Probing Dominant Negative Behavior of Glucocorticoid Receptor β through a Hybrid Structural and Biochemical Approach
Abstract
Glucocorticoid receptor β (GRβ) is associated with glucocorticoid resistance via dominant negative regulation of GRα. To better understand how GRβ functions as a dominant negative inhibitor of GRα at a molecular level, we determined the crystal structure of the ligand binding domain of GRβ complexed with the antagonist RU-486. The structure reveals that GRβ binds RU-486 in the same ligand binding pocket as GRα, and furthermore, the unique C-terminal amino acids of GRβ are mostly disordered. Binding energy analysis suggests that these C-terminal residues of GRβ do not contribute to RU-486 binding. Intriguingly, the GRβ/RU-486 complex binds corepressor peptide with affinity similar to that of a GRα/RU-486 complex, despite the lack of helix 12. Our biophysical and biochemical analyses reveal that in the presence of RU-486, GRβ is found in a conformation that favors corepressor binding, potentially antagonizing GRα function. This study thus presents an unexpected molecular mechanism by which GRβ could repress transcription.
- Authors:
-
[1]
- National Inst. of Health (NIH), Research Triangle Park, NC (United States). National Inst. of Environmental Health Sciences, Genome Integrity and Structural Biology Lab.
- National Inst. of Health (NIH), Research Triangle Park, NC (United States). National Inst. of Environmental Health Sciences, Signal Transduction Lab.
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- National Institutes of Health (NIH), National Institute of Environmental Health Sciences; USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1435832
- Grant/Contract Number:
- ZIA ES102645; Z01 ES043010; Z01 ES090057; W-31-109-Eng-38
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Molecular and Cellular Biology
- Additional Journal Information:
- Journal Volume: 38; Journal Issue: 8; Journal ID: ISSN 0270-7306
- Publisher:
- American Society for Microbiology
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; glucocorticoid receptor beta (GRβ), RU-486; dominant negative; glucocorticoid resistance; transrepression
Citation Formats
Min, Jungki, Perera, Lalith, Krahn, Juno M., Jewell, Christine M., Moon, Andrea F., Cidlowski, John A., and Pedersen, Lars C. Probing Dominant Negative Behavior of Glucocorticoid Receptor β through a Hybrid Structural and Biochemical Approach. United States: N. p., 2018.
Web. doi:10.1128/MCB.00453-17.
Min, Jungki, Perera, Lalith, Krahn, Juno M., Jewell, Christine M., Moon, Andrea F., Cidlowski, John A., & Pedersen, Lars C. Probing Dominant Negative Behavior of Glucocorticoid Receptor β through a Hybrid Structural and Biochemical Approach. United States. https://doi.org/10.1128/MCB.00453-17
Min, Jungki, Perera, Lalith, Krahn, Juno M., Jewell, Christine M., Moon, Andrea F., Cidlowski, John A., and Pedersen, Lars C. Mon .
"Probing Dominant Negative Behavior of Glucocorticoid Receptor β through a Hybrid Structural and Biochemical Approach". United States. https://doi.org/10.1128/MCB.00453-17. https://www.osti.gov/servlets/purl/1435832.
@article{osti_1435832,
title = {Probing Dominant Negative Behavior of Glucocorticoid Receptor β through a Hybrid Structural and Biochemical Approach},
author = {Min, Jungki and Perera, Lalith and Krahn, Juno M. and Jewell, Christine M. and Moon, Andrea F. and Cidlowski, John A. and Pedersen, Lars C.},
abstractNote = {Glucocorticoid receptor β (GRβ) is associated with glucocorticoid resistance via dominant negative regulation of GRα. To better understand how GRβ functions as a dominant negative inhibitor of GRα at a molecular level, we determined the crystal structure of the ligand binding domain of GRβ complexed with the antagonist RU-486. The structure reveals that GRβ binds RU-486 in the same ligand binding pocket as GRα, and furthermore, the unique C-terminal amino acids of GRβ are mostly disordered. Binding energy analysis suggests that these C-terminal residues of GRβ do not contribute to RU-486 binding. Intriguingly, the GRβ/RU-486 complex binds corepressor peptide with affinity similar to that of a GRα/RU-486 complex, despite the lack of helix 12. Our biophysical and biochemical analyses reveal that in the presence of RU-486, GRβ is found in a conformation that favors corepressor binding, potentially antagonizing GRα function. This study thus presents an unexpected molecular mechanism by which GRβ could repress transcription.},
doi = {10.1128/MCB.00453-17},
journal = {Molecular and Cellular Biology},
number = 8,
volume = 38,
place = {United States},
year = {Mon Feb 05 00:00:00 EST 2018},
month = {Mon Feb 05 00:00:00 EST 2018}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 5 works
Citation information provided by
Web of Science
Web of Science
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Crystal Structure of the Glucocorticoid Receptor Ligand Binding Domain Reveals a Novel Mode of Receptor Dimerization and Coactivator Recognition
journal, July 2002
- Bledsoe, Randy K.; Montana, Valerie G.; Stanley, Thomas B.
