title: Complete androgen insensitivity caused by a splice donor site mutation in intron 2 of the human androgen receptor gene resulting in an exon 2-lacking transcript with premature stop-codon and reduced expression.
abstract: Various mutations within the human androgen receptor gene have been documented to cause defective sexual differentiation in karyotypic male individuals. In this study, we report a previously undescribed point mutation at the donor splice-site of the second intron of the androgen receptor gene in a patient with a completely female phenotype. The sequence alteration was detected by single-strand-conformation-analysis-PCR and genomic sequencing. Applying competitive reverse transcribed PCR, cDNA sequencing and Western blotting, we could demonstrate considerable aberrations of structure and concentration of the transcript and its translation product in the patient's fibroblasts from the genital region. (1) In the transcript, exon 1 and 3 are directly linked to each other, the complete second exon is skipped. The mRNA predictively suffers a codon frame-shift in exon 3 associated with a premature termination between codons 598 and 599, leading to a truncated androgen receptor protein lacking any in vivo function. (2) Steady-state concentration levels of transcript and protein are abnormally low. Our observations highlight the influence of exon-flanking intron sequences on proper expression and function of gene products.
authors.affiliation: Department of Paediatrics, Medical University of Lübeck, Germany.
authors.forename: O J
authors.initials: OJ
authors.lastname: Hellwinkel
authors.forename: K
authors.initials: K
authors.lastname: Bull
authors.forename: P M
authors.initials: PM
authors.lastname: Holterhus
authors.forename: N
authors.initials: N
authors.lastname: Homburg
authors.forename: D
authors.initials: D
authors.lastname: Struve
authors.forename: O
authors.initials: O
authors.lastname: Hiort
mesh_terms: D000595:Amino Acid Sequence; D013734:Androgen-Insensitivity Syndrome / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D007438:Introns; D008297:Male; D008969:Molecular Sequence Data; D010641:Phenotype; D017354:Point Mutation; D018807:Polymorphism, Single-Stranded Conformational; D014176:Protein Biosynthesis; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012733:Sex Differentiation / Q000235:genetics
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D011944:Receptors, Androgen