title: Role of the proteasome in the regulation of estrogen receptor alpha turnover and function in MCF-7 breast carcinoma cells.
abstract: Estrogen receptor alpha (ER) turnover in MCF-7 cells was assessed by pulse chase analysis and measurement of ER steady-state level. In untreated cells, degradation of (35)S-labeled ER was characterized by a slow phase followed by a more rapid decline. Without ligand, ER elimination was totally compensated by synthesis which maintained receptor homeostasis. Estradiol (E(2)) and the pure antiestrogen RU 58,668 abolished the slow phase of ER breakdown and enhanced the degradation of neosynthesized ER, producing a low ER steady-state level. By contrast, the partial antiestrogen OH-Tam was ineffective in this respect and caused ER accumulation. Regardless of the conditions, ER breakdown was abolished by proteasome inhibition (MG-132). ER ligands decreased cell capacity to bind [(3)H]E(2), even in the presence of MG-132, indicating that the regulation of ER level and E(2) binding capacity occurs through distinct mechanisms. MG-132 partially blocked the basal transcription of an ERE-dependent reporter gene and modified the ability of E(2) to induce the expression of the latter: the hormone was unable to restore the transactivation activity measured without MG-132. RU 58,668 and OH-Tam failed to enhance the inhibitory action of MG-132, suggesting that a loss of basal ER-mediated transactivation mainly affects the stimulatory effect of estrogens. Overall, our findings reveal that ER steady state level, ligand binding capacity and transactivation potency fit in a complex regulatory scheme involving distinct mechanisms, which may be dissociated from each other under various treatments.
authors.affiliation: Laboratoire J.-C. Heuson de Cancérologie Mammaire, Service de Médecine, Institut Jules Bordet, Université Libre de Bruxelles, 1 rue Héger-Bordet, B-1000 Brussels, Belgium.
authors.forename: Ioanna
authors.initials: I
authors.lastname: Laïos
authors.forename: Fabrice
authors.initials: F
authors.lastname: Journé
authors.forename: Denis
authors.initials: D
authors.lastname: Nonclercq
authors.forename: Doris Salazar
authors.initials: DS
authors.lastname: Vidal
authors.forename: Robert-Alain
authors.initials: RA
authors.lastname: Toillon
authors.forename: Guy
authors.initials: G
authors.lastname: Laurent
authors.forename: Guy
authors.initials: G
authors.lastname: Leclercq
mesh_terms: D001943:Breast Neoplasms; D045744:Cell Line, Tumor; D003513:Cycloheximide / Q000494:pharmacology; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000187:drug effects / Q000378:metabolism*; D017930:Genes, Reporter; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D008715:Methionine; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D013462:Sulfur Radioisotopes; D013629:Tamoxifen / Q000031:analogs & derivatives* / Q000494:pharmacology; D014158:Transcription, Genetic; D014316:Tritium; D005466:Fluorocarbons
chemical_list: D003513:Cycloheximide; D004791:Enzyme Inhibitors; D004958:Estradiol; D047628:Estrogen Receptor alpha; D007976:Leupeptins; D008024:Ligands; D008156:Luciferases; D008715:Methionine; D046988:Proteasome Endopeptidase Complex; D061988:Proteasome Inhibitors; D013462:Sulfur Radioisotopes; D013629:Tamoxifen; D014316:Tritium; C086511:RU 58668; D005466:Fluorocarbons; C016601:afimoxifene; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde