当前位置： SCI文献检索 > CLINICAL ENDOCRINOLOGY期刊下所有文献 > Characterization of two novel mutations in the glucocorticoid receptor gene in patients with primary cortisol resistance.

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

Abstract：

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

journal_name

Clin Endocrinol (Oxf)

journal_title

Clinical endocrinology

authors

Ruiz M,Lind U,Gåfvels M,Eggertsen G,Carlstedt-Duke J,Nilsson L,Holtmann M,Stierna P,Wikström AC,Werner S

doi

10.1046/j.1365-2265.2001.01323.x

keywords:

subject

Has Abstract

pub_date

2001-09-01 00:00:00

pages

363-71

issue

eissn

0300-0664

issn

1365-2265

pii

1323

journal_volume

pub_type

杂志文章

在线工具

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