DOCSLIB.ORG

Glucocorticoid Receptor and Myocyte Enhancer Factor 2 Cooperate to Regulate the Expression of C-JUN in a Neuronal Context

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Glucocorticoid Receptor and Myocyte Enhancer Factor 2 Cooperate to Regulate the Expression of C-JUN in a Neuronal Context J Mol Neurosci (2012) 48:209–218 DOI 10.1007/s12031-012-9809-2 Glucocorticoid Receptor and Myocyte Enhancer Factor 2 Cooperate to Regulate the Expression of c-JUN in a Neuronal Context Niels Speksnijder & Kenneth V. Christensen & Michael Didriksen & E. Ronald De Kloet & Nicole A. Datson Received: 17 April 2012 /Accepted: 7 May 2012 /Published online: 24 May 2012 # The Author(s) 2012. This article is published with open access at Springerlink.com Abstract The glucocorticoid receptor (GR) and myocyte c-JUN. In conclusion, for the first time, we show that enhancer factor 2 (MEF2) are transcription factors involved activated GR requires MEF2 to regulate c-JUN. At the in neuronal plasticity. c-JUN, a target gene of GR and same time, GR influences MEF2 activity and DNA binding. MEF2, plays a role in regulating both synaptic strength These results give novel insight into the molecular interplay of and synapse number. The aim of this study was to investi- GR and MEF2 in the control of genes important for neuronal gate the nature of this dual regulation of c-JUN by GR and plasticity. MEF2 in a neuronal context. First, we showed that GR mediates the dexamethasone-induced suppression of c- Keywords Glucocorticoid receptor . Myocyte enhancer JUN mRNA expression. Next, we observed that GR activa- factor 2 . c-JUN . Dexamethasone . PC-12 cells tion resulted in an increase in phosphorylation of MEF2, a post-translational modification known to change MEF2 Abbreviations from a transcriptional enhancer to a repressor. In addition, CDK5 Cyclin-dependent kinase 5 we observed an enhanced binding of MEF2 to genomic sites ChIP Chromatin immuno-precipitation directly upstream of the c-JUN gene upon GR activation. DEX Dexamethasone Finally, in primary hippocampal neuronal cultures, knock- GBS GR-binding site down of MEF2 not only reduced c-JUN expression levels GR Glucocorticoid receptor but abolished GR regulation of c-JUN expression. This GRE Glucocorticoid response element suggests that MEF2 is necessary for GR regulation of HPA-axis Hypothalamic–pituitary–adrenal axis IEG Immediate-early gene MBS MEF2 binding site Electronic supplementary material The online version of this article (doi:10.1007/s12031-012-9809-2) contains supplementary material, MEF2 Myocyte enhancer factor 2 which is available to authorized users. PC-12 Pheochromocytoma cells N. Speksnijder : E. R. De Kloet : N. A. Datson (*) Division of Medical Pharmacology, Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University Medical Center, P.O. Box 9502, 2300 RA Leiden, The Netherlands Introduction e-mail: [email protected] : Neuronal plasticity, a change in the structure, function, and K. V. Christensen M. Didriksen organization of neurons in response to environmental stim- H. Lundbeck A/S R&D, 9 Ottiliavej, uli, underlies many key processes such as learning and DK-2500 Copenhagen-Valby, Denmark memory, adaptation, and behavioral sensitization. Changes in gene expression, governed by key transcription factors, N. A. Datson such as the glucocorticoid receptor (GR) and myocyte en- Department of Human Genetics, Leiden University Medical Center, hancer factor 2 (MEF2), underlie neuroplasticity. GR is Leiden, The Netherlands activated by glucocorticoid stress hormones, released by 210 J Mol Neurosci (2012) 48:209–218 the HPA-axis in response to stress. Upon activation, GR acts For GR blockade, cells were pretreated with VEH (0.1% as a ligand-activated transcription factor to influence expres- ethanol) or 1 mM RU486 (M8046, Sigma-Aldrich) for sion of a wide variety of genes, including genes involved in 60 min before addition of DEX or VEH. neuronal