DOCSLIB.ORG

Characterization of the Chimeric Retinoic Acid Receptor RAR /VDR

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Leukemia (1998) 12, 554–562 1998 Stockton Press All rights reserved 0887-6924/98 $12.00 http://www.stockton-press.co.uk/leu Characterization of the chimeric retinoic acid receptor RAR␣/VDR ´ SM Pemrick1, P Abarzua2, C Kratzeisen2, MS Marks3, JA Medin3, K Ozato3 and JF Grippo1 Departments of 1Metabolic Diseases and 2Oncology, Hoffmann-La Roche, Inc., Nutley, NJ; and 3Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA The chimeric receptor, RAR␣/VDR, contains the DNA-binding superfamily are dimeric, ligand-inducible-transcription fac- ␣ domain of the retinoic acid receptor (RAR ) and the ligand- tors, which activate nuclear target genes by binding to specific binding domain of the vitamin D receptor (VDR). The ligand- binding properties of RAR␣/VDR are equivalent to that of VDR, DNA sequences, termed HREs, within the promoter region. ␣ with an observed Kd for 1 ,25 dihydroxy-vitamin D3 (D3)of HREs are arranged as palindromic or direct repeats (DR) of a 0.5 nM. In CV-1 cells, both RAR␣ and RAR␣/VDR induce compa- hexad consensus sequence (PuGGTCA). The steroid receptors, rable levels of ligand-mediated transcriptional activity from the such as GR and ER, bind as homodimers to palindromic HREs, ␤ retinoic acid responsive reporter gene, (RARE)3-TK-lucifer- whereas RAR, TR and VDR (the non-steroid receptors) form ase, in the presence of the ligand predicted from the receptor ligand-binding domain. Two chimeric RAR receptors were con- heterodimers with RXR and bind preferentially to DR HREs, structed which contained the ligand-binding domain of the each heterodimer selecting for a specific distance between estrogen receptor (ER): RAR␣/ER and ER/RAR␣/ER. Both hexad repeats (for review see Refs 11 and 12). RAR␣/ER and ER/RAR␣/ER bind ␤-estradiol with high affinity, To fine tune retinoid-induced gene regulation, there are and are transcriptionally active only from palindromic RAREs ␣ RAR and RXR subtypes and numerous isoforms, which are (TREpal and/or (TRE3)3). Only RAR /VDR matched in kind and 11 degree the functional characteristics of RAR␣: (1) maximally conserved more across species than within a species. One active from the ␤(RARE); (2) moderately active from the TREs; hypothesis states dimeric combinations of RAR and RXR sub- (3) inactive from the retinoic X receptor response elements types and isoforms regulate subsets of RAREs to exert the plei- (RXREs) ApoA1 and CRBP II; (4) forms heterodimers with otropic effects of retinoids.13 Approaches to explore this ␣ ␤ RXR ; and (5) binds to the RARE. F9 embryonal carcinoma hypothesis have included: rational development of receptor- cell lines were generated which express RAR␣/VDR mRNA (F9- RAR␣/VDR cells) and compared with F9 wild-type (F9-Wt) cells, specific ligands (eg see Ref. 14); targeted disruption which do not express VDR mRNA. Treatment with all-trans reti- (‘knockout’) via homologous recombination of one or both noic acid (tRA) inhibits cell growth and induces the differen- alleles for a particular receptor subtype.2,15 In addition, recep- tiation morphology in both F9-Wt and F9-RAR␣/VDR cells; tor-based gene therapies could feasibly be developed to whereas, treatment with D3 is similarly effective only for F9- manipulate the threshold concentration of specific RXR-RAR RAR␣/VDR cells. It is concluded RAR␣/VDR is an useful ‘tool’ to pinpoint, or to augment transcription from RAREs in gene heterodimers in cancers responsive to retinoids. For the chim- pathways controlled by RAR without inhibiting the retinoid era, RAR␣/VDR described here, the inducible ligand would responsiveness of endogenous RARs. be D3, eliminating acquired drug resistance due to the pharm- Keywords: retinoic acid receptors (RARs) and chimeras; all-trans acokinetic properties of retinoids. retinoic acid; vitamin D3; permanent gene transfection and Designing functional chimeric receptors can be a straight- expression; terminal differentiation; leukemia forward experiment because all hormone receptors possess a linearly arranged modular structure (regions A to E or F), con- served to varying degrees within each family.16 For example, Introduction the chimera GR/TR, replaced the n-terminal A/B region of TR for that of GR, to enhance T induction of transcriptional acti- All-trans retinoic acid (tRA) is known to be a key regulator of 3 vation.17 The most conserved region, the C or DNA-binding growth and development in both the adult, from vitamin A domain, has two zinc finger motifs, and recognizes the base deficient (VAD) animal studies, and in the embryo, from 18 analysis of the teratogenic effects resulting from administration pair sequence of the HRE. In a now classical experiment, 1,2 the glucocorticoid HRE became responsive to ␤-estradiol by of exogenous tRA. Similarly, tRA induces differentiation and 19 inhibits proliferation of certain cell types, including HL-60 replacing the C region of ER with that of GR. Region