org/10.7554/eLife.37084.001 authors declare that no competing interests exist.
    [Show full text]
  • Whole Genome Resequencing of Four Italian Sweet Pepper Landraces Provides Insights on Sequence Variation in Genes of Agronomic V
    www.nature.com/scientificreports OPEN Whole genome resequencing of four Italian sweet pepper landraces provides insights on sequence variation in genes of agronomic value Alberto Acquadro1, Lorenzo Barchi1 ✉ , Ezio Portis1, Mohamed Nourdine1, Cristiano Carli2, Simone Monge3, Danila Valentino1 & Sergio Lanteri1 Sweet pepper (Capsicum annuum L.) is a high value crop and one of the most widely grown vegetables belonging to the Solanaceae family. In addition to commercial varieties and F1 hybrids, a multitude of landraces are grown, whose genetic combination is the result of hundreds of years of random, environmental, and farmer selection. High genetic diversity exists in the landrace gene pool which however has scarcely been studied, thus bounding their cultivation. We re-sequenced four pepper inbred lines, within as many Italian landraces, which representative of as many fruit types: big sized blocky with sunken apex (‘Quadrato’) and protruding apex or heart shaped (‘Cuneo’), elongated (‘Corno’) and smaller sized sub-spherical (‘Tumaticot’). Each genomic sequence was obtained through Illumina platform at coverage ranging from 39 to 44×, and reconstructed at a chromosome scale. About 35.5k genes were predicted in each inbred line, of which 22,017 were shared among them and the reference genome (accession ‘CM334’). Distinctive variations in miRNAs, resistance gene analogues (RGAs) and susceptibility genes (S-genes) were detected. A detailed survey of the SNP/Indels occurring in genes afecting fruit size, shape and quality identifed the highest frequencies of variation in regulatory regions. Many structural variations were identifed as presence/absence variations (PAVs), notably in resistance gene analogues (RGAs) and in the capsanthin/capsorubin synthase (CCS) gene.
    [Show full text]
  • MED26 Antibody
    Product Datasheet MED26 Antibody Catalog No: #33633 Package Size: #33633-1 50ul #33633-2 100ul Orders: [email protected] Support: [email protected] Description Product Name MED26 Antibody Host Species Rabbit Clonality Polyclonal Purification The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. Applications WB Species Reactivity Hu Ms Specificity The antibody detects endogenous levels of total MED26 protein. Immunogen Type Peptide Immunogen Description Synthesized peptide derived from N-terminal of human MED26. Target Name MED26 Other Names ARC70; CRSP complex 7; CRSP70; MED26; activator-recruited cofactor 70 kDa component Accession No. Swiss-Prot: O95402NCBI Gene ID: 9441 SDS-PAGE MW 65kd Concentration 1.0mg/ml Formulation Rabbit IgG in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Storage Store at -20°C Application Details Western blotting: 1:500~1:3000 Images Western blot analysis of extracts from LOVO cells, using MED26 antibody #33633. Background Component of the Mediator complex, a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator Address: 8400 Baltimore Ave., Suite 302, College Park, MD 20740, USA http://www.sabbiotech.com 1 functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Ryu S., Nature 397:446-450(1999). Naeaer A.M., Nature 398:828-832(1999).
    [Show full text]
  • Cbx3 Maintains Lineage Specificity During Neural Differentiation
    Downloaded from genesdev.cshlp.org on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press RESEARCH COMMUNICATION et al. 2013). However, Cbx3 is also enriched at promoters Cbx3 maintains lineage of mouse embryonic fibroblast (MEF)-derived pre-iPSCs specificity during neural (preinduced pluripotent stem cells), where it does not cor- relate with H3K9 methylation (Sridharan et al. 2013). differentiation RNAi knockdown of Cbx3 promotes reprogramming of fi- broblasts to iPSCs (Sridharan et al. 2013). Chengyang Huang,1,2 Trent Su,1 Yong Xue,1 1 3 1 Cbx3 plays important roles in developmental processes Chen Cheng, Fides D. Lay, Robin A. McKee, (Morikawa et al. 2013). In model systems, Cbx3 promotes Meiyang Li,2 Ajay Vashisht,1 James Wohlschlegel,1 neuronal maturation, kidney development, differentia- Bennett G. Novitch,4 Kathrin Plath,1 tion of ESCs to smooth muscle in culture, and embryonic Siavash K. Kurdistani,1 and Michael Carey1 arteriogenesis (Xiao et al. 2011; Dihazi et al. 2015; Oshiro et al. 2015). Little is known of how Cbx3 functions in dif- 1Department of Biological Chemistry, Eli and Edythe Broad ferentiation, but its promoter localization suggested that Center for Regenerative Medicine and Stem Cell Research, David it might directly affect transcription through the preinitia- Geffen School of Medicine, University of California at Los tion complex (PIC) (Grunberg and Hahn 2013; Allen and Angeles, Los Angeles California 90095, USA; 2Department of Taatjes 2015). The PIC is assembled in response to activa- Neurobiology, Shantou University Medical College, Shantou tors and requires the Mediator coactivator complex to re- 515041, China; 3Department of Molecular, Cell, and cruit the general transcription factors and Pol II (Chen Developmental Biology, University of California at Los Angeles, et al.
