title: A point mutation in the second zinc finger of the DNA-binding domain of the androgen receptor gene causes complete androgen insensitivity in two siblings with receptor-positive androgen resistance.
abstract: We have analyzed the nucleotide sequence of complementary and genomic DNAs of the human androgen receptor (AR) gene in two siblings (patients 9006 and 9030) with receptor-positive complete androgen insensitivity (Rec(+)-CAI). Northern analysis indicated that mRNA of the AR was normal in size. However, its expression was relatively reduced in both patients. Consistent with the normal androgen-binding capacity (496 and 552 fmol/mg DNA for patients 9006 and 9030, respectively) but decreased DNA-binding ability (168 fmol/mg DNA) measured in genital skin fibroblasts, no mutation was found in both N-terminal and ligand-binding domains of the AR. However, a single base substitution (G-->A) was found in the second zinc finger of the DNA-binding domain at nucleotide 2372 of the AR cDNA in both cases. This resulted in the replacement of a highly conserved arginine residue (amino acid 614) by a histidine. When the mutated receptor plasmid was cotransfected into PC-3 cells together with the reporter chloramphenicol acetyltransferase gene, chloramphenicol acetyltransferase activity was not induced by 5 alpha-dihydrotestosterone treatment, confirming that the mutation renders the AR nonfunctional and can, therefore, be held responsible for the clinical features in these patients. These results highlight the importance of Arginine-614 in the second zinc finger of the DNA-binding domain of the AR in the protein-DNA interaction.
authors.affiliation: Biochemistry Laboratory B, Hospital Necker, Paris, France.
authors.forename: I
authors.initials: I
authors.lastname: Mowszowicz
authors.forename: H J
authors.initials: HJ
authors.lastname: Lee
authors.forename: H T
authors.initials: HT
authors.lastname: Chen
authors.forename: C
authors.initials: C
authors.lastname: Mestayer
authors.forename: M C
authors.initials: MC
authors.lastname: Portois
authors.forename: S
authors.initials: S
authors.lastname: Cabrol
authors.forename: P
authors.initials: P
authors.lastname: Mauvais-Jarvis
authors.forename: C
authors.initials: C
authors.lastname: Chang
mesh_terms: D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000032:analysis; D000293:Adolescent; D000595:Amino Acid Sequence; D000728:Androgens / Q000378:metabolism / Q000494:pharmacology*; D001120:Arginine / Q000032:analysis; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D004351:Drug Resistance; D005091:Exons; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D005784:Gene Amplification; D006639:Histidine / Q000032:analysis; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D010957:Plasmids; D017354:Point Mutation / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D000728:Androgens; D012333:RNA, Messenger; D011944:Receptors, Androgen; D006639:Histidine; D004247:DNA; D001120:Arginine; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase; D015500:Chloramphenicol O-Acetyltransferase