title: Promoter-dependent activity on androgen receptor N-terminal domain mutations in androgen insensitivity syndrome.
abstract: Androgen receptor (AR) mutations are associated with androgen insensitivity syndrome (AIS). Missense mutations identified in the AR-N-terminal domain (AR-NTD) are rare, and clinical phenotypes are typically mild. We investigated 7 missense mutations and 2 insertion/deletions located in the AR-NTD. This study aimed to elucidate the pathogenic role of AR-NTD mutants in AIS and to use this knowledge to further define AR-NTD function. AR-NTD mutations (Q120E, A159T, G216R, N235K, G248V, L272F, and P380R) were introduced into AR-expression plasmids. Stably expressing cell lines were established for del57L and ins58L. Transactivation was measured using luciferase reporter constructs under the control of GRE and Pem promoters. Intrinsic fluorescence spectroscopy and partial proteolysis studies were performed for mutations which showed reduced activities by using a purified AR-AF1 protein. Pem-luciferase reporter activation was reduced for A159T, N235K, and G248V but not the GRE-luciferase reporter. Protein structure analysis detected no significant change in the AR-AF1 region for these mutations. Reduced cellular expression and transactivation activity were observed for ins58L. The mutations Q120E, G216R, L272F, P380R, and del57L showed small or no detectable changes in function. Thus, clinical and experimental analyses have identified novel AR-signalling defects associated with mutations in the structurally disordered AR-NTD domain in patients with AIS.
authors.affiliation: Department of Paediatrics, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Rieko
authors.initials: R
authors.lastname: Tadokoro-Cuccaro
authors.forename: John
authors.initials: J
authors.lastname: Davies
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Mongan
authors.forename: Trevor
authors.initials: T
authors.lastname: Bunch
authors.forename: Rosalind S
authors.initials: RS
authors.lastname: Brown
authors.forename: Laura
authors.initials: L
authors.lastname: Audi
authors.forename: Kate
authors.initials: K
authors.lastname: Watt
authors.forename: Iain J
authors.initials: IJ
authors.lastname: McEwan
authors.forename: Ieuan A
authors.initials: IA
authors.lastname: Hughes
mesh_terms: D013734:Androgen-Insensitivity Syndrome / Q000235:genetics*; D013196:Dihydrotestosterone / Q000494:pharmacology; D005260:Female; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D059748:Proteolysis / Q000187:drug effects; D011944:Receptors, Androgen / Q000737:chemistry* / Q000235:genetics*; D013050:Spectrometry, Fluorescence; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics
chemical_list: C410798:AR protein, human; D050505:Mutant Proteins; D011944:Receptors, Androgen; D013196:Dihydrotestosterone