Yeast-based reporter assays for the functional characterization of cochaperone interactions with steroid hormone receptors.
Steroid hormone receptor-mediated reporter assays in the budding yeast Saccharomyces cerevisiae have been an invaluable tool for the identification and functional characterization of steroid hormone receptor-associated chaperones and cochaperones. This chapter describes a hormone-inducible androgen receptor-mediated beta-galactosidase reporter assay in yeast. In addition, the immunophilin FKBP52 is used as a specific example of a receptor-associated cochaperone that acts as a positive regulator of receptor function. With the right combination of receptor and cochaperone expression plasmids, reporter plasmid, and ligand, the assay protocol described here could be used to functionally characterize a wide variety of nuclear receptor-cochaperone interactions. In addition to the functional characterization of receptor regulatory proteins, a modified version of this assay is currently being used to screen compound libraries for selective FKBP52 inhibitors that represent attractive therapeutic candidates for the treatment of steroid hormone receptor-associated diseases.
Balsiger
Heather A
HA
Department of Biological Sciences, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, USA.
Cox
Marc B
MB
D000818:Animals; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D017930:Genes, Reporter; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D018832:Molecular Chaperones; D011944:Receptors, Androgen; D011987:Receptors, Steroid; D001616:beta-Galactosidase; D022021:Tacrolimus Binding Proteins; C403485:tacrolimus binding protein 4