E, the leukemia3 and F9 embryonal carcinoma cells.4,5 tRA is being ligand-binding domain, is also involved in other structurally 20–24 22 explored as a differentiation therapy in cancer because it overlapping functions, including transactivation and 23,24 induces complete remission in acute promyelocytic leukemia dimerization. The receptor polarity of DR HREs, with RXR ′ (APL).6 Unfortunately, APL patients receiving only tRA therapy at the 5 HRE half-site, plus the freedom of rotation about the ′ 25 eventually experience retinoid resistance and clinical hinge region of the 3 -dimeric partner, suggests the ligand- relapse.7,8 binding domains of RAR, VDR, and TR may be functionally At the molecular level, tRA is a ligand for the retinoic acid interchangeable. Indeed, the ligand-binding domain of TR can receptor, RAR, which also binds the RA isomer 9-cis-RA substitute for that of RAR to place both synthetic25 and natural 9,10 26 (9cRA). Related receptors, RXRs, bind 9cRA. The RARs and RAREs under the control of T3. We describe in this report, RXRs belong to the steroid/nuclear receptor superfamily, the functional characteristics of the chimeric receptor which also includes the thyroid (TR), vitamin D (VDR), gluco- RAR␣/VDR, and show in F9 cells that RAR␣/VDR can mediate corticoid (GR), and estrogen (ER) receptors. Members of this the differentiation morphology in response to D3 treatment, without disturbing retinoid responsiveness. We conclude RAR␣/VDR can be a ‘tool’ to pinpoint or to modulate gene Correspondence: SM Pemrick at her present address: Merck Research pathways regulated by RARs, without inhibiting the functional Laboratories, RY32-605, Rahway, NJ 07065-0900, USA; Fax: penetrance of endogenous RARs. 518 392 6665 Received 25 April 1997; accepted 21 November 1997 Functional properties of RAR␣/VDR SM Pemrick et al 555 Materials and methods (225 cm2 flasks) and screened for VDR and RAR␣/VDR mRNA. Materials tRA and vitamin D3 were obtained from the Department of Growth and differentiation of F9 cells Medicinal Chemistry, Hoffmann-La Roche; G418 sulfate from Gibco BRL (Grand Island, NY, USA); dibutyryl cAMP and F9-Wt and F9-RAR␣/VDR cell lines were plated (1–2 × 104 theophylline from Sigma Chemical (St Louis, MO, USA); cells/well) in 1 ml culture medium in gelatinized 24-well ␣ 3 1 ,25-[26,27- H]-dihydroxy vitamin D3 (D3) specific activity Costar dishes (Cambridge, MA, USA). After 24 h, 10– 35 (sp. ac.) 155 Ci/mmol (NET-626) and S-methionine were 100 ␮l/well of filtered drug stock solution in phosphate-buff- purchased from Dupont-NEN (Boston, MA, USA); [2,4,6,7- ered saline (PBS)-Ca2+/Mg2+ free was added to a final concen- 3H]-oestradiol, sp. ac. 92 Ci/mmol (TRE.322), and deoxycytid- ␮ ␮ tration of 0.1% ethanol, 1 M tRA, 30 or 100 nM D3, 250 M ′ ␣32 ine-5 P triphosphate, sp. ac. 3000 Ci/mmol (AA005) from dibutyryl cAMP, 500 ␮M theophylline. The cultures were Amersham Life Sciences (Arlington Heights, IL, USA). HPLC incubated for 96 h, washed with PBS, photographed (through analysis placed ligand purity at greater than 97%. an inverted microscope equipped with phase contrast optics), trypsinized, and counted manually by means of a hemacyto- meter. Plasmid construction Annealed oligo nucleotides for the following HREs were RNA preparation and Northern analysis inserted into either the BglII or the BamHI site of pTK-lucifer- 27 ␤ ase (LUC); (RARE)3, gatc(gggtagGGTTCAccgaaAGTTCA Total RNA was extracted from F9 cells by the guanidine iso- ctcg)3; (TRE3)3, gatc(ttAGGTCAgggacgTGACCTaa)3aaggccta; thiocyanate procedure (RNAzol B; Biotecx Laboratories, Hou- 11 ′ TREpal, gatctcAGGTCATGACCTga; ERE, the 5 -flanking ston, TX, USA). For Northern analysis, total RNA (20–30 ␮g) − − 28 region ( 331 to 297) of the Xenopus vitellogenin A2 gene. from F9 clones was size fractionated on 1% agarose/6% for- For RXRE(CRBPII)-SV-LUC, the RXRE (gatctgCTGTCAc maldehyde gels in 20 mM MOPS (pH 7.0 HCl), 50 mM Na AGGTCAc AGGTCAcAGGTCAcAGTT),11 was inserted into 2 + EDTA, 5 mM sodium acetate and transferred to Gene Screen the BglII cloning site of Blue Script IIKS( ) (Stratagene, La Jolla, nylon membranes (DuPont-NEN). The RNA blots were CA, USA) in front of the LUC gene and SV40 promoter. The hybridized (at 42°C) with a random primer labeled cDNA RXRE(ApoA1)-LUC reporter gene had a 490 base pair HindIII probe (Boehringer Mannheim Random Primed DNA Labeling fragment of [1Xa]-41A1.CAT containing the apoA1 enhancer 29 Kit; Indianapolis, IN, USA), for the ligand-binding domain of site A oligo A inserted in the HindIII cloning site of the VDR (Figure 2a), which had been purified on G50 sephadex pGL2-basic vector (Promega, Madison, WI, USA). The oligo Quick Spin Columns (Boehringer Mannheim). Washed blots A sequence was: gatcaTGACCCctTGAACCc TGTCCTgatc. For ␤ were exposed to Kodak X-Omat film with an intensifying p Ac-lacZ (gift of G Vasios, Osteoarthritis Sciences Inc, Cam- screen at −80°C, for varying amounts of time.
Recommended publications
  • Retinoic Acid Signaling and Neuronal Differentiation

    Retinoic Acid Signaling and Neuronal Differentiation

    Cell. Mol. Life Sci. (2015) 72:1559–1576 DOI 10.1007/s00018-014-1815-9 Cellular and Molecular Life Sciences REVIEW Retinoic acid signaling and neuronal differentiation Amanda Janesick • Stephanie Cherie Wu • Bruce Blumberg Received: 23 October 2014 / Revised: 15 December 2014 / Accepted: 19 December 2014 / Published online: 6 January 2015 Ó Springer Basel 2015 Abstract The identification of neurological symptoms cell cycle exit downstream of RA will be critical for our caused by vitamin A deficiency pointed to a critical, early understanding of how to target tumor differentiation. developmental role of vitamin A and its metabolite, reti- Overall, elucidating the molecular details of RAR-regu- noic acid (RA). The ability of RA to induce post-mitotic, lated neurogenesis will be decisive for developing and neural phenotypes in various stem cells, in vitro, served as understanding neural proliferation–differentiation switches early evidence that RA is involved in the switch between throughout development. proliferation and differentiation. In vivo studies have expanded this ‘‘opposing signal’’ model, and the number of Keywords Neurogenesis Á Retinoic acid receptor Á primary neurons an embryo develops is now known to Proliferation-differentiation switch depend critically on the levels and spatial distribution of RA. The proneural and neurogenic transcription factors that control the exit of neural progenitors from the cell Introduction cycle and allow primary neurons to develop are partly elucidated, but the downstream effectors of RA receptor The role of retinoic acid (RA) in neurogenesis has been (RAR) signaling (many of which are putative cell cycle known indirectly for as long as haliver (halibut) and cod liver regulators) remain largely unidentified.
  • Homeobox Gene Expression Profile in Human Hematopoietic Multipotent

    Homeobox Gene Expression Profile in Human Hematopoietic Multipotent

    Leukemia (2003) 17, 1157–1163 & 2003 Nature Publishing Group All rights reserved 0887-6924/03 $25.00 www.nature.com/leu Homeobox gene expression profile in human hematopoietic multipotent stem cells and T-cell progenitors: implications for human T-cell development T Taghon1, K Thys1, M De Smedt1, F Weerkamp2, FJT Staal2, J Plum1 and G Leclercq1 1Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium; and 2Department of Immunology, Erasmus Medical Center, Rotterdam, The Netherlands Class I homeobox (HOX) genes comprise a large family of implicated in this transformation proces.14 The HOX-C locus transcription factors that have been implicated in normal and has been primarily implicated in lymphomas.15 malignant hematopoiesis. However, data on their expression or function during T-cell development is limited. Using degener- Hematopoietic cells are derived from stem cells that reside in ated RT-PCR and Affymetrix microarray analysis, we analyzed fetal liver (FL) in the embryo and in the adult bone marrow the expression pattern of this gene family in human multipotent (ABM), which have the unique ability to self-renew and thereby stem cells from fetal liver (FL) and adult bone marrow (ABM), provide a life-long supply of blood cells. T lymphocytes are a and in T-cell progenitors from child thymus. We show that FL specific type of hematopoietic cells that play a major role in the and ABM stem cells are similar in terms of HOX gene immune system. They develop through a well-defined order of expression, but significant differences were observed between differentiation steps in the thymus.16 Several transcription these two cell types and child thymocytes.
  • Signaling by Retinoic Acid in Embryonic and Adult Hematopoiesis

    Signaling by Retinoic Acid in Embryonic and Adult Hematopoiesis

    J. Dev. Biol. 2014, 2, 18-33; doi:10.3390/jdb2010018 OPEN ACCESS Journal of Developmental Biology ISSN 2221-3759 www.mdpi.com/journal/jdb/ Review Signaling by Retinoic Acid in Embryonic and Adult Hematopoiesis Elena Cano †, Laura Ariza †, Ramón Muñoz-Chápuli and Rita Carmona * Department of Animal Biology, Faculty of Science, University of Málaga, E29071 Málaga, Spain; E-Mails: [email protected] (E.C.); [email protected] (L.A.); [email protected] (R.M.-C.) † These authors contributed equally to this paper. * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +34-952-134-135; Fax: +34-952-131-668. Received: 3 February 2014; in revised form: 5 March 2014 / Accepted: 6 March 2014 / Published: 17 March 2014 Abstract: Embryonic and adult hematopoiesis are both finely regulated by a number of signaling mechanisms. In the mammalian embryo, short-term and long-term hematopoietic stem cells (HSC) arise from a subset of endothelial cells which constitute the hemogenic endothelium. These HSC expand and give rise to all the lineages of blood cells in the fetal liver, first, and in the bone marrow from the end of the gestation and throughout the adult life. The retinoic acid (RA) signaling system, acting through the family of nuclear retinoic acid receptors (RARs and RXRs), is involved in multiple steps of the hematopoietic development, and also in the regulation of the differentiation of some myeloid lineages in adults. In humans, the importance of this RA-mediated control is dramatically illustrated by the pathogeny of acute promyelocytic leukemia, a disease produced by a chromosomal rearrangement fusing the RAR gene with other genes.
  • Pathway Development Via Retinoid X Receptor Vitamin a Enhances In

    Pathway Development Via Retinoid X Receptor Vitamin a Enhances In

    Vitamin A Enhances in Vitro Th2 Development Via Retinoid X Receptor Pathway This information is current as Charles B. Stephensen, Reuven Rasooly, Xiaowen Jiang, of September 24, 2021. Michael A. Ceddia, Casey T. Weaver, Roshantha A. S. Chandraratna and R. Patterson Bucy J Immunol 2002; 168:4495-4503; ; doi: 10.4049/jimmunol.168.9.4495 http://www.jimmunol.org/content/168/9/4495 Downloaded from References This article cites 40 articles, 24 of which you can access for free at: http://www.jimmunol.org/content/168/9/4495.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 24, 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 © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Vitamin A Enhances in Vitro Th2 Development Via Retinoid X Receptor Pathway1 Charles B. Stephensen,2* Reuven Rasooly,* Xiaowen Jiang,* Michael A. Ceddia,3* Casey T. Weaver,† Roshantha A. S. Chandraratna,‡ and R. Patterson Bucy† Vitamin A deficiency diminishes Th2-mediated Ab responses, and high-level dietary vitamin A or treatment with the vitamin A metabolite retinoic acid (RA) enhances such responses.
  • Isolation and Characterisation of Lymphatic Endothelial Cells From

    Isolation and Characterisation of Lymphatic Endothelial Cells From

    www.nature.com/scientificreports OPEN Isolation and characterisation of lymphatic endothelial cells from lung tissues afected by lymphangioleiomyomatosis Koichi Nishino1,2*, Yasuhiro Yoshimatsu3,4, Tomoki Muramatsu5, Yasuhito Sekimoto1,2, Keiko Mitani1,2, Etsuko Kobayashi1,2, Shouichi Okamoto1,2, Hiroki Ebana1,2,6,7, Yoshinori Okada8, Masatoshi Kurihara2,6, Kenji Suzuki9, Johji Inazawa5, Kazuhisa Takahashi1, Tetsuro Watabe3 & Kuniaki Seyama1,2 Lymphangioleiomyomatosis (LAM) is a rare pulmonary disease characterised by the proliferation of smooth muscle-like cells (LAM cells), and an abundance of lymphatic vessels in LAM lesions. Studies reported that vascular endothelial growth factor-D (VEGF-D) secreted by LAM cells contributes to LAM-associated lymphangiogenesis, however, the precise mechanisms of lymphangiogenesis and characteristics of lymphatic endothelial cells (LECs) in LAM lesions have not yet been elucidated. In this study, human primary-cultured LECs were obtained both from LAM-afected lung tissues (LAM-LECs) and normal lung tissues (control LECs) using fuorescence-activated cell sorting (FACS). We found that LAM-LECs had signifcantly higher ability of proliferation and migration compared to control LECs. VEGF-D signifcantly promoted migration of LECs but not proliferation of LECs in vitro. cDNA microarray and FACS analysis revealed the expression of vascular endothelial growth factor receptor (VEGFR)-3 and integrin α9 were elevated in LAM-LECs. Inhibition of VEGFR-3 suppressed proliferation and migration of LECs, and blockade of integrin α9 reduced VEGF-D-induced migration of LECs. Our data uncovered the distinct features of LAM-associated LECs, increased proliferation and migration, which may be due to higher expression of VEGFR-3 and integrin α9. Furthermore, we also found VEGF-D/VEGFR-3 and VEGF-D/ integrin α9 signaling play an important role in LAM-associated lymphangiogenesis.
  • Peroxisome Proliferator-Activated Receptor ; Is Highly Expressed In

