Partial androgen insensitivity caused by an androgen receptor mutation at amino acid 907 (Gly-->Arg) that results in decreased ligand binding affinity and reduced androgen receptor messenger ribonucleic acid levels

• Published in: The journal of clinical endocrinology and metabolism Vol. 81; no. 1; pp. 236 - 243
• Main Authors: Choong, C S, Sturm, M J, Strophair, J A, McCulloch, R K, Tilley, W D, Leedman, P J, Hurley, D M
• Format: Journal Article
• Language: English
• Published: United States 01.01.1996
• Subjects: Adult; Androgens - metabolism; Animals; Base Sequence; Disorders of Sex Development - genetics; Female; Genetic Linkage; Humans; Ligands; Mice; Molecular Sequence Data; Point Mutation; Receptors, Androgen - genetics; Receptors, Androgen - metabolism; RNA, Messenger - analysis; Transcriptional Activation; X Chromosome
• ISSN: 0021-972X
• DOI: 10.1210/jc.81.1.236

Androgen insensitivity is an X-linked disorder of sexual differentiation resulting from mutations in the androgen receptor (AR) gene. In this paper, we report the clinical phenotype and molecular analysis of two siblings with severe partial androgen insensitivity due to a novel mutation in the ligand-binding domain of the AR gene. Binding studies using cultured genital skin fibroblasts demonstrated reduced AR affinity and binding capacity. Nucleotide sequence analysis of the AR gene of both siblings revealed a point mutation causing a glycine to arginine amino acid substitution at position 907 within a conserved region of the ligand-binding domain. A silent guanine to adenine substitution was also identified in the protein-coding region of exon 1. Using an expression vector in which the identified mutation was recreated by site-directed mutagenesis, the mutant receptor was found to have a reduced binding affinity (Kd = 3.06 nmol/L) for mibolerone compared with that of normal AR (Kd = 1.71 nmol/L) when expressed in COS-7 cells. In cotransfection experiments using CV-1 cells and a mouse mammary tumor virus-chloramphenicol acetyltransferase reporter system, the concentration of dihydrotestosterone required to induce half-maximal chloramphenicol acetyltransferase gene expression was 50-fold higher in cells transfected with the mutant AR complementary DNA than in cells transfected with normal AR complementary DNA. AR messenger ribonucleic acid levels in genital skin fibroblasts determined by both competitive PCR amplification and ribonuclease protection assay were decreased compared with normal values. Our studies demonstrate the importance of this region of the AR gene in normal AR function and AR gene expression.