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Molecular Endocrinology 14 (4): 518-534
Copyright © 2000 by The Endocrine Society

Subnuclear Trafficking of Estrogen Receptor-{alpha} and Steroid Receptor Coactivator-1

David L. Stenoien, Maureen G. Mancini, Kavita Patel, Elizabeth A. Allegretto*, Carolyn L. Smith and Michael A. Mancini

Department of Molecular and Cellular Biology (D.L.S., M.G.M., K.P., C.L.S., M.A.M.) Baylor College of Medicine Houston, Texas 77030
Ligand Pharmaceuticals, Inc. (E.A.A.) San Diego, California 92121

We have *analyzed ligand-dependent, subnuclear movements of the estrogen receptor-{alpha} (ER{alpha}) in terms of both spatial distribution and solubility partitioning. Using a transcriptionally active green fluorescent protein-ER{alpha} chimera (GFP-ER{alpha}), we find that 17ß-estradiol (E2) changes the normally diffuse nucleoplasmic pattern of GFP-ER{alpha} to a hyperspeckled distribution within 10–20 min. A similar reorganization occurs with the partial antagonist 4-hydroxytamoxifen; only a subtle effect was observed with the pure antagonist ICI 182,780. To examine the influence of ligand upon ER{alpha} association with nuclear structure, MCF-7 cells were extracted to reveal the nuclear matrix (NM). Addition of E2, 4-hydroxytamoxifen, or ICI 182,780 causes ER{alpha} to partition with the NM-bound fraction on a similar time course (10–20 min) as the spatial reorganization suggesting that the two events are related. To determine the effects of E2 on the redistribution and solubility of GFP-ER{alpha}, individual cells were directly examined during both hormone addition and NM extraction and showed that GFP-ER{alpha} movement and NM association were coincident. Colocalization experiments were performed with antibodies to identify sites of transcription (RNA pol IIo) and splicing domains (SRm160). Using E2 treated MCF-7 cells, minor overlap was observed with transcription sites and a small amount of the total ER{alpha} pool. Experiments performed with bioluminescent derivatives of ER{alpha} and steroid receptor coactivator-1 (SRC-1) demonstrated both proteins colocalize to the same NM-bound foci in response to E2 but not the antagonists tested. Deletion mutagenesis and in situ analyses indicate intranuclear colocalization requires a central SRC-1 domain containing LXXLL motifs. Collectively, our data suggest that ER{alpha} transcription function is dependent upon dynamic early events including intranuclear rearrangement, NM association, and SRC-1 interactions.




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J. Biol. Chem.Home page
D. J. Mulholland, H. Cheng, K. Reid, P. S. Rennie, and C. C. Nelson
The Androgen Receptor Can Promote beta -Catenin Nuclear Translocation Independently of Adenomatous Polyposis Coli
J. Biol. Chem., May 10, 2002; 277(20): 17933 - 17943.
[Abstract] [Full Text] [PDF]


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Hum Mol GenetHome page
D. J. Lloyd, R. C. Trembath, and S. Shackleton
A novel interaction between lamin A and SREBP1: implications for partial lipodystrophy and other laminopathies
Hum. Mol. Genet., April 1, 2002; 11(7): 769 - 777.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
M. Saitoh, R. Takayanagi, K. Goto, A. Fukamizu, A. Tomura, T. Yanase, and H. Nawata
The Presence of Both the Amino- and Carboxyl-Terminal Domains in the AR Is Essential for the Completion of a Transcriptionally Active Form with Coactivators and Intranuclear Compartmentalization Common to the Steroid Hormone Receptors: A Three-Dimensional Imaging Study
Mol. Endocrinol., April 1, 2002; 16(4): 694 - 706.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
T. E. Akiyama, C. T. Baumann, S. Sakai, G. L. Hager, and F. J. Gonzalez
Selective Intranuclear Redistribution of PPAR Isoforms by RXR{alpha}
Mol. Endocrinol., April 1, 2002; 16(4): 707 - 721.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
R. V. Weatherman, C.-Y. Chang, N. J. Clegg, D. C. Carroll, R. N. Day, J. D. Baxter, D. P. McDonnell, T. S. Scanlan, and F. Schaufele
Ligand-Selective Interactions of ER Detected in Living Cells by Fluorescence Resonance Energy Transfer
Mol. Endocrinol., March 1, 2002; 16(3): 487 - 496.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
R. AEsoy, G. Mellgren, K.-I. Morohashi, and J. Lund
Activation of cAMP-Dependent Protein Kinase Increases the Protein Level of Steroidogenic Factor-1
Endocrinology, January 1, 2002; 143(1): 295 - 303.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
F. Schaufele, J. F. Enwright III, X. Wang, C. Teoh, R. Srihari, R. Erickson, O. A. MacDougald, and R. N. Day
CCAAT/Enhancer Binding Protein {alpha} Assembles Essential Cooperating Factors in Common Subnuclear Domains
Mol. Endocrinol., October 1, 2001; 15(10): 1665 - 1676.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
K. L. Sunn, T.-A. Cock, L. A. Crofts, J. A. Eisman, and E. M. Gardiner
Novel N-Terminal Variant of Human VDR
Mol. Endocrinol., September 1, 2001; 15(9): 1599 - 1609.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
D. L. Stenoien, A. C. Nye, M. G. Mancini, K. Patel, M. Dutertre, B. W. O'Malley, C. L. Smith, A. S. Belmont, and M. A. Mancini
Ligand-Mediated Assembly and Real-Time Cellular Dynamics of Estrogen Receptor {alpha}-Coactivator Complexes in Living Cells
Mol. Cell. Biol., July 1, 2001; 21(13): 4404 - 4412.
[Abstract] [Full Text] [PDF]