plasticity (Datson et al. 2008). MEF2 comprises a family of four members, MEF2a–d, showing distinct but Hippocampal Cultures partly overlapping expression patterns and is activated by neuronal activity. Upon activation, MEF2 regulates the ex- Newborn pups from NMRI mice were killed at postnatal pression of genes that control dendritic remodeling, result- day 1. Brains were isolated and kept in Hank’s balanced salt ing in the inhibition of synapse formation. Conversely, a solution on ice until dissection. Hippocampi were dissected decrease in MEF2 activity increases spine density (Flavell et in ice-cold dissection solution consisting of Krebs buffer al. 2006; Shalizi et al. 2006). supplemented with 3 mg/ml BSA, 1.2 mM MgSO4,and We previously showed that GR and MEF2 have several 2 mM HEPES. Hippocampi (n012) were transferred to a target genes in common, including the c-JUN gene (Datson conical tube containing 1.5 ml of dissection solution sup- et al. 2011a). c-JUN is a subunit of the transcription factor plemented with 184 μg/ml trypsin. The tissue was incubated AP-1 and is a ubiquitously expressed IEG with important at 37°C for 6 min. Subsequently, 3.5 ml of dissection solu- functions in cell death, differentiation, and inflammation tion supplemented with 0.65 mg/ml soyabean trypsin inhib- (Beck et al. 2009; Sun et al. 2005). The AP-1 family of itor; 10 μg/ml DNAse and 0.19 mM MgSO4 were added. transcription factors is recruited in the activation of neuronal The trypsinated and DNase-treated hippocampi were centri- circuits leading to long-term changes, such as long-term fuged at 100×g for 3 min. The supernatant was discarded, memory formation (Alberini 2009). MEF2 is known to and the conical part of the tube was filled with 1.5 ml of induce transcription of c-JUN (Kato et al. 1997; Aude- dissection solution supplemented with 5.2 mg/ml soyabean Garcia et al. 2010; Han and Prywes 1995), while GR trypsin inhibitor, 80 μg/ml DNAse, and 1.5 mM MgSO4. on the other hand is known to repress the expression of c-JUN The cells were dissociated by pipetting and left for 5 min at in vitro in AtT-20 cells and mouse fibroblast cells (Autelitano room temperature (RT), allowing remaining tissue to settle. 1994; Wei et al. 1998). The aim of this study was to The supernatant was transferred to a new tube containing investigate the molecular interplay of GR and MEF2 in a 3.5 ml of dissection solution supplemented with 132 μM neuronal context, using the shared target gene c-JUN as a CaCl2 and 120 μMMgSO4 and centrifuged for 10 min at proof-of-principle. 100×g. The cell pellet was resuspended in 3.5 ml of MEM II+ B27 (MEM buffer supplemented with 0.5% D-glucose, 0.22% bicarbonate, penicillin–streptomycin, 2 mM L-glutamate, Materials and Methods 10% NU-serum, and 2% B27). After resuspension, the concentrated cell solution was diluted to 7.5 ml MEM II+ Cell Culture and Treatment B27. Cells were plated at a density of 50,000 live cells/well in poly-D-lysine coated 96-well dishes. The yield from one pup Rat pheochromocytoma (PC-12) cells (passage # 15–29) (two hippocampi) was approximately 400,000 living cells. were cultured as described earlier (Morsink et al. 2006b). The day after plating, media was changed to MEM II+B27 In short, cells were grown in DMEM medium, supple- buffer supplemented with 1 μM AraC (cytosine arabinoside). mented with 0–10% fetal bovine serum and 0–10% horse The cells were left for 14 days in vitro before assaying. serum, dependent on the stage of neuronal differentiation. For mRNA and protein analysis cells were seeded at a Lentiviral shRNA Transduction, Stimulation, and RNA confluency of 30–50% in pre-coated six-well plates Purification (356400, BD Biosciences, San Jose, CA, USA). For ChIP experiments, the cells were seeded at 50% confluency in High titer batches (>5×10−8 TU/ml) of lentiviral particles pre-coated 175 cm2 plates (356478, BD Biosciences). Neu- harboring gene-specific