- Cell, Vol. 110, Issue 1
Antiinflammatory Action of Glucocorticoids — New Mechanisms for Old Drugs
journal, October 2005
- Rhen, Turk; Cidlowski, John A.
- New England Journal of Medicine, Vol. 353, Issue 16
The Human Glucocorticoid Receptor Isoform: EXPRESSION, BIOCHEMICAL PROPERTIES, AND PUTATIVE FUNCTION
journal, April 1996
- Oakley, Robert H.; Sar, Madhabananda; Cidlowski, John A.
- Journal of Biological Chemistry, Vol. 271, Issue 16
The Physiology of Human Glucocorticoid Receptor beta (hGRbeta) and Glucocorticoid Resistance
journal, June 2006
- Lewis-Tuffin, L. J.
- Annals of the New York Academy of Sciences, Vol. 1069, Issue 1
Ligand-selective transactivation and transrepression via the glucocorticoid receptor: Role of cofactor interaction
journal, February 2009
- Ronacher, Katharina; Hadley, Katie; Avenant, Chanel
- Molecular and Cellular Endocrinology, Vol. 299, Issue 2
Increased number of glucocorticoid receptor-β–expressing cells in the airways in fatal asthma
journal, September 2000
- Christodoulopoulos, Pota; Leung, Donald Y. M.; Elliott, Mark W.
- Journal of Allergy and Clinical Immunology, Vol. 106, Issue 3
GRβ expression in nasal polyp inflammatory cells and its relationship to the anti-inflammatory effects of intranasal fluticasone☆
journal, July 2001
- Hamilos, Daniel L.; Leung, Donald Y. M.; Muro, Shegio
- Journal of Allergy and Clinical Immunology, Vol. 108, Issue 1
Isolation of a cDNA Encoding Human Rev-ErbAα: Transcription from the Noncoding DNA Strand of a Thyroid Hormone Receptor Gene Results in a Related Protein That Does Not Bind Thyroid Hormone
journal, March 1990
- Lazar, Mitchell A.; Jones, Karen E.; Chin, William W.
- DNA and Cell Biology, Vol. 9, Issue 2
Comparative Protein Structure Modeling Using Modeller
journal, September 2006
- Eswar, Narayanan; Webb, Ben; Marti-Renom, Marc A.
- Current Protocols in Bioinformatics, Vol. 15, Issue 1
DNA Binding Site Sequence Directs Glucocorticoid Receptor Structure and Activity
journal, April 2009
- Meijsing, S. H.; Pufall, M. A.; So, A. Y.
- Science, Vol. 324, Issue 5925
Imbalanced Expression of the Glucocorticoid Receptor Isoforms in Cultured Lymphocytes from a Patient with Systemic Glucocorticoid Resistance and Chronic Lymphocytic Leukemia
journal, January 1999
- Shahidi, Homayoun; Vottero, Alessandra; Stratakis, Constantine A.
- Biochemical and Biophysical Research Communications, Vol. 254, Issue 3
Significance of glucocorticoid receptor expression in colonic mucosal cells of patients with ulcerative colitis
journal, January 2005
- Zhang, Hu
- World Journal of Gastroenterology, Vol. 11, Issue 12
Widespread Negative Response Elements Mediate Direct Repression by Agonist- Liganded Glucocorticoid Receptor
journal, April 2011
- Surjit, Milan; Ganti, Krishna Priya; Mukherji, Atish
- Cell, Vol. 145, Issue 2
Expression of glucocorticoid receptor β in lymphocytes of patients with glucocorticoid-resistant ulcerative colitis
journal, May 2000
- Honda, Mitsunori; Orii, Fumika; Ayabe, Tokiyoshi
- Gastroenterology, Vol. 118, Issue 5
Nuclear Receptor Coregulators: Cellular and Molecular Biology*
journal, June 1999
- McKenna, Neil J.; Lanz, Rainer B.; O’Malley, Bert W.
