The Importance of REST for Development and Function of Beta Cells
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Transcription of Platelet-Derived Growth Factor Receptor a in Leydig Cells Involves Specificity Protein 1 and 3
125 Transcription of platelet-derived growth factor receptor a in Leydig cells involves specificity protein 1 and 3 Francis Bergeron1, Edward T Bagu1 and Jacques J Tremblay1,2 1Reproduction, Perinatal and Child Health, CHUQ Research Centre, CHUL Room T1-49, 2705 Laurier Boulevard, Que´bec, Que´bec, Canada G1V 4G2 2Department of Obstetrics and Gynecology, Faculty of Medicine, Centre for Research in Biology of Reproduction, Universite´ Laval, Que´bec, Que´bec, Canada G1V 0A6 (Correspondence should be addressed to J J Tremblay; Email: [email protected]) Abstract Platelet-derived growth factor (PDGF) A is secreted by Sertoli cells and acts on Leydig precursor cells, which express the receptor PDGFRA, triggering their differentiation into steroidogenically active Leydig cells. There is, however, no information regarding the molecular mechanisms that govern Pdgfra expression in Leydig cells. In this study, we isolated and characterized a 2.2 kb fragment of the rat Pdgfra 50-flanking sequence in the TM3 Leydig cell line, which endogenously expresses Pdgfra. A series of 50 progressive deletions of the Pdgfra promoter was generated and transfected in TM3 cells. Using this approach, two regions (K183/K154 and K154/K105), each conferring 46% of Pdgfra promoter activity, were identified. To better define the regulatory elements, trinucleotide mutations spanning the K154/K105 region were introduced by site-directed mutagenesis in the context of the K2.2kb Pdgfra promoter. Mutations that altered the TCCGAGGGAAAC sequence at K138 bp significantly decreased Pdgfra promoter activity in TM3 cells. Several proteins from TM3 nuclear extracts were found to bind to this G(C/A) motif in electromobility shift assay. -
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. -
1,25 Dihydroxyvitamin D-Mediated Orchestration of Anticancer
Kovalenko et al. BMC Genomics 2010, 11:26 http://www.biomedcentral.com/1471-2164/11/26 RESEARCH ARTICLE Open Access 1,25 dihydroxyvitamin D-mediated orchestration of anticancer, transcript-level effects in the immortalized, non-transformed prostate epithelial cell line, RWPE1 Pavlo L Kovalenko1, Zhentao Zhang1, Min Cui1, Steve K Clinton2, James C Fleet1* Abstract Background: Prostate cancer is the second leading cause of cancer mortality among US men. Epidemiological evidence suggests that high vitamin D status protects men from prostate cancer and the active form of vitamin D, 1a,25 dihydroxyvitamin D3 (1,25(OH)2D) has anti-cancer effects in cultured prostate cells. Still, the molecular mechanisms and the gene targets for vitamin D-mediated prostate cancer prevention are unknown. Results: We examined the effect of 1,25(OH)2D (+/- 100 nM, 6, 24, 48 h) on the transcript profile of proliferating RWPE1 cells, an immortalized, non-tumorigenic prostate epithelial cell line that is growth arrested by 1,25(OH)2D (Affymetrix U133 Plus 2.0, n = 4/treatment per time and dose). Our analysis revealed many transcript level changes at a 5% false detection rate: 6 h, 1571 (61% up), 24 h, 1816 (60% up), 48 h, 3566 (38% up). 288 transcripts were regulated similarly at all time points (182 up, 80 down) and many of the promoters for these transcripts contained putative vitamin D response elements. Functional analysis by pathway or Gene Set Analysis revealed early suppression of WNT, Notch, NF-kB, and IGF1 signaling. Transcripts related to inflammation were suppressed at 6 h (e.g. -
Association of Gene Ontology Categories with Decay Rate for Hepg2 Experiments These Tables Show Details for All Gene Ontology Categories
