Steroid hormones are small lipophilic molecules that control a wide range of responses in both the developing and adult organism. The actions of these molecules are mediated by soluble receptor proteins that function as hormone-activated transcription factors. The first steroid receptors were expressed in the yeast Saccharomyces cerevisiae over 10 years ago, and to date virtually all the classical steroid receptors, together with a number of non-steroid members of the nuclear receptor superfamily, have been expressed in yeast. The ability to reconstitute steroid receptor signalling in yeast cells by co-expression of the receptor protein and a reporter gene driven by the appropriate hormone response element has presented researchers with a powerful model system for investigating receptor action. In this review, the use of yeast-based steroid receptor transactivation assays to investigate the roles of molecular chaperones, the mechanisms of DNA binding and gene activation, and the functional properties of hormone mimics will be discussed.