title: Natural glucocorticoid receptor mutants causing generalized glucocorticoid resistance: molecular genotype, genetic transmission, and clinical phenotype.
abstract: Glucocorticoid resistance is a rare, familial, or sporadic condition characterized by partial end-organ insensitivity to glucocorticoids. The clinical spectrum of the condition ranges from completely asymptomatic to severe hyperandrogenism, fatigue, and/or mineralocorticoid excess. The molecular basis of glucocorticoid resistance in several families and sporadic cases has been ascribed to mutations in the human glucocorticoid receptor-alpha (hGRalpha) gene, which impair the ability of the receptor to transduce the glucocorticoid signal. We systematically investigated the molecular mechanisms through which natural, ligand-binding domain hGRalpha mutants, including hGRalphaI559N, hGRalphaV571A, hGRalphaD641V, hGRalphaV729I, and hGRalphaI747M, produce a defective signal and determined whether their differential effects on hGRalpha function might account for the type of genetic transmission of the disorder and the variable clinical phenotype of the affected subjects. Our findings suggest that all five mutant receptors studied have ligand-binding domains with decreased intrinsic transcriptional activity. Unlike hGRalphaI559N and I747M previously shown to exert a dominant negative effect upon the transcriptional activity of hGRalpha, hGRalphaV571A, D641V, and V729I do not have such an effect. All five mutants studied demonstrate varying degrees of decreased affinity for the ligand in a standard dexamethasone binding assay, but preserve their ability to bind DNA. The nondominant negative mutants, hGRalphaV571A, D641V, and V729I, show delayed translocation into the nucleus after exposure to ligand. Finally, hGRalphaI559N, V571A, D641V, and V729I display an abnormal interaction with the glucocorticoid receptor-interacting protein-1 coactivator in vitro, as this was previously shown also for hGRalphaI747M. We conclude that each of the above hGRalpha mutations imparts different functional defects upon the glucocorticoid signal transduction pathway, which explains the autosomal recessive or dominant transmission of the disorder, but might only explain in part its variable clinical phenotype.
authors.affiliation: Pediatric and Reproductive Endocrinology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Evangelia
authors.initials: E
authors.lastname: Charmandari
authors.forename: Tomoshige
authors.initials: T
authors.lastname: Kino
authors.forename: Emmanuil
authors.initials: E
authors.lastname: Souvatzoglou
authors.forename: Alessandra
authors.initials: A
authors.lastname: Vottero
authors.forename: Nisan
authors.initials: N
authors.lastname: Bhattacharyya
authors.forename: George P
authors.initials: GP
authors.lastname: Chrousos
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics; D005799:Genes, Dominant; D005838:Genotype; D005938:Glucocorticoids / Q000378:metabolism / Q000494:pharmacology*; D049452:Green Fluorescent Proteins; D006801:Humans; D008024:Ligands; D008164:Luminescent Proteins / Q000235:genetics; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010641:Phenotype; D020033:Protein Isoforms / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000502:physiology; D018091:Receptors, AMPA / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C491310:GRIP1 protein, human; D005938:Glucocorticoids; D008024:Ligands; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D018091:Receptors, AMPA; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins; C411470:glucocorticoid receptor alpha; D049452:Green Fluorescent Proteins; D003907:Dexamethasone