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Physiol. Rev.Home page
P. M. Yen
Physiological and Molecular Basis of Thyroid Hormone Action
Physiol Rev, July 1, 2001; 81(3): 1097 - 1142.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J.-Y. Choi, J. Pratap, A. Javed, S. K. Zaidi, L. Xing, E. Balint, S. Dalamangas, B. Boyce, A. J. van Wijnen, J. B. Lian, et al.
Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development
PNAS, June 28, 2001; (2001) 151236498.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
C. Pasqualini, D. Guivarc'h, J.-V. Barnier, B. Guibert, J.-D. Vincent, and P. Vernier
Differential Subcellular Distribution and Transcriptional Activity of {{Sigma}}E3, {{Sigma}}E4, and {{Sigma}}E3-4 Isoforms of the Rat Estrogen Receptor-{{alpha}}
Mol. Endocrinol., June 1, 2001; 15(6): 894 - 908.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
R. H. Price Jr., C. A. Butler, P. Webb, R. Uht, P. Kushner, and R. J. Handa
A Splice Variant of Estrogen Receptor {beta} Missing Exon 3 Displays Altered Subnuclear Localization and Capacity for Transcriptional Activation
Endocrinology, May 1, 2001; 142(5): 2039 - 2049.
[Abstract] [Full Text]


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J. Cell Sci.Home page
C. Herrmann and M. Mancini
The Cdk9 and cyclin T subunits of TAK/P-TEFb localize to splicing factor-rich nuclear speckle regions
J. Cell Sci., January 4, 2001; 114(8): 1491 - 1503.
[Abstract] [PDF]


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J. Cell Sci.Home page
J Nickerson
Experimental observations of a nuclear matrix
J. Cell Sci., January 2, 2001; 114(3): 463 - 474.
[Abstract] [PDF]


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Mol. Endocrinol.Home page
F. Schaufele, C.-y. Chang, W. Liu, J. D. Baxter, S. K. Nordeen, Y. Wan, R. N. Day, and D. P. McDonnell
Temporally Distinct and Ligand-Specific Recruitment of Nuclear Receptor-Interacting Peptides and Cofactors to Subnuclear Domains Containing the Estrogen Receptor
Mol. Endocrinol., December 1, 2000; 14(12): 2024 - 2039.
[Abstract] [Full Text]


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Mol. Endocrinol.Home page
R. K. Tyagi, Y. Lavrovsky, S. C. Ahn, C. S. Song, B. Chatterjee, and A. K. Roy
Dynamics of Intracellular Movement and Nucleocytoplasmic Recycling of the Ligand-Activated Androgen Receptor in Living Cells
Mol. Endocrinol., August 1, 2000; 14(8): 1162 - 1174.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
K. Prufer, A. Racz, G. C. Lin, and J. Barsony
Dimerization with Retinoid X Receptors Promotes Nuclear Localization and Subnuclear Targeting of Vitamin D Receptors
J. Biol. Chem., December 22, 2000; 275(52): 41114 - 41123.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
C. T. Baumann, P. Maruvada, G. L. Hager, and P. M. Yen
Nuclear Cytoplasmic Shuttling by Thyroid Hormone Receptors. MULTIPLE PROTEIN INTERACTIONS ARE REQUIRED FOR NUCLEAR RETENTION
J. Biol. Chem., March 30, 2001; 276(14): 11237 - 11245.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
A. Tomura, K. Goto, H. Morinaga, M. Nomura, T. Okabe, T. Yanase, R. Takayanagi, and H. Nawata
The Subnuclear Three-dimensional Image Analysis of Androgen Receptor Fused to Green Fluorescence Protein
J. Biol. Chem., July 20, 2001; 276(30): 28395 - 28401.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J.-M. Sun, H. Y. Chen, and J. R. Davie
Effect of Estradiol on Histone Acetylation Dynamics in Human Breast Cancer Cells
J. Biol. Chem., December 21, 2001; 276(52): 49435 - 49442.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J.-Y. Choi, J. Pratap, A. Javed, S. K. Zaidi, L. Xing, E. Balint, S. Dalamangas, B. Boyce, A. J. van Wijnen, J. B. Lian, et al.
Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development
PNAS, July 17, 2001; 98(15): 8650 - 8655.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Endocrinol. Metab.Home page
M. T. Preisler-Mashek, N. Solodin, B. L. Stark, M. K. Tyriver, and E. T. Alarid
Ligand-specific regulation of proteasome-mediated proteolysis of estrogen receptor-alpha
Am J Physiol Endocrinol Metab, April 1, 2002; 282(4): E891 - E898.
[Abstract] [Full Text] [PDF]




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