Proteasome-mediated proteolysis of estrogen receptor: a novel component in autologous down-regulation.
Regulation of estrogen receptor (ER) concentration is a key component in limiting estrogen responsiveness in target cells. Yet the mechanisms governing ER concentration in the lactotrope cells of the anterior pituitary, a major site of estrogen action, are undetermined. In this study, we used a lactotrope cell line, PR1, to explore regulation of ER protein by estrogen. Estrogen treatment resulted in an approximate 60% decrease in ER steady state protein levels. Suprisingly, the decline in ER protein was apparent within 1 h of estrogen treatment and occurred in the absence of protein synthesis and transcription. Direct regulation of ER protein was further confirmed by pulse chase analysis, which showed that ER protein half-life was shortened from greater than 3 h to 1 h in the presence of estrogen. The estrogen-induced degradation of ER protein could be prevented by pretreatment with peptide aldehyde inhibitors of proteasome protease whereas inhibitors of calpain and lysosomal proteases were ineffective. Inhibition of proteasome activity maintained ER protein at a level equivalent to control cells not stimulated with estrogen but increased estrogen-binding activity by 1.75-fold. Proteolytic regulation of ER by the proteasome is not limited to pituitary lactotrope cells but is also operational in MCF-7 breast cancer cells, suggesting that this may be a common regulatory pathway used by estrogen. These studies describe a nongenomic action of estrogen that involves nuclear ER: rapid proteolysis of ER protein via a proteasome-mediated pathway.
Alarid
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The Department of Physiology, University of Wisconsin-Madison, 53706, USA. alarid@physiology.wisc.edu
Bakopoulos
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Solodin
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D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002461:Cell Line, Transformed; D003546:Cysteine Endopeptidases / Q000502:physiology*; D015536:Down-Regulation / Q000187:drug effects; D004967:Estrogens / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D009097:Multienzyme Complexes / Q000502:physiology*; D010902:Pituitary Gland / Q000166:cytology; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004967:Estrogens; D009097:Multienzyme Complexes; D011480:Protease Inhibitors; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex