title: Evidence for a common step in three different processes for modulating the kinetic properties of glucocorticoid receptor-induced gene transcription.
abstract: The dose-response curve of steroid hormones and the associated EC(50) value are critical parameters both in the development of new pharmacologically active compounds and in the endocrine therapy of various disease states. We have recently described three different variables that can reposition the dose-response curve of agonist-bound glucocorticoid receptors (GRs): a 21-base pair sequence of the rat tyrosine aminotransferase gene called a glucocorticoid modulatory element (GME), GR concentration, and coactivator concentration. At the same time, each of these three components was found to influence the partial agonist activity of antiglucocorticoids. In an effort to determine whether these three processes proceed via independent pathways or a common intermediate, we have examined several mechanistic details. The effects of increasing concentrations of both GR and the coactivator TIF2 are found to be saturable. Furthermore, saturating levels of either GR or TIF2 inhibit the ability of each protein, and the GME, to affect further changes in the dose-response curve or partial agonist activity of antisteroids. This competitive inhibition suggests that all three modulators proceed through a common step involving a titratable factor. Support for this hypothesis comes from the observation that a fragment of the coactivator TIF2 retaining intrinsic transactivation activity is a dominant negative inhibitor of each component (GME, GR, and coactivator). This inhibition was not due to nonspecific effects on the general transcription machinery as the VP16 transactivation domain was inactive. The viral protein E1A also prevented the action of each of the three components in a manner that was independent of E1A's ability to block the histone acetyltransferase activity of CBP. Collectively, these results suggest that three different inputs (GME, GR, and coactivator) for perturbing the dose-response curve, and partial agonist activity, of GR-steroid complexes act by converging at a single step that involves a limiting factor prior to transcription initiation.
authors.affiliation: Steroid Hormones Section, NIDDK/Laboratory of Molecular and Cellular Biology, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.forename: N J
authors.initials: NJ
authors.lastname: Sarlis
authors.forename: S S
authors.initials: SS
authors.lastname: Simons
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism; D017500:Adenovirus E1A Proteins / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D050882:CREB-Binding Protein; D003907:Dexamethasone / Q000494:pharmacology; D005938:Glucocorticoids / Q000494:pharmacology; D051548:Histone Acetyltransferases; D007700:Kinetics; D009687:Nuclear Proteins / Q000378:metabolism; D051958:Nuclear Receptor Coactivator 2; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014444:Tyrosine Transaminase / Q000235:genetics*
chemical_list: D017500:Adenovirus E1A Proteins; D005938:Glucocorticoids; C498003:Ncoa2 protein, rat; D009687:Nuclear Proteins; D051958:Nuclear Receptor Coactivator 2; D011965:Receptors, Glucocorticoid; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D003907:Dexamethasone; D000123:Acetyltransferases; D050882:CREB-Binding Protein; C494365:Crebbp protein, rat; D051548:Histone Acetyltransferases; D014444:Tyrosine Transaminase