title: Functional effects of DAX-1 mutations identified in patients with X-linked adrenal hypoplasia congenita.
abstract: X-linked adrenal hypoplasia congenita with hypogonadotropic hypogonadism and adrenal insufficiency is a rare disorder caused by mutations of DAX-1. In this study, we investigated the functional defects of DAX-1 caused by mutations identified in 3 unrelated Korean patients with adrenal hypoplasia congenita. The DAX-1 gene was directly sequenced using genomic DNA isolated from peripheral blood leukocytes. The functional defects of DAX-1 caused by mutations were evaluated using an in vitro promoter assay. After mutagenesis of DAX-1 complementary DNA in the pcDNA3.1 vector, steroidogenic factor 1 and the promoter region of steroidogenic acute regulatory protein (StAR) genes in pGL4.10[luc2] were transiently cotransfected into human embryonic kidney 293 cells, followed by luminometry measurements of the luciferase activity of StAR. Mutation analysis of 3 patients revealed p.L386delfsX2, p.W105X, and p.Q252X mutations of the DAX-1 gene. The mutant DAX-1 proteins showed lower repressive activity on the StAR gene promoter when compared with normal DAX-1. Nonsense and frameshift mutations of the DAX-1 gene partially eliminated the ability of DAX-1 to repress the transcription of StAR in an in vitro assay.
authors.affiliation: Division of Pediatric Endocrinology and Metabolism, Department of Pediatrics, Asan Medical Center Children's Hospital, University of Ulsan College of Medicine, Seoul, South Korea.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Choi
authors.forename: Jung-Young
authors.initials: JY
authors.lastname: Park
authors.forename: Gu-Hwan
authors.initials: GH
authors.lastname: Kim
authors.forename: Hye Young
authors.initials: HY
authors.lastname: Jin
authors.forename: Beom Hee
authors.initials: BH
authors.lastname: Lee
authors.forename: Jae Hyun
authors.initials: JH
authors.lastname: Kim
authors.forename: Choong Ho
authors.initials: CH
authors.lastname: Shin
authors.forename: Sei Won
authors.initials: SW
authors.lastname: Yang
authors.forename: Han-Wook
authors.initials: HW
authors.lastname: Yoo
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000235:genetics*; D000309:Adrenal Insufficiency; D001483:Base Sequence; D002478:Cells, Cultured; D002648:Child; D057137:DAX-1 Orphan Nuclear Receptor / Q000235:genetics* / Q000502:physiology*; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation / Q000235:genetics; D040181:Genetic Diseases, X-Linked / Q000235:genetics*; D006801:Humans; D000075262:Hypoadrenocorticism, Familial; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology; D010375:Pedigree; D010750:Phosphoproteins / Q000235:genetics; D014162:Transfection; D055815:Young Adult; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D057137:DAX-1 Orphan Nuclear Receptor; C539965:NR0B1 protein, human; D010750:Phosphoproteins; D000098682:Steroidogenic Acute Regulatory Protein