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Mol Cell Biol, March 1998, p. 1369-1378, Vol. 18, No. 3
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Nuclear Corepressors NCoR and SMRT Are Key
Regulators of Both Ligand- and 8-Bromo-Cyclic AMP-Dependent
Transcriptional Activity of the Human Progesterone Receptor
Brandee L.
Wagner,1
John D.
Norris,1
Trina A.
Knotts,2
Nancy L.
Weigel,2 and
Donald P.
McDonnell1,*
Department of Pharmacology and Cancer
Biology, Duke University Medical Center, Durham, North Carolina
27710,1 and
Department of Cell Biology,
Baylor College of Medicine, Houston, Texas 770302
Received 4 August 1997/Returned for modification 9 September
1997/Accepted 3 December 1997
Previously, we defined a novel class of ligands for the human
progesterone receptor (PR) which function as mixed agonists. These
compounds induce a conformational change upon binding the receptor that
is different from those induced by agonists and antagonists. This
establishes a correlation between the structure of a ligand-receptor
complex and its transcriptional activity. In an attempt to define the
cellular components which distinguish between different ligand-induced
PR conformations, we have determined, by using a mammalian two-hybrid
assay, that the nuclear receptor corepressor (NCoR) and the silencing
mediator for retinoid and thyroid hormone receptor (SMRT)
differentially associate with PR depending upon the class of ligand
bound to the receptor. Specifically, we observed that the corepressors
preferentially associate with antagonist-occupied PR and that
overexpression of these corepressors suppresses the partial agonist
activity of antagonist-occupied PR. Binding studies performed in vitro,
however, reveal that recombinant SMRT can interact with PR in a manner
which is not influenced by the nature of the bound ligand. Thus, the
inability of SMRT or NCoR to interact with agonist-activated PR when
assayed in vivo may relate more to the increased affinity of PR for
coactivators, with a subsequent displacement of corepressors, than to
an inherent low affinity for the corepressor proteins. Previous work
from other groups has shown that 8-bromo-cyclic AMP (8-bromo-cAMP) can
convert the PR antagonist RU486 into an agonist and, additionally, can
potentiate the transcriptional activity of agonist-bound PR. In this
study, we show that exogenous expression of NCoR or SMRT suppresses all
8-bromo-cAMP-mediated potentiation of PR transcriptional activity.
Further analysis revealed that 8-bromo-cAMP addition decreases the
association of NCoR and SMRT with PR. Thus, we propose that
8-bromo-cAMP-mediated potentiation of PR transcriptional activity is
due, at least in part, to a disruption of the interaction between PR
and the corepressors NCoR and SMRT. Cumulatively, these results suggest
that NCoR and SMRT expression may play a pivotal role in PR
pharmacology.
*
Corresponding author. Mailing address: Department of
Pharmacology and Cancer Biology, Duke University Medical Center, P.O. Box 3813, Durham, NC 27710. Phone: (919) 684-6035. Fax: (919) 681-7139. E-mail: mcdon016{at}acpub.duke.edu.
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