title: A novel point mutation in the ligand-binding domain (LBD) of the human glucocorticoid receptor (hGR) causing generalized glucocorticoid resistance: the importance of the C terminus of hGR LBD in conferring transactivational activity.
abstract: Glucocorticoid resistance is a rare, familial or sporadic condition characterized by partial end-organ insensitivity to glucocorticoids. The clinical spectrum of the condition is broad, ranging from completely asymptomatic to severe hyperandrogenism and/or mineralocorticoid excess. The molecular basis of glucocorticoid resistance has been ascribed to mutations in the human glucocorticoid receptor-alpha (hGRalpha) gene, which impair one or more of the molecular mechanisms of GR action, thus altering tissue sensitivity to glucocorticoids. We identified a new case of generalized glucocorticoid resistance in a young woman who presented with a long-standing history of fatigue, anxiety, hyperandrogenism, and hypertension. The disease was caused by a novel, heterozygous mutation (T-->C) at nucleotide position 2318 (exon 9) of the hGRalpha gene, which resulted in substitution of leucine by proline at amino acid position 773 in the ligand-binding domain of the receptor. We systematically investigated the molecular mechanisms through which the natural hGRalphaL773P mutant impaired glucocorticoid signal transduction. Compared with the wild-type hGRalpha, hGRalphaL773P demonstrated a 2-fold reduction in the ability to transactivate the glucocorticoid-inducible mouse mammary tumor virus promoter, exerted a dominant negative effect on the wild-type receptor, had a 2.6-fold reduction in the affinity for ligand, showed delayed nuclear translocation (30 vs. 12 min), and, although it preserved its ability to bind to DNA, displayed an abnormal interaction with the GR-interacting protein 1 coactivator in vitro. We conclude that the carboxyl terminus of the ligand-binding domain of hGRalpha is extremely important in conferring transactivational activity by altering multiple functions of this composite transcription factor.
authors.affiliation: Department of Pediatric Endocrinology, Great Ormond Street Hospital for Children, 9th Floor, Southwood Building, Great Ormond Street, London, United Kingdom WC1N 3JH. charmane@mail.nih.gov
authors.forename: Evangelia
authors.initials: E
authors.lastname: Charmandari
authors.forename: Annaswamy
authors.initials: A
authors.lastname: Raji
authors.forename: Tomoshige
authors.initials: T
authors.lastname: Kino
authors.forename: Takamasa
authors.initials: T
authors.lastname: Ichijo
authors.forename: Anatoly
authors.initials: A
authors.lastname: Tiulpakov
authors.forename: Keith
authors.initials: K
authors.lastname: Zachman
authors.forename: George P
authors.initials: GP
authors.lastname: Chrousos
mesh_terms: D000328:Adult; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D003907:Dexamethasone / Q000493:pharmacokinetics / Q000494:pharmacology; D004351:Drug Resistance; D005260:Female; D017930:Genes, Reporter; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D017354:Point Mutation / Q000235:genetics*; D011965:Receptors, Glucocorticoid / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013936:Thymidine / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics*; D014162:Transfection
chemical_list: D017931:DNA Primers; D005938:Glucocorticoids; D008024:Ligands; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone; D013936:Thymidine