Binary developmental decisions and homeostatic regulation by steroids require negative transcriptional regulation to balance steroid-mediated stimulatory effects. Human glucocorticoid receptor mutants were used to identify regions important for trans-repression of the gene encoding the alpha subunit of chorionic gonadotropin. While the amino terminus is not critical, the DNA binding and ligand binding domains are required for efficient repression. However, the function of the carboxyl terminus can be substituted by a polypeptide from the human mineralocorticoid receptor or beta-galactosidase gene. The function of these fusion repressors supports the model that the human glucocorticoid receptor negatively regulates transcription via a steric hindrance mechanism. These results suggest a potentially general strategy for creation of sequence-specific transcriptional repressors.