4-Hydroxytamoxifen binds to and deactivates the estrogen-related receptor γ

The estrogen-related receptors (ERRα, ERRβ, and ERRγ) form a family of orphan nuclear receptors that share significant amino acid identity with the estrogen receptors, but for which physiologic roles remain largely unknown. By using a peptide sensor assay, we have identified the stilbenes diethylstilbestrol (DES), tamoxifen (TAM), and 4-hydroxytamoxifen (4-OHT) as high-affinity ligands for ERRγ. In direct binding assays, 4-OHT had a Kd value of 35 nM, and both DES and TAM displaced radiolabeled 4-OHT with Ki values of 870 nM. In cell-based assays, 4-OHT binding caused a dissociation of the complex between ERRγ and the steroid receptor coactivator-1, and led to an inhibition of the constitutive transcriptional activity of ERRγ. ERRα did not bind 4-OHT, but replacing a single amino acid predicted to be in the ERRα ligand-binding pocket with the corresponding ERRγ residue allowed high-affinity 4-OHT binding. These results demonstrate the existence of high-affinity ligands for the ERR family of orphan receptors, and identify 4-OHT as a molecule that can regulate the transcriptional activity of ERRγ.

[1]  C. Glass,et al.  Coactivator and corepressor complexes in nuclear receptor function. , 1999, Current opinion in genetics & development.

[2]  D. J. Heard,et al.  Human ERRgamma, a third member of the estrogen receptor-related receptor (ERR) subfamily of orphan nuclear receptors: tissue-specific isoforms are expressed during development and in the adult. , 2000, Molecular endocrinology.

[3]  R. Evans,et al.  Constitutive activation of transcription and binding of coactivator by estrogen-related receptors 1 and 2. , 1999, Molecular endocrinology.

[4]  J. Gustafsson,et al.  Transcriptional targets shared by estrogen receptor‐related receptors (ERRs) and estrogen receptor (ER) α, but not by ERβ , 1999 .

[5]  Shiuan Chen,et al.  Modulation of Aromatase Expression in the Breast Tissue by ERRα-1 Orphan Receptor , 1998 .

[6]  Gerhart U. Ryffel,et al.  An estrogen-responsive element derived from the 5′ flanking region of the Xenopus vitellogenin A2 gene functions in transfected human cells , 1986, Cell.

[7]  David M. Heery,et al.  A signature motif in transcriptional co-activators mediates binding to nuclear receptors , 1997, Nature.

[8]  K. Grandien,et al.  Printed in U.S.A. Copyright © 1997 by The Endocrine Society Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors � and � , 2022 .

[9]  S. Blanchard,et al.  Development of a scintillation proximity assay for peroxisome proliferator-activated receptor gamma ligand binding domain. , 1998, Analytical biochemistry.

[10]  J. Rossant,et al.  Placental abnormalities in mouse embryos lacking the orphan nuclear receptor ERR-β , 1997, Nature.

[11]  C. Teng,et al.  Estrogen Receptor-related Receptor α1 Interacts with Coactivator and Constitutively Activates the Estrogen Response Elements of the Human Lactoferrin Gene* , 2000, The Journal of Biological Chemistry.

[12]  J. Lehmann,et al.  Bile acids: natural ligands for an orphan nuclear receptor. , 1999, Science.

[13]  D. Mangelsdorf,et al.  An oxysterol signalling pathway mediated by the nuclear receptor LXRα , 1996, Nature.

[14]  D. Waxman,et al.  P450 gene induction by structurally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. , 1999, Archives of biochemistry and biophysics.

[15]  M. Makishima,et al.  Identification of a nuclear receptor for bile acids. , 1999, Science.

[16]  V. Laudet,et al.  Activation of the thyroid hormone receptor α gene promoter by the orphan nuclear receptor ERRα , 1998, Oncogene.

[17]  V. Laudet,et al.  Activation of the osteopontin promoter by the orphan nuclear receptor estrogen receptor related alpha. , 1998, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[18]  G. Zhou,et al.  Nuclear receptors have distinct affinities for coactivators: characterization by fluorescence resonance energy transfer. , 1998, Molecular endocrinology.

[19]  R. Evans,et al.  Identification of a new class of steroid hormone receptors , 1988, Nature.

[20]  David A. Agard,et al.  The Structural Basis of Estrogen Receptor/Coactivator Recognition and the Antagonism of This Interaction by Tamoxifen , 1998, Cell.

[21]  C. Yang,et al.  Two organochlorine pesticides, toxaphene and chlordane, are antagonists for estrogen-related receptor alpha-1 orphan receptor. , 1999, Cancer research.

[22]  T. Willson,et al.  The PPARs: from orphan receptors to drug discovery. , 2000, Journal of medicinal chemistry.

[23]  Timothy M. Willson,et al.  Activation of the Nuclear Receptor LXR by Oxysterols Defines a New Hormone Response Pathway* , 1997, The Journal of Biological Chemistry.

[24]  M. Stallcup,et al.  Hormone-independent Transcriptional Activation and Coactivator Binding by Novel Orphan Nuclear Receptor ERR3* , 1999, The Journal of Biological Chemistry.

[25]  J. Rossant,et al.  Diethylstilbestrol regulates trophoblast stem cell differentiation as a ligand of orphan nuclear receptor ERR beta. , 2001, Genes & development.

[26]  C. Teng,et al.  Estrogen-related Receptor, hERR1, Modulates Estrogen Receptor-mediated Response of Human Lactoferrin Gene Promoter (*) , 1996, The Journal of Biological Chemistry.

[27]  L. Freedman.  Increasing the Complexity of Coactivation in Nuclear Receptor Signaling , 1999, Cell.

[28]  V. Giguère,et al.  The orphan nuclear receptor estrogen-related receptor alpha is a transcriptional regulator of the human medium-chain acyl coenzyme A dehydrogenase gene , 1997, Molecular and cellular biology.

[29]  L. Moore,et al.  Orphan Nuclear Receptors Constitutive Androstane Receptor and Pregnane X Receptor Share Xenobiotic and Steroid Ligands* , 2000, The Journal of Biological Chemistry.

[30]  Rick B. Vega,et al.  A Role for Estrogen-related Receptor α in the Control of Mitochondrial Fatty Acid β-Oxidation during Brown Adipocyte Differentiation* , 1997, The Journal of Biological Chemistry.

[31]  V. Laudet,et al.  Transcriptional Activities of the Orphan Nuclear Receptor ERRα (Estrogen Receptor-Related Receptor-α) , 1999 .