Supplementary Table 1: Association of Gene Ontology Categories with Decay Rate for HepG2 Experiments These tables show details for all Gene Ontology categories. Inferences for manual classification scheme shown at the bottom. Those categories used in Figure 1A are highlighted in bold. Standard Deviations are shown in parentheses. P-values less than 1E-20 are indicated with a "0". Rate r (hour^-1) Half-life < 2hr. Decay % GO Number Category Name Probe Sets Group Non-Group Distribution p-value In-Group Non-Group Representation p-value GO:0006350 transcription 1523 0.221 (0.009) 0.127 (0.002) FASTER 0 13.1 (0.4) 4.5 (0.1) OVER 0 GO:0006351 transcription, DNA-dependent 1498 0.220 (0.009) 0.127 (0.002) FASTER 0 13.0 (0.4) 4.5 (0.1) OVER 0 GO:0006355 regulation of transcription, DNA-dependent 1163 0.230 (0.011) 0.128 (0.002) FASTER 5.00E-21 14.2 (0.5) 4.6 (0.1) OVER 0 GO:0006366 transcription from Pol II promoter 845 0.225 (0.012) 0.130 (0.002) FASTER 1.88E-14 13.0 (0.5) 4.8 (0.1) OVER 0 GO:0006139 nucleobase, nucleoside, nucleotide and nucleic acid metabolism3004 0.173 (0.006) 0.127 (0.002) FASTER 1.28E-12 8.4 (0.2) 4.5 (0.1) OVER 0 GO:0006357 regulation of transcription from Pol II promoter 487 0.231 (0.016) 0.132 (0.002) FASTER 6.05E-10 13.5 (0.6) 4.9 (0.1) OVER 0 GO:0008283 cell proliferation 625 0.189 (0.014) 0.132 (0.002) FASTER 1.95E-05 10.1 (0.6) 5.0 (0.1) OVER 1.50E-20 GO:0006513 monoubiquitination 36 0.305 (0.049) 0.134 (0.002) FASTER 2.69E-04 25.4 (4.4) 5.1 (0.1) OVER 2.04E-06 GO:0007050 cell cycle arrest 57 0.311 (0.054) 0.133 (0.002) -
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. -
Functional Characterisation of MYT1L, a Brain-Specific Transcriptional Regulator
This electronic thesis or dissertation has been downloaded from the King’s Research Portal at https://kclpure.kcl.ac.uk/portal/ Functional characterisation of MYT1L, a brain-specific transcriptional regulator Kepa, Agnieszka Maria Awarding institution: King's College London The copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without proper acknowledgement. END USER LICENCE AGREEMENT Unless another licence is stated on the immediately following page this work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International licence. https://creativecommons.org/licenses/by-nc-nd/4.0/ You are free to copy, distribute and transmit the work Under the following conditions: Attribution: You must attribute the work in the manner specified by the author (but not in any way that suggests that they endorse you or your use of the work). Non Commercial: You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. Take down policy If you believe that this document breaches copyright please contact [email protected] providing details, and we will remove access to the work immediately and investigate your claim. Download date: 04. Oct. 2021 Functional characterisation of MYT1L, a brain-specific transcriptional regulator Agnieszka Kępa 2014 A thesis submitted to King’s College London for the degree of Doctor of Philosophy MRC Social, Genetic and Developmental Psychiatry Centre, Institute of Psychiatry ABSTRACT Abnormalities in brain development and maladaptive plasticity are thought to underlay a range of neurodevelopmental and neurological disabilities and disorders, such as autism spectrum disorders, schizophrenia and learning disability. -
(12) Patent Application Publication (10) Pub. No.: US 2006/0068395 A1 Wood Et Al
US 2006.0068395A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2006/0068395 A1 Wood et al. (43) Pub. Date: Mar. 30, 2006 (54) SYNTHETIC NUCLEIC ACID MOLECULE (21) Appl. No.: 10/943,508 COMPOSITIONS AND METHODS OF PREPARATION (22) Filed: Sep. 17, 2004 (76) Inventors: Keith V. Wood, Mt. Horeb, WI (US); Publication Classification Monika G. Wood, Mt. Horeb, WI (US); Brian Almond, Fitchburg, WI (51) Int. Cl. (US); Aileen Paguio, Madison, WI CI2O I/68 (2006.01) (US); Frank Fan, Madison, WI (US) C7H 2L/04 (2006.01) (52) U.S. Cl. ........................... 