title: A conserved lysine in the estrogen receptor DNA binding domain regulates ligand activation profiles at AP-1 sites, possibly by controlling interactions with a modulating repressor.
abstract: BACKGROUND: Estrogen receptors alpha and beta (ERalpha and ERbeta) differentially activate genes with AP-1 elements. ERalpha activates AP-1 targets via activation functions with estrogens (the AF-dependent pathway), whereas ERbeta, and a short version of ERalpha (ERalpha DBD-LBD) activate only with anti-estrogens (AF-independent pathway). The DNA binding domain (DBD) plays an important role in both pathways, even though neither pathway requires ERE recognition. RESULTS: Mutations of a highly conserved DBD lysine (ERalpha.K206A/G), lead to super-activation of AP-1 through activation function dependent pathways, up to 200 fold. This super-activity can be elicited either through ER AFs 1 or 2, or that of a heterologous activation function (VP16). The homologous substitution in ERbeta, K170A, or in ERalpha DBD-LBD leads to estrogen-dependent AP-1 activation and loss of the usually potent anti-estrogen effects. Each of numerous K206 substitutions in ERalpha, except K206R, eliminates anti-estrogen activation and this loss correlates perfectly with a loss of ability to titrate a repressive function from the RU486 bound progesterone receptor. CONCLUSION: We conclude that ER DBDs contain a complex regulatory function that influences ligand activation profiles at AP-1. This function, which requires the integrity of the conserved lysine, both allows for activation at AP-1 with anti-estrogens (with ERbeta and ERalpha DBD-LBD), and prevents ERalpha from becoming superactive at AP-1 with estrogens. We discuss the possibility that a repressor interaction with the DBD both mediates the AF-independent pathway and dampens the AF dependent pathway. Mutations in the conserved lysine might, by this model, disrupt the binding or function of the repressor.
authors.affiliation: Departments of Pathology, & Biochemistry and Molecular Genetics University of Virginia, School of Medicine, MR5 Rm, 3123, 415 Lane Rd, Charlottesville, VA 22908-0904, USA. ruht@virginia.edu
authors.forename: Rosalie M
authors.initials: RM
authors.lastname: Uht
authors.forename: Paul
authors.initials: P
authors.lastname: Webb
authors.forename: Phuong
authors.initials: P
authors.lastname: Nguyen
authors.forename: Richard H
authors.initials: RH
authors.lastname: Price
authors.forename: Cathleen
authors.initials: C
authors.lastname: Valentine
authors.forename: Helene
authors.initials: H
authors.lastname: Favre
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Kushner