short hairpin RNA (shRNA) target- ronal differentiation was achieved by giving 50 ng/ml ing MEF2A (Sigma, TRCN0000095959) as well lentiviral Nerve Growth Factor Beta (NGF-ß) (N2513, Sigma- particles harboring control shRNA (Sigma, SHC002V) were Aldrich, St. Louis, MO, USA) every other day for 10 days. purchased from Sigma. The day after plating, hippocampal Medium at day 9 of the differentiation was supplemented with cultures were transduced with lentiviral particles at the charcoal-stripped serum to deprive the medium of endoge- following concentrations: 150,000 lentiviral particles/well, nous steroids (Sarabdjitsingh et al. 2010b). At day 10, the cells 75,000 lentiviral particles/well, and 37,500 lentiviral particles/ were treated for 30, 60, 90, or 180 min, dependent on the well. At day 14, the hippocampal cultures were stimulated for experiment, with either vehicle (VEH) (0.1% ethanol) or 90 min with 100 nM dexamethasone diluted in astrocyte- 100 nM dexamethasone (DEX) (D1756, Sigma-Aldrich). conditioned media. The latter was to avoid glutamate-induced J Mol Neurosci (2012) 48:209–218 211 excitotoxicity by the media change. Subsequently, cells 1% NaDOC), and 2× with TE buffer (1 mM EDTA pH 8.0; were processed for RNA isolation using the Aurum Total 10 mM Tris–HCl pH 8.0). Subsequently, the DNA com- RNA 96 Kit (BioRad). plexes were eluted from the beads with 0.1 M NaHCO3 and 1% SDS, and the DNA was reverse-crosslinked o/n at 4°C in Real-Time Quantitative PCR 0.2 M NaCl. The samples were then treated for 1 h with RNAse at 37°C and the DNA purified using Nucleospin Total RNA was isolated using Trizol (15596, Invitrogen) columns. The DNA was eluted in TE buffer for RT-qPCR according to the manufacturer’s instructions. RNAwas diluted analysis. RT-qPCR on chromatin immunoprecipitation to 50 ng/μl, and cDNA was synthesized using the iScript (ChIP) material was performed directly on purified DNA. cDNA synthesis kit (170–8897, Bio-Rad, Hercules, CA, ChIP results were obtained by performing three individual USA) according to the manufacturer’s protocol.
Load more

Recommended publications
  • Mutant P53 Protects Cells from 12-O-Tetradecanoylphorbol-13- Acetate–Induced Death by Attenuatingactivating Transcription Factor 3 Induction
    Research Article Mutant p53 Protects Cells from 12-O-Tetradecanoylphorbol-13- Acetate–Induced Death by AttenuatingActivating Transcription Factor 3 Induction Yosef Buganim,1 Eyal Kalo,1 Ran Brosh,1 Hila Besserglick,1 Ido Nachmany,3 Yoach Rais,2 Perry Stambolsky,1 Xiaohu Tang,1 Michael Milyavsky,1 Igor Shats,1 Marina Kalis,1 Naomi Goldfinger,1 and Varda Rotter1 1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel; 2Department of Life Science, Bar-Ilan University, Ramat Gan, Israel; and 3Department of General Surgery B, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel Abstract mutated. Notably, the predominant mode of p53 inactivation is by Mutations in p53 are ubiquitous in human tumors. Some p53 point mutation rather than by deletion or truncation. These data mutations not only result in loss of wild-type (WT) activity but coupled with the observation that mutant p53 is generally highly also grant additional functions, termed ‘‘gain of function.’’ overexpressed in tumors have led to the hypothesis that mutant In this study, we explore how the status of p53 affects the p53 possesses gain-of-function activities. This hypothesis is immediate response gene activating transcription factor 3 supported by the results of in vivo and in vitro studies. For (ATF3) in the 12-O-tetradecanoylphorbol-13-acetate (TPA)- example, mice harboring mutant p53 display allele-specific tumor protein kinase C (PKC) pathway. We show that high doses of spectra, higher metastatic frequency, enhanced cell proliferation, TPA induce ATF3 in a WT p53-independent manner correlat- and higher transformation potential compared with their p53-null ingwith PKCs depletion and cell death.