    [Show full text]
  • Original Article Valproic Acid Decreases the Nuclear Localization of MDT-28, the Nematode Orthologue of MED28
    Original Article Valproic Acid Decreases the Nuclear Localization of MDT-28, the Nematode Orthologue of MED28 (Mediator complex / valproic acid / nuclear localization / MDT-28 / MED28) M. KOSTROUCHOVÁ1,2,*, V. KOSTROUCHOVÁ1, P. YILMA1, A. BENDA3, V. MANDYS2, M. KOSTROUCHOVÁ1,‡ 1Biocev, First Faculty of Medicine, Charles University, Prague, Czech Republic 2Department of Pathology, Third Faculty of Medicine, Charles University, Prague, Czech Republic 3Imaging Methods Core Facility, Biocev, Faculty of Science, Charles University, Prague, Czech Republic Abstract. Mediator is a multiprotein complex that tode orthologue of MED28, as a likely target of lysine connects regulation mediated by transcription fac- acetylation using bioinformatic prediction of post- tors with RNA polymerase II transcriptional machin- translational modifications. Lysine acetylation was ery and integrates signals from the cell regulatory experimentally confirmed using anti-acetyl lysine cascades with gene expression. One of the Mediator antibody on immunoprecipitated GFP::MDT-28 ex- subunits, Mediator complex subunit 28 (MED28), pressed in synchronized C. elegans. Valproic acid has a dual nuclear and cytoplasmic localization and (VPA), a known inhibitor of lysine deacetylases, en- function. In the nucleus, MED28 functions as part of hanced the lysine acetylation of GFP::MDT-28. At Mediator and in the cytoplasm, it interacts with cy- the subcellular level, VPA decreased the nuclear lo- toskeletal proteins and is part of the regulatory cas- calization of GFP::MDT-28 detected by fluorescence- cades including that of Grb2. MED28 thus has the lifetime imaging microscopy (FLIM). This indicates potential to bring cytoplasmic regulatory interac- that the nuclear pool of MDT-28 is regulated by a tions towards the centre of gene expression regula- mechanism sensitive to VPA and provides an indirect tion.
    [Show full text]
  • Diaezdbioinfo.Pdf
    TESIS DOCTORAL DISEÑO Y DESARROLLO DE UNA PLATAFORMA BIOINFORMÁTICA PARA LA INTEGRACIÓN, GESTIÓN Y VISUALIZACIÓN DE REDES DE INTERACCIÓN DE PROTEÍNAS E INTERACTOMAS DIEGO ALONSO LÓPEZ DIRECTORES DR. JAVIER DE LAS RIVAS SANZ DR. RODRIGO SANTAMARÍA VICENTE SALAMANCA, MARZO DE 2017 El Dr. Javier De Las Rivas Sanz, con D.N.I. 15949000H, Investigador Científico del Consejo Superior de Investigaciones Científicas (CSIC), director del grupo de Bioinformática y Genómica Funcional en el Instituto de Biología Molecular y Celular del Cáncer (CiC‐IBMCC), y profesor del Programa de Doctorado y del Máster de Biología y Clínica del Cáncer de dicho Instituto y la Universidad de Salamanca (USAL). Y el Dr. Rodrigo Santamaría Vicente, con D.N.I. 70879303L, Profesor Titular del Departamento de Informática y Automática de la Universidad de Salamanca (USAL), miembro del grupo de investigación VisUsal (Visualización de información y analítica visual) de dicha universidad y del grupo de investigación en Dinámica del Genoma y Epigenética del Instituto de Biología Funcional y Genómica (CSIC‐USAL). CERTIFICAN que han dirigido esta Tesis Doctoral titulada "DISEÑO Y DESARROLLO DE UNA PLATAFORMA BIOINFORMÁTICA PARA LA INTEGRACIÓN, GESTIÓN Y VISUALIZACIÓN DE REDES DE INTERACCIÓN DE PROTEÍNAS E INTERACTOMAS” realizada por D. Diego Alonso López, alumno del programa de doctorado de Ingeniería Informática de la Universidad de Salamanca. Y AUTORIZAN la presentación de la misma, considerando que reúne las condiciones de originalidad y contenidos requeridos para optar al grado de Doctor por la Universidad de Salamanca. En Salamanca, a 17 de marzo de 2017 Dr. Javier De Las Rivas Sanz Dr. Rodrigo Santamaría Vicente AGRADECIMIENTOS A mis directores de tesis, Javier De Las Rivas y Rodrigo Santamaría, gracias por darme la oportunidad de realizar este trabajo y facilitarme el camino con vuestros consejos y ayuda.
    [Show full text]