    Peroxisome Proliferator-Activated Receptor ; Is Highly Expressed In

    Imaging, Diagnosis, Prognosis Peroxisome Proliferator-Activated Receptor ; Is Highly Expressed in Pancreatic Cancer and Is Associated With Shorter Overall Survival Times Glen Kristiansen,1Juliane Jacob,1Ann-Christin Buckendahl,1Robert Gru« tzmann,3 Ingo Alldinger,3 Bence Sipos,4 Gu« nter Klo« ppel,4 Marcus Bahra,2 Jan M. Langrehr,2 Peter Neuhaus,2 Manfred Dietel,1and Christian Pilarsky3 Abstract Purpose: Peroxisome proliferator-activated receptor g (PPARg) is a ligand-activated transcrip- tion factor that has been implicated in carcinogenesis and progression of various solid tumors, including pancreatic carcinoma.We aimed to clarify the expression patterns of PPARg in pancre- atic ductal carcinomas and to correlate these to clinicopathologic variables, including patient survival. Experimental Design: Array-based expression profiling of 19 microdissected carcinomas and 14 normal ductal epithelia was conducted. Additionally,Western blots of pancreatic cancer cell lines and paraffinized tissue of 129 pancreatic carcinomas were immunostained for PPARg.For statistical analysis, Fisher’s exact test, m2 test for trends, correlation analysis, Kaplan-Meier analysis, and Cox’s regression were applied. Results: Expression profiles showed a strong overexpression of PPARg mRNA (change fold, 6.9; P = 0.04). Immunohistochemically, PPARg expression was seen in 71.3% of pancreatic cancer cases. PPARg expression correlated positively to higher pTstages and higher tumor grade. Survival analysis showed a significant prognostic value for PPARg, which was found to be independent in the clinically important subgroup of node-negative tumors. Conclusions: PPARg is commonly up-regulated in pancreatic ductal adenocarcinoma and might be a prognostic marker in this disease. Both findings corroborate the importance of PPARg in tumor progression of pancreatic cancer.
  • The First Genome-Wide View of Vitamin D Receptor Locations and Their Mechanistic Implications

    The First Genome-Wide View of Vitamin D Receptor Locations and Their Mechanistic Implications

    ANTICANCER RESEARCH 32: 271-282 (2012) Review The First Genome-wide View of Vitamin D Receptor Locations and Their Mechanistic Implications CARSTEN CARLBERG1, SABINE SEUTER2 and SAMI HEIKKINEN1 1Department of Biosciences, University of Eastern Finland, Kuopio, Finland; 2Life Sciences Research Unit, University of Luxembourg, Luxembourg, Luxembourg Abstract. The transcription factor vitamin D receptor Physiological Impact of Vitamin D (VDR) is the nuclear sensor for the biologically most active in the Immune System metabolite of vitamin D, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3). The physiological actions of the VDR and Vitamin D is a micronutrient which under ultraviolet (UV) its ligand are not only the well-known regulation of calcium radiation can also be produced in the skin (1). The most and phosphorus uptake and transport controlling bone abundant form of vitamin D is its liver hydroxylation product formation, but also their significant involvement in the 25-hydroxyvitamin D3 (25(OH)D3), serum concentrations of control of immune functions and of cellular growth and which indicate the vitamin D status of the human individual differentiation. For a general understanding of the (2). The biologically most active vitamin D metabolite is mechanisms of 1α,25(OH)2D3 signaling, it is essential to obtained from further hydroxylation of 25(OH)D3 in the monitor the genome-wide location of VDR in relation to kidney to 1α,25(OH)2D3 (3). Interestingly, the hydroxylation primary 1α,25(OH)2D3 target genes. Within the last months, of vitamin D can also take place in other tissues and a few of two chromatin immunoprecipitation sequencing (ChIP-Seq) them, such as keratinocytes and macrophages, have the studies using cells of the hematopoietic system, capacity for the full conversion of vitamin D to lymphoblastoids and monocytes, were published.
  • Drosophila Pax6 Promotes Development of the Entire Eye-Antennal Disc, Thereby Ensuring Proper Adult Head Formation

    Drosophila Pax6 Promotes Development of the Entire Eye-Antennal Disc, Thereby Ensuring Proper Adult Head Formation

    PAPER Drosophila Pax6 promotes development of the entire COLLOQUIUM eye-antennal disc, thereby ensuring proper adult head formation Jinjin Zhua, Sneha Palliyila, Chen Ranb, and Justin P. Kumara,1 aDepartment of Biology, Indiana University, Bloomington, IN 47405; and bDepartment of Biology, Stanford University, Stanford, CA 94305 Edited by Ellen V. Rothenberg, California Institute of Technology, Pasadena, CA, and accepted by Editorial Board Member Neil H. Shubin February 17, 2017 (received for review July 26, 2016) Paired box 6 (Pax6) is considered to be the master control gene for molecular battle among GRNs allows for the subdivision of the eye development in all seeing animals studied so far. In vertebrates, eye-antennal disc to be maintained within a single continuous it is required not only for lens/retina formation but also for the cellular field (13–16). Of the GRNs that are known to operate development of the CNS, olfactory system, and pancreas. Although within the eye-antennal disc, the retinal determination (RD) Pax6 plays important roles in cell differentiation, proliferation, and network, which controls eye development, is the best studied (17). patterning during the development of these systems, the underlying At the core of the RD network lie the Paired box 6 (Pax6) genes mechanism remains poorly understood. In the fruit fly, Drosophila eyeless (ey)andtwin of eyeless (toy), the SIX family member sine melanogaster, Pax6 also functions in a range of tissues, including oculis (so), the transcriptional coactivator eyes absent (eya), and the the eye and brain. In this report, we describe the function of Pax6 in Ski/Sno family member dachshund (dac)(17).
  • The Histone Acetylase PCAF Is a Nuclear Receptor Coactivator