    [Show full text]
  • Action on Muscle Metabolism and Insulin Sensitivity E Strong Enough for a Man, Made for a Woman
    Review The impact of ERa action on muscle metabolism and insulin sensitivity e Strong enough for a man, made for a woman Andrea L. Hevener*, Zhenqi Zhou, Timothy M. Moore, Brian G. Drew, Vicent Ribas ABSTRACT Background: The incidence of chronic disease is elevated in women after menopause. Natural variation in muscle expression of the estrogen receptor (ER)a is inversely associated with plasma insulin and adiposity. Moreover, reduced muscle ERa expression levels are observed in women and animals presenting clinical features of the metabolic syndrome (MetSyn). Considering that metabolic dysfunction impacts nearly a quarter of the U.S. adult population and elevates chronic disease risk including type 2 diabetes, heart disease, and certain cancers, treatment strategies to combat metabolic dysfunction and associated pathologies are desperately needed. Scope of the review: This review will provide evidence supporting a critical and protective role for skeletal muscle ERa in the regulation of metabolic homeostasis and insulin sensitivity, and propose novel ERa targets involved in the maintenance of metabolic health. Major conclusions: Studies identifying ERa-regulated pathways essential for disease prevention will lay the important foundation for the rational design of novel therapeutics to improve the metabolic health of women while limiting secondary complications that have plagued traditional hormone replacement interventions. Ó 2018 Published by Elsevier GmbH. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Keywords Estrogen action; Estrogen receptors; Insulin sensitivity; Metabolic homeostasis 1. INTRODUCTION new-onset T2DM in postmenopausal women following HRT compared with placebo [7]. The mechanism by which HRT reduces T2D incidence For over two decades researchers have shown strong relationships in postmenopausal women is not yet known however molecular between estrogen action and metabolic health in women.
    [Show full text]
  • An Autoregulatory Loop Controls Peroxisome Proliferator-Activated Receptor Γ Coactivator 1Α Expression in Muscle
    Institutional Repository of the University of Basel University Library Schoenbeinstrasse 18-20 CH-4056 Basel, Switzerland http://edoc.unibas.ch/ Year: 2003 An autoregulatory loop controls peroxisome proliferator-activated receptor γ coactivator 1α expression in muscle Handschin, C. and Rhee, J. and Lin, J. and Tarr, P. T. and Spiegelman, B. M. Posted at edoc, University of Basel Official URL: http://edoc.unibas.ch/dok/A5258732 Originally published as: Handschin, C. and Rhee, J. and Lin, J. and Tarr, P. T. and Spiegelman, B. M.. (2003) An autoregulatory loop controls peroxisome proliferator-activated receptor γ coactivator 1α expression in muscle. Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, H. 12. S. 7111-7116. An Autoregulatory Loop Controls PGC-1 Expression in Muscle Christoph Handschin, James Rhee, Jiandie Lin, Paul T. Tarr, and Bruce M. Spiegelman* Dana-Farber Cancer Institute and Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115 Published in Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7111-6. PMID: 12764228. doi: 10.1073/pnas.1232352100 Copyright © National Academy of Sciences; Proceedings of the National Academy of Sciences USA Page 1 of 24 Classification: Biological Sciences, Cell Biology An Autoregulatory Loop Controls PGC-1 Expression in Muscle Christoph Handschin, James Rhee, Jiandie Lin, Paul T. Tarr, and Bruce M. Spiegelman* Dana-Farber Cancer Institute and Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115 * To whom
    [Show full text]
  • Prox1regulates the Subtype-Specific Development of Caudal Ganglionic
