Cyclic, proteasome-mediated turnover of unliganded and liganded ERalpha on responsive promoters is an integral feature of estrogen signaling.
We present an integrated model of hERalpha-mediated transcription where both unliganded and liganded receptors cycle on estrogen-responsive promoters. Using ChIP, FRAP, and biochemical analysis we evaluate hERalpha at several points in these cycles, establishing the ubiquitination status and subnuclear distribution of hERalpha, its mobility, the kinetics of transcriptional activation, and the cyclic recruitment of E3 ligases and the 19S regulatory component of the proteasome. These experiments, together with an evaluation of the inhibition of transcription and proteasome action, demonstrate that proteasome-mediated degradation and hERalpha-mediated transactivation are inherently linked and act to continuously turn over hERalpha on responsive promoters. Cyclic turnover of hERalpha permits continuous responses to changes in the concentration of estradiol.
Reid
George
G
European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117, Heidelberg, Germany.
Hübner
Michael R
MR
Métivier
Raphaël
R
Brand
Heike
H
Denger
Stefanie
S
Manu
Dominique
D
Beaudouin
Joël
J
Ellenberg
Jan
J
Gannon
Frank
F
D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000378:metabolism; D047628:Estrogen Receptor alpha; D004967:Estrogens / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D009097:Multienzyme Complexes / Q000378:metabolism; D009841:Oligonucleotides / Q000494:pharmacology; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured
D002843:Chromatin; D047628:Estrogen Receptor alpha; D004967:Estrogens; D008024:Ligands; D009097:Multienzyme Complexes; D009841:Oligonucleotides; D011960:Receptors, Estrogen; D004958:Estradiol; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex