Molecular actions of estrogen agonists and antagonists and their interaction with the human estrogen receptor

Studies were undertaken to examine the interaction of estrogen agonists and antagonists with the human estrogen receptor (ER) and to examine the consequences of this interaction on the DNA binding and transactivating ability of the receptor. The functional role of cysteines in the hormone binding domain (HBD) of the receptor was examined by in vitro mutagenesis, followed by analysis of the hormone binding, covalent labeling and transactivating ability of the mutant receptors. Mutagenesis revealed that Cys530 is the preferential site of labeling by covalent affinity labels, however, in the absence of a cysteine residue at postion 530, Cys 381 is labeled, and Cys381 lies near the ligand binding pocket. Receptors bearing single or multiple cysteine to alanine substitutions were not altered in their response to reversibly binding ligands. However, the effectiveness of covalently attaching ligands were altered in receptors containing an alanine at position 530, indicating an important role of Cys530 in the function of these ligands. One receptor mutant (C447A), displayed an estradiol dose response shift in transactivation assays of about 30-50 fold. Analysis of the C447A mutant revealed that this receptor is a temperature-sensitive mutant whose DNA binding is ligand-dependent at elevated temperatures.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Molecular and Integrative Physiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Reese, Joseph Charles, Jr.

Contributors dc:contributor

• Katzenellenbogen, Benita S.

• Biology, Animal Physiology

Statement dc:rights

• Copyright 1992 Reese, Joseph Charles, Jr

Language dc:language

eng

Identifier

AAI9236575
(UMI)AAI9236575

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/20261