A genetically modified glucocorticoid receptor which is transactivation deficient is used to identify cofactors which will be useful to provide inflammation-inhibiting and immunosuppressive treatment

摘要

A genetically modified glucocorticoid receptor, which has transrepression protein-protein reciprocal effects and is transactivation deficient, is new. Independent claims are also included for: (1) a gene construct comprising at least a nucleic acid encoding the claimed receptor, operably linked with regulatory sequences of a reporter gene, preferably a DNA-binding domain for a reporter gene; (2) identifying a gene encoding a cofactor involved in glucocorticoid receptor modulation of at least another transcription factor (cross talk) comprising: (a) using the above construct with an expression bank of a eukaryotic cell expressed in a yeast two hybrid system; (b) detecting a specific protein-protein complex or the receptor and a cofactor through growth in a selective medium for the reporter; and (c) isolating and characterizing the nucleic acid encoding the cofactor in the cDNA clone; (3) a cofactor with transrepression specific for the glucocorticoid receptor which in a protein-protein interaction achieves a reciprocal effect, whereby within a downstream segment the N-terminal AF-1 and the DNA-binding domain of the receptors are bound; (4) identifying an agent which affects the reciprocal effect of the glucocorticoid receptor with other transcription factors and/or cofactors, where by the claimed receptor or construct is contacted with a potential agent and modulation of the interaction of the protein partner is measured by expression of the reporter gene or detecting protein-protein complex binding; (5) an agent for modulating interaction of the glucocorticoid receptor with a cofactor which binds either at the binding site of a physiological hormone or at a separate binding place; and (6) a compound with an inflammation-inhibiting or immunosuppressive effect comprising the above agent.