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.
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.
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DOI：
10.1210/mend.7.7.8413310
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发表时间：
1993-07
影响因子：
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通讯作者：
I. Mowszowicz;H. Lee;H. T. Chen;C. Mestayer;M. Portois;S. Cabrol;P. Mauvais‐Jarvis;C. Chang
中科院分区：
文献类型：
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作者：
I. Mowszowicz;H. Lee;H. T. Chen;C. Mestayer;M. Portois;S. Cabrol;P. Mauvais‐Jarvis;C. Chang
我们分析了两名受体阳性完全雄激素不敏感（Rec（+）-CAI）同胞（患者9006和9030）的人雄激素受体（AR）基因的互补DNA和基因组DNA的核苷酸序列。北方分析显示AR mRNA大小正常。然而，其表达在两个患者中相对减少。与正常雄激素结合能力（患者9006和9030分别为496和552 fmol/mg DNA）一致，但在生殖器皮肤成纤维细胞中测量的DNA结合能力降低（168 fmol/mg DNA），在AR的N末端和配体结合结构域均未发现突变。然而，在这两种情况下，在DNA结合结构域的第二锌指的核苷酸2372处的AR cDNA的单碱基取代（G->A）被发现。这导致高度保守的精氨酸残基（氨基酸614）被组氨酸取代。当突变的受体质粒与报告基因氯霉素乙酰转移酶基因一起共转染到PC-3细胞中时，氯霉素乙酰转移酶活性不被5 α-二氢睾酮治疗诱导，证实突变使AR无功能，因此可以对这些患者的临床特征负责。这些结果突出了精氨酸-614在蛋白质-DNA相互作用中AR的DNA结合结构域的第二锌指中的重要性。
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.