help button home button Endocrine Society Molecular Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Weatherman, R. V.
Right arrow Articles by Schaufele, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Weatherman, R. V.
Right arrow Articles by Schaufele, F.
Molecular Endocrinology 16 (3): 487-496
Copyright © 2002 by The Endocrine Society

Ligand-Selective Interactions of ER Detected in Living Cells by Fluorescence Resonance Energy Transfer

R. V. Weatherman1, C.-Y. Chang, N. J. Clegg, D. C. Carroll, R. N. Day, J. D. Baxter, D. P. McDonnell, T. S. Scanlan and F. Schaufele

Departments of Pharmaceutical Chemistry and Cellular & Molecular Pharmacology (R.V.W., N.J.C., D.C.C., T.S.S.), University of California, San Francisco, California 94143-0446; Department of Pharmacology and Cancer Biology (C.-Y.C., D.P.M.), Duke University Medical Center, Durham, North Carolina 27710; Graduate Program in Chemistry and Chemical Biology (N.J.C.), University of California, San Francisco, California 94143-0446; Departments of Medicine and Cell Biology (R.N.D.), NSF Center for Biological Timing, University of Virginia Health Sciences Center, Charlottesville, Virginia, 22908; Metabolic Research Unit, Diabetes Center and Department of Medicine (J.D.B., F.S.), University of California, San Francisco, California 94143-0540

Address all correspondence and requests for reprints to: Fred Schaufele, HSW-1119, 513 Parnassus, University of California, San Francisco, California 94143-0540

Some aspects of ligand-regulated transcription activation by the estrogen receptor (ER) are associated with the estrogen-dependent formation of a hydrophobic cleft on the receptor surface. At least in vitro, this cleft is required for direct interaction of ER with an {alpha} helix, containing variants of the sequence LXXLL, found in many coactivators. In cells, it is unknown whether ER interactions with the different LXXLL-containing helices are uniformly similar or whether they vary with LXXLL sequence or activating ligand. Using fluorescence resonance energy transfer (FRET), we confirm in the physiological environment a direct interaction between the estradiol (E2)-bound ER and LXXLL peptides expressed in living cells as fusions with spectral variants of the green fluorescent protein. This interaction was blocked by a single amino acid mutation in the hydrophobic cleft. No FRET was detected when cells were incubated with the antiestrogenic ligands tamoxifen and ICI 182,780. E2, diethylstilbestrol, ethyl indenestrol A, and 6,4'-dihydroxyflavone all promoted FRET and activated ER-dependent transcription. Measurement of the level of FRET of ER with different LXXLL-containing peptides suggested that the orientations or affinities of the LXXLL interactions with the hydrophobic cleft were globally similar but slightly different for some activating ligands.




This article has been cited by other articles:


Home page
 J. Neurosci.Home page
S. M. Fernandez, M. C. Lewis, A. S. Pechenino, L. L. Harburger, P. T. Orr, J. E. Gresack, G. E. Schafe, and K. M. Frick
Estradiol-Induced Enhancement of Object Memory Consolidation Involves Hippocampal Extracellular Signal-Regulated Kinase Activation and Membrane-Bound Estrogen Receptors
J. Neurosci., August 27, 2008; 28(35): 8660 - 8667.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Sanchez, K. Sauve, N. Picard, and A. Tremblay
The Hormonal Response of Estrogen Receptor beta Is Decreased by the Phosphatidylinositol 3-Kinase/Akt Pathway via a Phosphorylation-dependent Release of CREB-binding Protein
J. Biol. Chem., February 16, 2007; 282(7): 4830 - 4840.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
A. Padron, L. Li, E. M. Kofoed, and F. Schaufele
Ligand-Selective Interdomain Conformations of Estrogen Receptor-{alpha}
Mol. Endocrinol., January 1, 2007; 21(1): 49 - 61.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
R. Paulmurugan and S. S. Gambhir
An intramolecular folding sensor for imaging estrogen receptor-ligand interactions
PNAS, October 24, 2006; 103(43): 15883 - 15888.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
S. Khan, R. Barhoumi, R. Burghardt, S. Liu, K. Kim, and S. Safe
Molecular Mechanism of Inhibitory Aryl Hydrocarbon Receptor--Estrogen Receptor/Sp1 Cross Talk in Breast Cancer Cells
Mol. Endocrinol., September 1, 2006; 20(9): 2199 - 2214.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
F. Schaufele, X. Carbonell, M. Guerbadot, S. Borngraeber, M. S. Chapman, A. A. K. Ma, J. N. Miner, and M. I. Diamond
The structural basis of androgen receptor activation: Intramolecular and intermolecular amino-carboxy interactions
PNAS, July 12, 2005; 102(28): 9802 - 9807.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
R. N. Day and F. Schaufele
Imaging Molecular Interactions in Living Cells
Mol. Endocrinol., July 1, 2005; 19(7): 1675 - 1686.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
K. Kim, R. Barhoumi, R. Burghardt, and S. Safe
Analysis of Estrogen Receptor {alpha}-Sp1 Interactions in Breast Cancer Cells by Fluorescence Resonance Energy Transfer
Mol. Endocrinol., April 1, 2005; 19(4): 843 - 854.
[Abstract] [Full Text] [PDF]

Home page
 J Mol EndocrinolHome page
J. S Lewis, T J Thomas, R. G Pestell, C. Albanese, M. A Gallo, and T. Thomas
Differential effects of 16{alpha}-hydroxyestrone and 2-methoxyestradiol on cyclin D1 involving the transcription factor ATF-2 in MCF-7 breast cancer cells
J. Mol. Endocrinol., February 1, 2005; 34(1): 91 - 105.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
S. E. Ross, H. S. Radomska, B. Wu, P. Zhang, J. N. Winnay, L. Bajnok, W. S. Wright, F. Schaufele, D. G. Tenen, and O. A. MacDougald
Phosphorylation of C/EBP{alpha} Inhibits Granulopoiesis
Mol. Cell. Biol., January 15, 2004; 24(2): 675 - 686.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
M. Tanaka, M. Nishi, M. Morimoto, T. Sugimoto, and M. Kawata
Yellow Fluorescent Protein-Tagged and Cyan Fluorescent Protein-Tagged Imaging Analysis of Glucocorticoid Receptor and Importins in Single Living Cells
Endocrinology, September 1, 2003; 144(9): 4070 - 4079.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
Y. Bai and V. Giguere
Isoform-Selective Interactions between Estrogen Receptors and Steroid Receptor Coactivators Promoted by Estradiol and ErbB-2 Signaling in Living Cells
Mol. Endocrinol., April 1, 2003; 17(4): 589 - 599.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
F. Schaufele, X. Wang, X. Liu, and R. N. Day
Conformation of CCAAT/Enhancer-binding Protein alpha Dimers Varies with Intranuclear Location in Living Cells
J. Biol. Chem., March 14, 2003; 278(12): 10578 - 10587.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
R. N. Day, T. C. Voss, J. F. Enwright III, C. F. Booker, A. Periasamy, and F. Schaufele
Imaging the Localized Protein Interactions Between Pit-1 and the CCAAT/Enhancer Binding Protein {alpha} in the Living Pituitary Cell Nucleus
Mol. Endocrinol., March 1, 2003; 17(3): 333 - 345.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals
Copyright © 2002 by The Endocrine Society