    The Journal of Neuroscience, September 16, 2015 • 35(37):12869–12889 • 12869 Development/Plasticity/Repair Prox1 Regulates the Subtype-Specific Development of Caudal Ganglionic Eminence-Derived GABAergic Cortical Interneurons X Goichi Miyoshi,1 Allison Young,1 Timothy Petros,1 Theofanis Karayannis,1 Melissa McKenzie Chang,1 Alfonso Lavado,2 Tomohiko Iwano,3 Miho Nakajima,4 Hiroki Taniguchi,5 Z. Josh Huang,5 XNathaniel Heintz,4 Guillermo Oliver,2 Fumio Matsuzaki,3 Robert P. Machold,1 and Gord Fishell1 1Department of Neuroscience and Physiology, NYU Neuroscience Institute, Smilow Research Center, New York University School of Medicine, New York, New York 10016, 2Department of Genetics & Tumor Cell Biology, St. Jude Children’s Research Hospital, Memphis, Tennessee 38105, 3Laboratory for Cell Asymmetry, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan, 4Laboratory of Molecular Biology, Howard Hughes Medical Institute, GENSAT Project, The Rockefeller University, New York, New York 10065, and 5Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 Neurogliaform (RELNϩ) and bipolar (VIPϩ) GABAergic interneurons of the mammalian cerebral cortex provide critical inhibition locally within the superficial layers. While these subtypes are known to originate from the embryonic caudal ganglionic eminence (CGE), the specific genetic programs that direct their positioning, maturation, and integration into the cortical network have not been eluci- dated. Here, we report that in mice expression of the transcription factor Prox1 is selectively maintained in postmitotic CGE-derived cortical interneuron precursors and that loss of Prox1 impairs the integration of these cells into superficial layers. Moreover, Prox1 differentially regulates the postnatal maturation of each specific subtype originating from the CGE (RELN, Calb2/VIP, and VIP).
    [Show full text]
  • And Represses TNF Gene Expression Activating Transcription Factor-2
    Histone Deacetylase 3, a Class I Histone Deacetylase, Suppresses MAPK11-Mediated Activating Transcription Factor-2 Activation and Represses TNF Gene Expression This information is current as of September 25, 2021. Ulrich Mahlknecht, Jutta Will, Audrey Varin, Dieter Hoelzer and Georges Herbein J Immunol 2004; 173:3979-3990; ; doi: 10.4049/jimmunol.173.6.3979 http://www.jimmunol.org/content/173/6/3979 Downloaded from References This article cites 45 articles, 31 of which you can access for free at: http://www.jimmunol.org/content/173/6/3979.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 25, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2004 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Histone Deacetylase 3, a Class I Histone Deacetylase, Suppresses MAPK11-Mediated Activating Transcription Factor-2 Activation and Represses TNF Gene Expression1 Ulrich Mahlknecht,2* Jutta Will,* Audrey Varin,† Dieter Hoelzer,* and Georges Herbein2† During inflammatory events, the induction of immediate-early genes, such as TNF-␣, is regulated by signaling cascades including the JAK/STAT, NF-␬B, and the p38 MAPK pathways, which result in phosphorylation-dependent activation of transcription factors.
    [Show full text]
  • Targeting Endothelial Kruppel-Like Factor 2 (KLF2) in Arteriovenous
    Targeting Endothelial Krüppel-like Factor 2 (KLF2) in Arteriovenous Fistula Maturation Failure A dissertation submitted to the Graduate School of the University of Cincinnati in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY (Ph.D.) in the Biomedical Engineering Program Department of Biomedical Engineering College of Engineering and Applied Science 2018 by Keith Louis Saum B.S., Wright State University, 2012 Dissertation Committee: Albert Phillip Owens III, Ph.D. (Committee Chair) Begona Campos-Naciff, Ph.D Christy Holland, Ph.D. Daria Narmoneva, Ph.D. Prabir Roy-Chaudhury, M.D., Ph.D Charuhas Thakar, M.D. Abstract The arteriovenous fistula (AVF) is the preferred form of vascular access for hemodialysis. However, 25-60% of AVFs fail to mature to a state suitable for clinical use, resulting in significant morbidity, mortality, and cost for end-stage renal disease (ESRD) patients. AVF maturation failure is recognized to result from changes in local hemodynamics following fistula creation which lead to venous stenosis and thrombosis. In particular, abnormal wall shear stress (WSS) is thought to be a key stimulus which alters endothelial function and promotes AVF failure. In recent years, the transcription factor Krüppel-like factor-2 (KLF2) has emerged as a key regulator of endothelial function, and reduced KLF2 expression has been shown to correlate with disturbed WSS and AVF failure. Given KLF2’s importance in regulating endothelial function, the objective of this dissertation was to investigate how KLF2 expression is regulated by the hemodynamic and uremic stimuli within AVFs and determine if loss of endothelial KLF2 is responsible for impaired endothelial function.
