Genetic Approaches to Mammalian Nuclear Receptor Function in Yeast

Abstract Mammalian nuclear receptor function can be faithfully reconstituted in yeast, enabling a wide variety of genetic approaches to be taken toward defining the mechanisms of signal transduction and transcriptional regulation. This report describes vectors for the expression of mammalian receptors in yeast, reporter genes, yeast host strains, and simple assays that monitor receptor transcriptional activity. Methods for the generation of receptors with distinct defects in particular functions, such as DNA or hormone binding, that couple random mutagenesis with phenotypic screens are outlined as well. In addition, strategies for the identification of nonreceptor components whose gene products may act on receptors are discussed. The experimental advantages of yeast invite a detailed genetic analysis of mammalian nuclear receptor functions-hormone and DNA binding, nuclear localization, and interaction with nonreceptor factors-and should illuminate further the mechanisms of signal transduction and transcriptional regulation by this important class of regulatory molecules.