Characterization of two novel mutations in the glucocorticoid receptor gene in patients with primary cortisol resistance.

摘要: OBJECTIVE Primary glucocorticoid resistance is characterized by decreased sensitivity to cortisol signalling. We have performed genetic analysis of the receptor (GR) gene in 12 unrelated patients with primary as defined a pathological dexamethasone suppression test. METHODS Exon specific polymerase chain reaction amplification GR and sequencing each exon was carried out. The two mutations were vitro terms driven reporter activity transient transfection assay ligand binding assay. Molecular modelling R477H mutant based on X-ray structure GR-DNA domain. RESULTS Two novel found: DNA-binding domain which first reported mutation that region human G679S domain. showed no transactivating capacity, whereas had reduced transactivation capacity compared wild-type (wt) GR. When tested for 50% affinity wt effect point deduced comparison between model mutant. has direct water mediated contact phosphate groups responsive element (GRE) whereas, model, GRE. located surface domain, at distance from steroid-binding site. A previously polymorphism, AAT AAC amino acid position 766, found four patients. CONCLUSIONS In clinical resistance, forms could be found. vivo these corresponds impaired function mutated assays. relevance conservative polymorphism insensitivity noted remains clarified.