    [Show full text]
  • Global Mef2 Target Gene Analysis in Skeletal and Cardiac Muscle
    GLOBAL MEF2 TARGET GENE ANALYSIS IN SKELETAL AND CARDIAC MUSCLE STEPHANIE ELIZABETH WALES A DISSERTATION SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY GRADUATE PROGRAM IN BIOLOGY YORK UNIVERSITY TORONTO, ONTARIO FEBRUARY 2016 © Stephanie Wales 2016 ABSTRACT A loss of muscle mass or function occurs in many genetic and acquired pathologies such as heart disease, sarcopenia and cachexia which are predominantly found among the rapidly increasing elderly population. Developing effective treatments relies on understanding the genetic networks that control these disease pathways. Transcription factors occupy an essential position as regulators of gene expression. Myocyte enhancer factor 2 (MEF2) is an important transcription factor in striated muscle development in the embryo, skeletal muscle maintenance in the adult and cardiomyocyte survival and hypertrophy in the progression to heart failure. We sought to identify common MEF2 target genes in these two types of striated muscles using chromatin immunoprecipitation and next generation sequencing (ChIP-seq) and transcriptome profiling (RNA-seq). Using a cell culture model of skeletal muscle (C2C12) and primary cardiomyocytes we found 294 common MEF2A binding sites within both cell types. Individually MEF2A was recruited to approximately 2700 and 1600 DNA sequences in skeletal and cardiac muscle, respectively. Two genes were chosen for further study: DUSP6 and Hspb7. DUSP6, an ERK1/2 specific phosphatase, was negatively regulated by MEF2 in a p38MAPK dependent manner in striated muscle. Furthermore siRNA mediated gene silencing showed that MEF2D in particular was responsible for repressing DUSP6 during C2C12 myoblast differentiation. Using a p38 pharmacological inhibitor (SB 203580) we observed that MEF2D must be phosphorylated by p38 to repress DUSP6.
    [Show full text]
  • The Impact of Skeletal Muscle Erα on Mitochondrial Function And
    Copyedited by: oup MINI REVIEW The Impact of Skeletal Muscle ERα on Mitochondrial Function and Metabolic Health Downloaded from https://academic.oup.com/endo/article-abstract/161/2/bqz017/5735479 by University of Southern California user on 19 February 2020 Andrea L. Hevener1,2, Vicent Ribas1, Timothy M. Moore1, and Zhenqi Zhou1 1David Geffen School of Medicine, Department of Medicine, Division of Endocrinology, Diabetes, and Hypertension, University of California, Los Angeles, California 90095; and 2Iris Cantor-UCLA Women’s Health Research Center, University of California, Los Angeles, California 90095 ORCiD numbers: 0000-0003-1508-4377 (A. L. Hevener). The incidence of chronic disease is elevated in women after menopause. Increased expression of ESR1 (the gene that encodes the estrogen receptor alpha, ERα) in muscle is highly associated with metabolic health and insulin sensitivity. Moreover, reduced muscle expression levels of ESR1 are observed in women, men, and animals presenting clinical features of the metabolic syndrome (MetSyn). Considering that metabolic dysfunction elevates chronic disease risk, including type 2 diabetes, heart disease, and certain cancers, treatment strategies to combat metabolic dysfunction and associated pathologies are desperately needed. This review will provide published work supporting a critical and protective role for skeletal muscle ERα in the regulation of mitochondrial function, metabolic homeostasis, and insulin action. We will provide evidence that muscle-selective targeting of ERα may be effective for the preservation of mitochondrial and metabolic health. Collectively published findings support a compelling role for ERα in the control of muscle metabolism via its regulation of mitochondrial function and quality control. Studies identifying ERα-regulated pathways essential for disease prevention will lay the important foundation for the design of novel therapeutics to improve metabolic health of women while limiting secondary complications that have historically plagued traditional hormone replacement interventions.