    The Histone Acetylase PCAF Is a Nuclear Receptor Coactivator

    Downloaded from genesdev.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press The histone acetylase PCAF is a nuclear receptor coactivator Jorge C.G. Blanco,1,4 Saverio Minucci,1 Jianming Lu,1 Xiang-Jiao Yang,1 Kristen K. Walker,3 Hongwu Chen,3 Ronald M. Evans,2,3 Yoshihiro Nakatani,1 and Keiko Ozato1,5 1Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, National Institutes of Health (NIH), Bethesda, Maryland 20892-2753 USA; 2Howard Hughes Medical Institute; 3The Salk Institute for Biological Studies, La Jolla, California 92037 USA Whereas the histone acetylase PCAF has been suggested to be part of a coactivator complex mediating transcriptional activation by the nuclear hormone receptors, the physical and functional interactions between nuclear receptors and PCAF have remained unclear. Our efforts to clarify these relationships have revealed two novel properties of nuclear receptors. First, we demonstrate that the RXR/RAR heterodimer directly recruits PCAF from mammalian cell extracts in a ligand-dependent manner and that increased expression of PCAF leads to enhanced retinoid-responsive transcription. Second, we demonstrate that, in vitro, PCAF directly associates with the DNA-binding domain of nuclear receptors, independently of p300/CBP binding, therefore defining a novel cofactor interaction surface. Furthermore, our results show that dissociation of corepressors enables ligand-dependent PCAF binding to the receptors. This observation illuminates how a ligand-dependent receptor function can be propagated to regions outside the ligand-binding domain itself. On the basis of these observations, we suggest that PCAF may play a more central role in nuclear receptor function than previously anticipated.
  • LSD1 Dual Function in Mediating Epigenetic Corruption of the Vitamin

    LSD1 Dual Function in Mediating Epigenetic Corruption of the Vitamin

    Battaglia et al. Clinical Epigenetics (2017) 9:82 DOI 10.1186/s13148-017-0382-y RESEARCH Open Access LSD1 dual function in mediating epigenetic corruption of the vitamin D signaling in prostate cancer Sebastiano Battaglia1*, Ellen Karasik2, Bryan Gillard2, Jennifer Williams2, Trisha Winchester3, Michael T. Moser2, Dominic J Smiraglia3 and Barbara A. Foster2* Abstract Background: Lysine-specific demethylase 1A (LSD1) is a key regulator of the androgen (AR) and estrogen receptors (ER), and LSD1 levels correlate with tumor aggressiveness. Here, we demonstrate that LSD1 regulates vitamin D receptor (VDR) activity and is a mediator of 1,25(OH)2-D3 (vitamin D) action in prostate cancer (PCa). Methods: Athymic nude mice were xenografted with CWR22 cells and monitored weekly after testosterone pellet removal. Expression of LSD1 and VDR (IHC) were correlated with tumor growth using log-rank test. TRAMP tumors and prostates from wild-type (WT) mice were used to evaluate VDR and LSD1 expression via IHC and western blotting. The presence of VDR and LSD1 in the same transcriptional complex was evaluated via immunoprecipitation (IP) using nuclear cell lysate. The effect of LSD1 and 1,25(OH)2-D3 on cell viability was evaluated in C4-2 and BC1A cells via trypanblueexclusion.TheroleofLSD1inVDR-mediatedgenetranscriptionwasevaluatedforCdkn1a, E2f1, Cyp24a1,andS100g via qRT-PCR-TaqMan and via chromatin immunoprecipitation assay. Methylation of Cdkn1a TSS was measured via bisulfite sequencing, and methylation of a panel of cancer-related genes was quantified using methyl arrays. The Cancer Genome Atlas data were retrieved to identify genes whose status correlates with LSD1 and DNA methyltransferase 1 (DNMT1).
  • The Receptor Interacting Protein 1 Inhibits P53 Induction Through NF-KB Activation and Confers a Worse Prognosis in Glioblastoma

