Functional characterization of the hGRαT556I causing Chrousos syndrome

Background Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor ( hGR ) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incidentaloma, who harboured a heterozygous point mutation in the hGR gene, which resulted in threonine (T) to isoleucine (I) substitution at amino acid position 556 in the ligand‐binding domain of the receptor. Objective To delineate the molecular mechanisms through which the mutant receptor hGRαT556I causes Chrousos syndrome. Design and Results Compared with the wild‐type receptor, the mutant receptor hGRαT556I demonstrated 50% reduction in its ability to transactivate glucocorticoid‐responsive genes and in the affinity for the ligand, 30% increase in the ability to transrepress the nuclear factor‐κB‐target genes and a 3,4‐fold delay in the cytoplasmic‐to‐nuclear translocation. The mutant receptor hGRαT556I did not exert a dominant negative effect upon the hGRα‐mediated transcriptional activity; it preserved its ability to bind to DNA and interacted with the glucocorticoid receptor‐interacting protein 1 coactivator mostly through its activation function‐1 domain. Structural biology studies revealed that the T556I mutation caused disruption of the hydrogen bond formed by the T556 with the =O group of P637 backbone, which resulted in a significant relocation of the P637‐bearing loop. This conformational alteration affected the local 3D arrangement of the receptor and hence the electrostatic surface of the region. Conclusions The hGRαT556I causes Chrousos syndrome by impairing multiple steps of the glucocorticoid signal transduction pathway.