    [Show full text]
  • The Repression of MEF2 Transcription Factors Exerted by Class Iia Hdacs
    81,9(56,7<2)8',1( BBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBBB 3K'Course in Biomedical Sciences and Biotechnology ;;9,,&<&/( THE REPRESSION OF MEF2 TRANSCRIPTION FACTORS EXERTED BY CLASS IIA HDACS AND THEIR DEGRADATION STIMULATED BY CDK4 DETERMINE THE ACQUISITION OF HALLMARKS OF TRANSFORMATION IN FIBROBLASTS. 3K'6WXGHQW'L*LRUJLR(URV 7XWRUSURI&ODXGLR%UDQFROLQL (URV'L*LRUJLR To my family, Sara and those who believe in the research against cancer ABSTRACT 1 RIASSUNTO 2 INTRODUCTION 3 1. The HDACs world 3 2. Class IIa HDACs: similarities and differences between class IIa and class I HDACs 4 3. Class IIa HDACs: HDACs with orphan substrates or missed during evolution? 7 4. Pathways of regulation 10 a) regulation of class IIa HDACs transcription and modulation of the stability of the messengers (RNAi) 10 b) sub-cellular localization 11 5. Partners and biological functions 20 6. Class IIa HDACs as regulators of proliferation and cancer 28 7. MEF2 family of transcription factors 37 8. On the molecular basis of the MEF2-Class IIa HDACs axis: structure of MEF2/DNA, MEF2/Cabin1/DNA, MEF2/HDAC9/DNA and MEF2/DNA/p300 complexes. 38 9. Pathways of regulation 40 a) binding to repressors and co-activators 41 b) post-translational modifications 42 c) regulation of MEF2s transcription and modulation of the stability of the messengers (RNAi) 49 d) regulation of protein stability 51 10. Main functional roles 53 11. MEF2 as a regulator of differentiation programs 54 12. A lesson from the study of the knock-outs. 56 The role of MEF2s and of the MEF2-class IIa HDACs axis in myogenesis 57 The role of MEF2 TFs in cardiomyogenesis 59 The role of MEF2 TFs and of the MEF2-class IIa HDACs axis in endochondral bone ossification 59 The role of MEF2s and of the MEF2-class IIa HDACs axis in vasculogenesis and differentiation of vascular smooth muscle cells 60 The role of MEF2 TFs in neuronal development 60 The role of MEF2s in hematopoiesis and T cell development 61 The role of MEF2s in melanogenesis 62 The role of MEF2s in neural crest development 62 1.
    [Show full text]
  • The Function of the MEF2 Family of Transcription Factors in Cardiac Development, Cardiogenomics, and Direct Reprogramming
    Journal of Cardiovascular Development and Disease Review The Function of the MEF2 Family of Transcription Factors in Cardiac Development, Cardiogenomics, and Direct Reprogramming Cody A. Desjardins and Francisco J. Naya * Department of Biology, Program in Cell and Molecular Biology, Boston University, 24 Cummington Mall Boston, Boston, MA 02215, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-617-353-2469; Fax: +1-617-353-6340 Academic Editors: Sean M. Wu and Neil C. Chi Received: 5 June 2016; Accepted: 8 August 2016; Published: 11 August 2016 Abstract: Proper formation of the mammalian heart requires precise spatiotemporal transcriptional regulation of gene programs in cardiomyocytes. Sophisticated regulatory networks have evolved to not only integrate the activities of distinct transcription factors to control tissue-specific gene programs but also, in many instances, to incorporate multiple members within these transcription factor families to ensure accuracy and specificity in the system. Unsurprisingly, perturbations in this elaborate transcriptional circuitry can lead to severe cardiac abnormalities. Myocyte enhancer factor–2 (MEF2) transcription factor belongs to the evolutionarily conserved cardiac gene regulatory network. Given its central role in muscle gene regulation and its evolutionary conservation, MEF2 is considered one of only a few core cardiac transcription factors. In addition to its firmly established role as a differentiation factor, MEF2 regulates wide variety of, sometimes antagonistic, cellular processes such as cell survival and death. Vertebrate genomes encode multiple MEF2 family members thereby expanding the transcriptional potential of this core transcription factor in the heart. This review highlights the requirement of the MEF2 family and their orthologs in cardiac development in diverse animal model systems.
