|
Originally published In Press as doi:10.1074/jbc.M306199200 on August 12, 2003
J. Biol. Chem., Vol. 278, Issue 44, 43797-43806, October 31, 2003
Molecular Determinants of the Balance between Co-repressor and Co-activator Recruitment to the Retinoic Acid Receptor*
Szilvia Benko ,
James D. Love ,
Marta Beládi,
John W. R. Schwabe¶, and
Laszlo Nagy¶||
From the
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen, Medical and Health Science Center, Nagyerdei krt. 98, Debrecen H-4012, Hungary and the Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom
The repressive and activating states of nuclear hormone receptors are achieved through the recruitment of cofactor proteins. The binding of co-repressors and co-activators is believed to be mutually exclusive and principally regulated by ligand binding. To understand the molecular determinants of the switch induced by ligand in the retinoic acid receptor and in particular the intrinsic role of the ligand binding domain (LBD) in cofactor binding and release, we carried out extensive mutational analysis of surface residues of the LBD. As seen previously we found that co-repressor and co-activator molecules bind to overlapping docking sites on the surface of the retinoic acid receptor  LBD. Perturbation of this surface impaired both co-activator and co-repressor association resulting in a transcriptionally inert receptor. Unexpectedly mutation of two residues, Trp-225 and Ala-392, which lie outside the docking site, had opposite effects on co-activator and co-repressor binding. W225A was a constitutive repressor that failed to bind co-activator and exhibited an increased, and ligand-insensitive, interaction with co-repressor. A392R, on the other hand, had reduced affinity for co-repressors and increased affinity for co-activators and behaved as a constitutive, but still ligand-inducible, activator. Analysis of known structures showed that these mutations lie in the proximity of helix 12 (H12), and their effects are likely to be the result of perturbations in the behavior of H12. These data suggest that residues in the close vicinity of H12 regulate cofactor affinity and determine the basal activity of receptors.
Received for publication, June 12, 2003
, and in revised form, August 6, 2003.
* This work was supported by grants from the Royal Society, Human Frontier Science Program, and Research Training Network from the European Union FP5 (to L. N. and J. W. R. S.), a research award from the Boehringer Ingelheim Fund (to L. N.), and Grant T034434 from the Hungarian Scientific Research Fund (to L. N.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
¶ Both authors share senior authorship.
|| An international scholar of the Howard Hughes Medical Research Institute and a European Molecular Biology Organization young investigator. To whom correspondence should be addressed. Tel.: 36-52-416-432; Fax: 36-52-314-989; E-mail: lnagy{at}indi.biochem.dote.hu.
 CiteULike  Complore  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
G. Pochetti, C. Godio, N. Mitro, D. Caruso, A. Galmozzi, S. Scurati, F. Loiodice, G. Fracchiolla, P. Tortorella, A. Laghezza, et al.
Insights into the Mechanism of Partial Agonism: CRYSTAL STRUCTURES OF THE PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR {gamma} LIGAND-BINDING DOMAIN IN THE COMPLEX WITH TWO ENANTIOMERIC LIGANDS
J. Biol. Chem.,
June 8, 2007;
282(23):
17314 - 17324.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Szatmari, G. Vamosi, P. Brazda, B. L. Balint, S. Benko, L. Szeles, V. Jeney, C. Ozvegy-Laczka, A. Szanto, E. Barta, et al.
Peroxisome Proliferator-activated Receptor {gamma}-regulated ABCG2 Expression Confers Cytoprotection to Human Dendritic Cells
J. Biol. Chem.,
August 18, 2006;
281(33):
23812 - 23823.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. H. Kim, M. H. Lee, B. J. Kim, J. H. Kim, S. J. Han, H. Y. Kim, and M. R Stallcup
Role of aspartate 351 in transactivation and active conformation of estrogen receptor {alpha}
J. Mol. Endocrinol.,
December 1, 2005;
35(3):
449 - 464.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. L. Balint, A. Szanto, A. Madi, U.-M. Bauer, P. Gabor, S. Benko, L. G. Puskas, P. J. A. Davies, and L. Nagy
Arginine Methylation Provides Epigenetic Transcription Memory for Retinoid-Induced Differentiation in Myeloid Cells
Mol. Cell. Biol.,
July 1, 2005;
25(13):
5648 - 5663.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Zechel
Requirement of Retinoic Acid Receptor Isotypes {alpha}, {beta}, and {gamma} during the Initial Steps of Neural Differentiation of PCC7 Cells
Mol. Endocrinol.,
June 1, 2005;
19(6):
1629 - 1645.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Jyrkkarinne, B. Windshugel, J. Makinen, M. Ylisirnio, M. Perakyla, A. Poso, W. Sippl, and P. Honkakoski
Amino Acids Important for Ligand Specificity of the Human Constitutive Androstane Receptor
J. Biol. Chem.,
February 18, 2005;
280(7):
5960 - 5971.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Codina, G. Benoit, J. T. Gooch, D. Neuhaus, T. Perlmann, and J. W. R. Schwabe
Identification of a Novel Co-regulator Interaction Surface on the Ligand Binding Domain of Nurr1 Using NMR Footprinting
J. Biol. Chem.,
December 17, 2004;
279(51):
53338 - 53345.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Szanto, S. Benko, I. Szatmari, B. L. Balint, I. Furtos, R. Ruhl, S. Molnar, L. Csiba, R. Garuti, S. Calandra, et al.
Transcriptional Regulation of Human CYP27 Integrates Retinoid, Peroxisome Proliferator-Activated Receptor, and Liver X Receptor Signaling in Macrophages
Mol. Cell. Biol.,
September 15, 2004;
24(18):
8154 - 8166.
[Abstract]
[Full Text]
[PDF]
|
|
|
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|