435/6: 435/320.1; 536/23.1 Correspondence Address: SCHWEGMAN, LUNDBERG, WOESSNER & (57) ABSTRACT KLUTH 1600 TCF TOWER A method to prepare synthetic nucleic acid molecules having 121 SOUTHEIGHT STREET reduced inappropriate or unintended transcriptional charac MINNEAPOLIS, MN 55402 (US) teristics when expressed in a particular host cell. Patent Application Publication Mar. 30, 2006 Sheet 1 of 2 US 2006/0068395 A1 Figure 1 Amino Acid Codon Phe UUU, UUC Ser UCU, UCC, UCA, UCG, AGU, AGC Tyr UAU, UAC Cys UGU, UGC Leu UUA, UUG, CUU, CUC, CUA, CUG Trp UGG Pro CCU, CCC, CCA, CCG His CAU, CAC Arg CGU, CGC, CGA, CGG, AGA, AGG Gln CAA, CAG Ile AUU, AUC, AUA Thr ACU, ACC, ACA, ACG ASn AAU, AAC LyS AAA, AAG Met AUG Val GUU, GUC, GUA, GUG Ala GCU, GCC, GCA, GCG Asp GAU, GAC Gly GGU, GGC, GGA, GGG Glu GAA, GAG Patent Application Publication Mar. 30, 2006 Sheet 2 of 2 US 2006/0068395 A1 Spd Sequence pGL4B-4NN3. -
BMC Developmental Biology Biomed Central
BMC Developmental Biology BioMed Central Research article Open Access Identification of known and novel pancreas genes expressed downstream of Nkx2.2 during development Keith R Anderson1, Peter White3,4, Klaus H Kaestner3 and Lori Sussel*1,2 Address: 1Department of Biochemistry and Program in Molecular Biology, University of Colorado Health Science Center, Denver, CO, 80045, USA, 2Department of Genetics and Development, Columbia University, New York, NY 10032, USA, 3Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, 752B CRB, 415 Curie Blvd, Philadelphia, Pennsylvania 19104, USA and 4The Research Institute at Nationwide Children's Hospital, 700 Childrens Drive, Columbus, OH 43205, USA Email: Keith R Anderson - [email protected]; Peter White - [email protected]; Klaus H Kaestner - [email protected]; Lori Sussel* - [email protected] * Corresponding author Published: 10 December 2009 Received: 27 May 2009 Accepted: 10 December 2009 BMC Developmental Biology 2009, 9:65 doi:10.1186/1471-213X-9-65 This article is available from: http://www.biomedcentral.com/1471-213X/9/65 © 2009 Anderson et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: The homeodomain containing transcription factor Nkx2.2 is essential for the differentiation of pancreatic endocrine cells. Deletion of Nkx2.2 in mice leads to misspecification of islet cell types; insulin-expressing β cells and glucagon-expressing α cells are replaced by ghrelin- expressing cells. -
Upregulation of Myt1 Promotes Acquired Resistance of Cancer Cells to Wee1 Inhibition Cody W
Published OnlineFirst October 8, 2019; DOI: 10.1158/0008-5472.CAN-19-1961 Cancer Molecular Cell Biology Research Upregulation of Myt1 Promotes Acquired Resistance of Cancer Cells to Wee1 Inhibition Cody W. Lewis1,2,3, Amirali B. Bukhari1,2,3, Edric J. Xiao1,3, Won-Shik Choi1,2,3, Joanne D. Smith1,2,3, Ellen Homola4, John R. Mackey1,5, Shelagh D. Campbell4, Armin M. Gamper1,2,3, and Gordon K. Chan1,2,3 Abstract Adavosertib (also known as AZD1775 or MK1775) is a tion of Cdk1 induced aberrant mitosis and cell death by small-molecule inhibitor of the protein kinase Wee1, with mitotic catastrophe. Cancer cells with intrinsic adavosertib single-agent activity in multiple solid tumors, including sar- resistance had higher levels of Myt1 compared with sensitive coma, glioblastoma, and head and neck cancer. Adavosertib cells. Furthermore, cancer cells that acquired resistance fol- also shows promising results in combination with genotoxic lowing short-term adavosertib treatment had higher levels of agents such as ionizing radiation or chemotherapy. Previous Myt1 compared with mock-treated cells. Downregulating studies have investigated molecular mechanisms of primary Myt1 enhanced ectopic Cdk1 activity and restored sensitivity resistance to Wee1 inhibition. Here, we investigated mechan- to adavosertib. These data demonstrate that upregulating Myt1 isms of acquired resistance to Wee1 inhibition, focusing on the is a mechanism by which cancer cells acquire resistance to role of the Wee1-related kinase Myt1. Myt1 and Wee1 kinases adavosertib. were both capable of phosphorylating and inhibiting Cdk1/ cyclin B, the key enzymatic complex required for mitosis, Significance: Myt1 is a candidate predictive biomarker of demonstrating their functional redundancy. -