    [Show full text]
  • The Nuclear Receptor Pparb/D Programs Muscle Glucose Metabolism in Cooperation with AMPK and MEF2
    Downloaded from genesdev.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press The nuclear receptor PPARb/d programs muscle glucose metabolism in cooperation with AMPK and MEF2 Zhenji Gan,1,3 Eileen M. Burkart-Hartman,2,3,4 Dong-Ho Han,2 Brian Finck,2 Teresa C. Leone,1,2 Emily Y. Smith,1 Julio E. Ayala,1 John Holloszy,2 and Daniel P. Kelly1,2,5 1Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute, Orlando, Florida 32827, USA: 2Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA To identify new gene regulatory pathways controlling skeletal muscle energy metabolism, comparative studies were conducted on muscle-specific transgenic mouse lines expressing the nuclear receptors peroxisome proliferator-activated receptor a (PPARa; muscle creatine kinase [MCK]-PPARa)orPPARb/d (MCK-PPARb/d). MCK-PPARb/d mice are known to have enhanced exercise performance, whereas MCK-PPARa mice perform at low levels. Transcriptional profiling revealed that the lactate dehydrogenase b (Ldhb)/Ldha gene expression ratio is increased in MCK-PPARb/d muscle, an isoenzyme shift that diverts pyruvate into the mitochondrion for the final steps of glucose oxidation. PPARb/d gain- and loss-of-function studies in skeletal myotubes demonstrated that PPARb/d, but not PPARa, interacts with the exercise-inducible kinase AMP-activated protein kinase (AMPK) to synergistically activate Ldhb gene transcription by cooperating with myocyte enhancer factor 2A (MEF2A) in aPPARb/d ligand-independent manner. MCK-PPARb/d muscle was shown to have high glycogen stores, increased levels of GLUT4, and augmented capacity for mitochondrial pyruvate oxidation, suggesting a broad reprogram- ming of glucose utilization pathways.
    [Show full text]
  • The Role of Gata2 in Hematopoietic and Vascular Development By
    The Role of Gata2 in Hematopoietic and Vascular Development by William D Brandt A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Cellular and Molecular Biology) in The University of Michigan 2009 Doctoral Committee: Professor James Douglas Engel, Chair Professor Eric R Fearon Professor Deborah L Gumucio Associate Professor Thomas M Glaser William D Brandt 2009 Dedication To my family, without whom this PhD would never have been possible. ii Acknowledgements The Engel lab and the University of Michigan will always have my deepest gratitude, particularly the lab’s proprietor and my thesis advisor Doug Engel, whose love of science and good nature has always been a source of inspiration. Doug has been instrumental in my growth as a nascent scientist and I will forever be indebted to him. My gratitude also goes to Kim-Chew Lim and Tomo Hosoya, whose wealth of knowledge and support were relied upon regularly. To Deb Gumucio, Tom Glaser, and Eric Fearon, whose advice and support facilitated my maturation from a naïve student to a proficient scientist – thank you. And to Lori Longeway and Kristin Hug, whose capabilities as department representatives I repeatedly put to the test; you came through for me every time. Thank you. Finally, no amount of words can express how truly grateful and indebted I am to my parents and sister – Cary, Kim, and Jenelle. I would not be in this position today without their unerring love and support. iii Table of Contents Dedication ii Acknowledgements iii List of Figures v List of Tables vi Abstract vii Chapter 1.
    [Show full text]