title: Ala 586 Asp mutation in androgen receptor disrupts transactivation function without affecting androgen binding.
abstract: OBJECTIVE
  To understand the pathogenesis of the androgen insensitivity syndrome.
  
  
  METHODS
  Familial case study.
  
  
  METHODS
  Medical and Evolutionary Genetics Laboratory, Centre for Cellular and Molecular Biology, Hyderabad, India.
  
  
  METHODS
  Two affected sisters and other unaffected family members.
  
  
  METHODS
  The hormone levels were measured by RIA. Histology was done by standard protocols. DNA isolation and direct DNA sequencing was undertaken for mutation identification. Site-directed mutagenesis was used for incorporation of mutation in the androgen receptor clone. Functional assays were done using COS-1 cell cultures.
  
  
  METHODS
  Phenotype, hormone levels, DNA mutations, ligand binding, transactivation function of androgen-androgen receptor complex.
  
  
  RESULTS
  The patients exhibited a female phenotype despite the 46,XY chromosome complement. Both of the affected individuals had higher levels of T and LH. C1760A (coding DNA sequence reference) substitution (Ala 586 Asp) in the AR gene was observed in all of the affected individuals. The mutation did not result in a loss of ligand binding but instead in almost complete loss of transactivation function.
  
  
  CONCLUSIONS
  The Ala 586 Asp mutation resulted in a complete loss of transactivation function of the androgen-androgen receptor complex but did not affect ligand binding. In vitro assays confirmed the pathogenic nature of this mutation.
authors.affiliation: Centre for Cellular and Molecular Biology, Hyderabad, India. thangs@ccmb.res.in
authors.forename: Singh
authors.initials: S
authors.lastname: Rajender
authors.forename: Nalini J
authors.initials: NJ
authors.lastname: Gupta
authors.forename: Baidyanath
authors.initials: B
authors.lastname: Chakrabarty
authors.forename: Lalji
authors.initials: L
authors.lastname: Singh
authors.forename: Kumarasamy
authors.initials: K
authors.lastname: Thangaraj
mesh_terms: D000595:Amino Acid Sequence; D013734:Androgen-Insensitivity Syndrome / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000728:Androgens / Q000097:blood*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D005260:Female; D043373:Follicle Stimulating Hormone, Human / Q000097:blood; D020022:Genetic Predisposition to Disease; D006801:Humans; D007621:Karyotyping; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010375:Pedigree; D010641:Phenotype; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D013739:Testosterone / Q000097:blood; D015533:Transcriptional Activation; D014162:Transfection; D055815:Young Adult
chemical_list: C410798:AR protein, human; D000728:Androgens; D043373:Follicle Stimulating Hormone, Human; D011944:Receptors, Androgen; D013739:Testosterone; D007986:Luteinizing Hormone