- Endocrine Reviews, Vol. 20, Issue 3
MolProbity : all-atom structure validation for macromolecular crystallography
journal, December 2009
- Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1
Coot model-building tools for molecular graphics
journal, November 2004
- Emsley, Paul; Cowtan, Kevin
- Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
Molecular Switch in the Glucocorticoid Receptor: Active and Passive Antagonist Conformations
journal, January 2010
- Schoch, Guillaume A.; D'Arcy, Brigitte; Stihle, Martine
- Journal of Molecular Biology, Vol. 395, Issue 3
Gene regulation by the glucocorticoid receptor: Structure:function relationship
journal, April 2005
- Kumar, Raj; Thompson, E. Brad
- The Journal of Steroid Biochemistry and Molecular Biology, Vol. 94, Issue 5
Glucocorticoid receptor beta, a potential endogenous inhibitor of glucocorticoid action in humans.
journal, June 1995
- Bamberger, C. M.; Bamberger, A. M.; de Castro, M.
- Journal of Clinical Investigation, Vol. 95, Issue 6
The biology of the glucocorticoid receptor: New signaling mechanisms in health and disease
journal, November 2013
- Oakley, Robert H.; Cidlowski, John A.
- Journal of Allergy and Clinical Immunology, Vol. 132, Issue 5
PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
- Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
Association of Glucocorticoid Insensitivity with Increased Expression of Glucocorticoid Receptor
journal, November 1997
- Leung, D. Y. M.; Hamid, Q.; Vottero, A.
- Journal of Experimental Medicine, Vol. 186, Issue 9
Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions
journal, November 2004
- Krissinel, E.; Henrick, K.
- Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12
Translational Regulatory Mechanisms Generate N-Terminal Glucocorticoid Receptor Isoforms with Unique Transcriptional Target Genes
journal, April 2005
- Lu, Nick Z.; Cidlowski, John A.
- Molecular Cell, Vol. 18, Issue 3
Increased Glucocorticoid Receptor β in Airway Cells of Glucocorticoid-insensitive Asthma
journal, May 1999
- Hamid, Qutayba A.; Wenzel, Sally E.; Hauk, Pia J.
- American Journal of Respiratory and Critical Care Medicine, Vol. 159, Issue 5
Mechanisms involved in the side effects of glucocorticoids
journal, October 2002
- Schacke, H.
- Pharmacology & Therapeutics, Vol. 96, Issue 1
Low Glucocorticoid Receptor α/β Ratio in T-cell Lymphoblastic Leukemia
journal, October 2000
- Longui, C.; Vottero, A.; Adamson, P.
- Hormone and Metabolic Research, Vol. 32, Issue 10
Quantitative analysis for human glucocorticoid receptor α/β mRNA in IBD
journal, September 2002
- Orii, Fumika; Ashida, Toshifumi; Nomura, Masafumi
- Biochemical and Biophysical Research Communications, Vol. 296, Issue 5
Mifepristone Inhibits GRβ Coupled Prostate Cancer Cell Proliferation
journal, September 2012
- Ligr, Martin; Li, Yirong; Logan, Susan K.
- Journal of Urology, Vol. 188, Issue 3
Molecular Origins for the Dominant Negative Function of Human Glucocorticoid Receptor Beta
journal, June 2003
- Yudt, M. R.; Jewell, C. M.; Bienstock, R. J.
- Molecular and Cellular Biology, Vol. 23, Issue 12
Structure of Rev-erbα bound to N-CoR reveals a unique mechanism of nuclear receptor–co-repressor interaction
journal, June 2010
- Phelan, Caroline A.; Gampe, Robert T.; Lambert, Millard H.