    The Receptor Interacting Protein 1 Inhibits P53 Induction Through NF-KB Activation and Confers a Worse Prognosis in Glioblastoma

    Research Article The Receptor Interacting Protein 1 Inhibits p53 Induction through NF-KB Activation and Confers a Worse Prognosis in Glioblastoma Seongmi Park,1 Kimmo J. Hatanpaa,2,7 Yang Xie,3,8 Bruce E. Mickey,4,7 Christopher J. Madden,4,7 Jack M. Raisanen,2,7 Deepti B. Ramnarain,1 Guanghua Xiao,3 Debabrata Saha,5 David A. Boothman,8 Dawen Zhao,6 Robert M. Bachoo,1,7,8 Russell O. Pieper,9 and Amyn A. Habib1,7,8 Departments of 1Neurology, 2Pathology, 3Clinical Sciences, 4Neurosurgery, 5Radiation Oncology, and 6Radiology, 7Annette G. Strauss Center of Neurooncology, and 8Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas and 9Department of Neurological Surgery, University of California-San Francisco, San Francisco, California Abstract studies have linked components of the NF-nB signaling pathway to Nuclear factor-KB (NF-KB) activation may play an important cell cycle progression and tumorigenesis (11–16). role in the pathogenesis of cancer and also in resistance to An intriguing mechanism underlying the pathogenesis of treatment. Inactivation of the p53 tumor suppressor is a key inflammation-induced cancer is the negative regulation of tumor component of the multistep evolution of most cancers. Links suppressor pathways by inflammatory and stress-induced signals. between the NF-KB and p53 pathways are under intense p53 is a key tumor suppressor altered in a broad range of human investigation. In this study, we show that the receptor cancers, including glioma, and an important outcome of p53 interacting protein 1 (RIP1), a central component of the activation is cell cycle arrest or apoptosis after DNA damage K (17, 18).
  • Retinoic Acid-Related Orphan Receptor Rorβ, Circadian Rhythm Abnormalities and Tumorigenesis (Review)

    Retinoic Acid-Related Orphan Receptor Rorβ, Circadian Rhythm Abnormalities and Tumorigenesis (Review)

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 35: 1493-1500, 2015 Retinoic acid-related orphan receptor RORβ, circadian rhythm abnormalities and tumorigenesis (Review) SHUJIONG FENG1, SONG XU1, ZHENZHEN WEN1 and YONGLIANG ZHU1,2 1Laboratory of Gastroenterology, The Second Affiliated Hospital of Zhejiang University, School of Medicine; 2Cancer Institute and Education Ministry Key Laboratory of Cancer Prevention and Intervention, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China Received August 8, 2014; Accepted March 12, 2015 DOI: 10.3892/ijmm.2015.2155 Abstract. Nuclear receptors are a superfamily of transcription and have important physiological functions in cell develop- factors including the steroid hormone receptors, non-steroid ment and differentiation, circadian rhythm, metabolism and hormone receptors and the orphan nuclear receptor family. immune regulation. NRs consist of three components: the Retinoic acid-related orphan receptor (ROR)β, as a member of steroid hormone receptors, non-steroid hormone receptors the orphan nuclear receptor family, plays an important regula- and the orphan nuclear receptor family. Steroid and non- tory role in the maintenance of a variety of physiological and steroid hormone receptors have specific ligands, including pathological processes. RORβ has been determined to act as steroid hormones, thyroid hormones, retinoic acids and fatty an osteogenic repressor in regulating bone formation, and is acids. Ligands for orphan NRs have not yet been determined. involved in regulating circadian rhythm. The findings of recent Retinoic acid-related orphan receptors (RORs), also known studies concerning the association between tumorigenesis as nuclear receptor subfamily 1 group F members (NR1F), and circadian rhythm have shown that an aberrant circadian are specified by gene sequences, which are homologous to rhythm may promote tumorigenesis and tumor progression.