Vilchis Felipe, Ramos Luis, Kofman-Alfaro Susana, Zenteno Juan Carlos, Méndez Juan Pablo, Chávez Bertha
Department of Reproductive Biology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga 15, Tlalpan C.P. 14000, México D.F., México.
J Hum Genet. 2003;48(7):346-51. doi: 10.1007/s10038-003-0036-0. Epub 2003 Jun 7.
Androgen insensitivy syndrome (AIS) is the most frequent cause of male pseudohermaphroditism resulting from target-organ resistance to androgen action. Individuals bearing the complete form of the disease (CAIS) present a female phenotype and a lack of pubic and axillary hair. In the present study, four 46,XY patients born in two generations from a kindred with a history of AIS were examined for genetic abnormalities in the androgen receptor gene (AR). All eight exons encoding the AR protein were individually amplified from genomic DNA followed by a mutation screening with single-strand conformation polymorphism analysis. Sequencing of the mutant AR revealed a novel insertion/deletion mutation in exon 5. A deletion of 7 bp is replaced by an insertion of 11 nucleotides, which represents a duplication of the adjacent downstream sequence. The mutation g.2640_2646delAGGATGC/2652_2662insTTCGCCCCTGA, results in a frameshift that introduces a premature termination signal TGA, nine codons downstream. Such a rearrangement predicts a truncation of the AR, thereby deleting a large portion of the ligand-binding domain (amino acid position 768-919). Furthermore, although this mutation breaks the translational reading frame starting from codon 760, examination of the complementary DNA suggested that it does not disturb mRNA splicing. These changes have been found in all the patients and appear to account for the observed absence of detectable androgen binding to the AR in cultured fibroblasts and for the CAIS phenotype in the kindred. This disorder represents the first insertion/deletion mutation of the AR that probably arose by a slipped-strand mispairing mechanism.
雄激素不敏感综合征(AIS)是男性假两性畸形最常见的病因,由靶器官对雄激素作用的抵抗所致。患有完全型疾病(CAIS)的个体表现为女性表型,且无阴毛和腋毛。在本研究中,对来自一个有AIS病史家系的两代出生的4名46,XY患者进行了雄激素受体基因(AR)的基因异常检测。从基因组DNA中分别扩增出编码AR蛋白的所有8个外显子,然后用单链构象多态性分析进行突变筛查。对突变的AR进行测序,发现外显子5中有一个新的插入/缺失突变。一个7 bp的缺失被11个核苷酸的插入所取代,这代表了相邻下游序列的重复。突变g.2640_2646delAGGATGC/2652_2662insTTCGCCCCTGA导致移码,在下游9个密码子处引入了一个过早的终止信号TGA。这种重排预测AR会被截断,从而删除了大部分配体结合域(氨基酸位置768 - 919)。此外,虽然这个突变破坏了从密码子760开始的翻译阅读框,但对互补DNA的检测表明它不会干扰mRNA剪接。在所有患者中都发现了这些变化,这似乎可以解释在培养的成纤维细胞中观察到的雄激素与AR结合检测不到的现象,以及该家系中的CAIS表型。这种疾病代表了AR的首次插入/缺失突变,可能是由滑链错配机制引起的。