- Nature Structural & Molecular Biology, Vol. 17, Issue 7
Expression and Subcellular Distribution of the β-Isoform of the Human Glucocorticoid Receptor 1
journal, November 1997
- Oakley, Robert H.; Webster, Jeffrey C.; Sar, Madhabananda
- Endocrinology, Vol. 138, Issue 11
RU486-induced Glucocorticoid Receptor Agonism Is Controlled by the Receptor N Terminus and by Corepressor Binding
journal, May 2002
- Schulz, Martin; Eggert, Martin; Baniahmad, Aria
- Journal of Biological Chemistry, Vol. 277, Issue 29
Primary structure and expression of a functional human glucocorticoid receptor cDNA
journal, December 1985
- Hollenberg, Stanley M.; Weinberger, Cary; Ong, Estelita S.
- Nature, Vol. 318, Issue 6047
Human Glucocorticoid Receptor β Regulates Gluconeogenesis and Inflammation in Mouse Liver
journal, December 2015
- He, Bo; Cruz-Topete, Diana; Oakley, Robert H.
- Molecular and Cellular Biology, Vol. 36, Issue 5
[20] Processing of X-ray diffraction data collected in oscillation mode
book, January 1997
- Otwinowski, Zbyszek; Minor, Wladek
- Macromolecular Crystallography Part A, 307-326
Human Glucocorticoid Receptor β Binds RU-486 and Is Transcriptionally Active
journal, January 2007
- Lewis-Tuffin, Laura J.; Jewell, Christine M.; Bienstock, Rachelle J.
- Molecular and Cellular Biology, Vol. 27, Issue 6
Conformational changes of the glucocorticoid receptor ligand binding domain induced by ligand and cofactor binding, and the location of cofactor binding sites determined by hydrogen/deuterium exchange mass spectrometry
journal, March 2006
- Frego, L.
- Protein Science, Vol. 15, Issue 4
Proinflammatory cytokines regulate human glucocorticoid receptor gene expression and lead to the accumulation of the dominant negative isoform: A mechanism for the generation of glucocorticoid resistance
journal, May 2001
- Webster, J. C.; Oakley, R. H.; Jewell, C. M.
- Proceedings of the National Academy of Sciences, Vol. 98, Issue 12
Relation between age, immunophenotype and in vitro drug resistance in 395 children with acute lymphoblastic leukemia–implications for treatment of infants
journal, September 1998
- Pieters, R.; den Boer, Ml; Durian, M.
- Leukemia, Vol. 12, Issue 9
Phaser crystallographic software
journal, July 2007
- McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
- Journal of Applied Crystallography, Vol. 40, Issue 4
The Orphan Nuclear Receptor Rev-erbα Recruits the N-CoR/Histone Deacetylase 3 Corepressor to Regulate the Circadian Bmal1 Gene
journal, June 2005
- Yin, Lei; Lazar, Mitchell A.
- Molecular Endocrinology, Vol. 19, Issue 6
Mechanisms of Corticosteroid Resistance in Rheumatoid Arthritis
journal, June 2002
- Chikanza, Ian C.
- Annals of the New York Academy of Sciences, Vol. 966, Issue 1
FRED Pose Prediction and Virtual Screening Accuracy
journal, February 2011
- McGann, Mark
- Journal of Chemical Information and Modeling, Vol. 51, Issue 3
Exploring the gas access routes in a [NiFeSe] hydrogenase using crystals pressurized with krypton and oxygen
journal, August 2020
- Zacarias, Sónia; Temporão, Adriana; Carpentier, Philippe
- JBIC Journal of Biological Inorganic Chemistry, Vol. 25, Issue 6
Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation
journal, February 2021
- Tillu, Vikas A.; Rae, James; Gao, Ya
- Nature Communications, Vol. 12, Issue 1
Human Glucocorticoid Receptor β Regulates Gluconeogenesis and Inflammation in Mouse Liver
text, January 2016
- A., Cidlowski, John; H., Oakley, Robert; Xiao, Xiao,
- The University of North Carolina at Chapel Hill University Libraries
PHENIX: a comprehensive Python-based system for macromolecular structure solution.
text, January 2010
- Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
- Apollo - University of Cambridge Repository
Works referencing / citing this record:
Clinical Use and Molecular Action of Corticosteroids in the Pediatric Age
journal, January 2019
- Ferrara, Giovanna; Petrillo, Maria; Giani, Teresa
- International Journal of Molecular Sciences, Vol. 20, Issue 2