MYT1 Protein MYT1 Protein
Catalogue # Aliquot Size M67-34G-20 20 µg M67-34G-50 50 µg MYT1 Protein Full length recombinant protein expressed in Sf9 cells Catalog # M67-34G Lot # N353 -3 Product Description Purity Recombinant full-length human MYT1 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. The gene accession number is NM_004203 . The purity was determined to be Gene Aliases >80% by densitometry. Approx. MW 93 kDa . PKMYT1, FLJ20093, DKFZp547K1610 Formulation Recombinant protein stored in 50mM Tris-HCl, pH 7.5, 50mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM DTT, 0.1mM PMSF, 25% glycerol. Storage and Stability Store product at –70 oC. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles. Scientific Background MYT1 is a member of a family of neural specific, zinc finger-containing DNA-binding proteins which binds to the promoter regions of proteolipid proteins of the central nervous system and plays a role in the developing nervous system. MYT1 of the oligodendrocyte lineage, along with a closely related CCHC zinc finger, is MYT1 Protein expressed in developing neurons in the mammalian Full length recombinant protein expressed in Sf9 cells central nervous system(1). The recombinant MYT1 fragments containing either the upstream 2 zinc fingers or Catalog Number M67-34G the downstream 4 zinc fingers bound the same cis Specific Lot Number N353-3 regulatory element in the PLP1 promoter in vitro and DNA Purity >80% binding required Zn(2+), but not other divalent cations Concentration 0.1 µ g/ µl Stability 1yr At –70 oC from date of shipment (2). -
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. -
Myt1l Safeguards Neuronal Identity by Actively Repressing Many Non-Neuronal Fates Moritz Mall1, Michael S
LETTER doi:10.1038/nature21722 Myt1l safeguards neuronal identity by actively repressing many non-neuronal fates Moritz Mall1, Michael S. Kareta1†, Soham Chanda1,2, Henrik Ahlenius3, Nicholas Perotti1, Bo Zhou1,2, Sarah D. Grieder1, Xuecai Ge4†, Sienna Drake3, Cheen Euong Ang1, Brandon M. Walker1, Thomas Vierbuchen1†, Daniel R. Fuentes1, Philip Brennecke5†, Kazuhiro R. Nitta6†, Arttu Jolma6, Lars M. Steinmetz5,7, Jussi Taipale6,8, Thomas C. Südhof2 & Marius Wernig1 Normal differentiation and induced reprogramming require human Myt1l (Extended Data Fig. 1). Chromatin immunoprecipita- the activation of target cell programs and silencing of donor cell tion followed by DNA sequencing (ChIP–seq) of endogenous Myt1l programs1,2. In reprogramming, the same factors are often used to in fetal neurons (embryonic day (E) 13.5) and ectopic Myt1l in mouse reprogram many different donor cell types3. As most developmental embryonic fibroblasts (MEFs) two days after induction identified 3,325 repressors, such as RE1-silencing transcription factor (REST) and high-confidence Myt1l peaks that overlapped remarkably well between Groucho (also known as TLE), are considered lineage-specific neurons and MEFs (Fig. 1a, Extended Data Fig. 2, Supplementary repressors4,5, it remains unclear how identical combinations of Table 1). Thus, similar to the pioneer factor Ascl1, Myt1l can access transcription factors can silence so many different donor programs. the majority of its cognate DNA binding sites even in a distantly related Distinct lineage repressors would have to be induced in different cell type. However, unlike Ascl1 targets8, the chromatin at Myt1l donor cell types. Here, by studying the reprogramming of mouse fibroblasts to neurons, we found that the pan neuron-specific a Myt1l Myt1l b Myt1l Ascl1 Random Myt1l 6 Ascl1 + Brn2 endogenous transcription factor Myt1-like (Myt1l) exerts its pro-neuronal Closed 0.030 function by direct repression of many different somatic lineage k programs except the neuronal program.