Analysis of estrogen receptor interaction with a repressor of estrogen receptor activity (REA) and the regulation of estrogen receptor transcriptional activity by REA.
The transcriptional activity of nuclear hormone receptors is known to be modulated by coregulator proteins. We found that the repressor of estrogen receptor activity (REA), a protein recruited to the hormone-occupied estrogen receptor (ER), decreased the transcriptional activity of ER, both when ER was acting directly through DNA response elements as well as when it was tethered to other transcription factors. Administration of antisense REA resulted in a 2-4-fold increase in ER transactivation, implying that endogenous REA normally dampens the stimulatory response to estradiol. To define the interaction regions between ER and REA, we used glutathione S-transferase pull-down assays. We found that REA bound to the ligand-binding domain (E domain) of ER, but not to other regions of ER, and that REA interaction with ER involved a region in the C-terminal half of REA. REA and the coactivator SRC-1 were involved in a functional competition for regulation of ER transcriptional activity, which we show results from competition between these two coregulators for interaction with ER. REA contains an LXXLL motif near its N terminus, but this motif was not involved in its binding to ER. Rather, this sequence was required for the competitive binding of REA and SRC-1 to ER and thus for optimal repression of ER activity. Our findings show that the regions of REA required for its interaction with ER and for its repression of ER activity are different.
Delage-Mourroux
R
R
Departments of Molecular and Integrative Physiology and Cell and Structural Biology, University of Illinois and the College of Medicine, Urbana, Illinois 61801-3704, USA.
Martini
P G
PG
Choi
I
I
Kraichely
D M
DM
Hoeksema
J
J
Katzenellenbogen
B S
BS
D020816:Amino Acid Motifs; D000818:Animals; D001667:Binding, Competitive; D016384:Consensus Sequence / Q000235:genetics; D004958:Estradiol / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D051548:Histone Acetyltransferases; D006801:Humans; D009154:Mutation; D056920:Nuclear Receptor Coactivator 1; D016376:Oligonucleotides, Antisense / Q000235:genetics; D000091402:Prohibitins; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured
D016376:Oligonucleotides, Antisense; C000717331:PHB2 protein, human; D000091402:Prohibitins; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D004958:Estradiol; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1