Screening of EDA1 gene in X-linked anhidrotic ectodermal dysplasia using DHPLC:: Identification of 14 novel mutations in Italian patients
Screening of EDA1 gene in X-linked anhidrotic ectodermal dysplasia using DHPLC:: Identification of 14 novel mutations in Italian patients
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DOI：
10.1089/gte.2008.0020
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发表时间：
2008-09-01
期刊：
影响因子：
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通讯作者：
Novelli, Giuseppe
中科院分区：
文献类型：
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作者：
Conte, Chiara;Gambardella, Stefano;Novelli, Giuseppe
编码外胚层发育异常蛋白的EDA1基因发生突变，导致x连锁无汗性外胚层发育不良。本研究分析了23例意大利无汗性外胚层发育不良患者的EDA1基因突变。我们通过变性高效液相色谱建立了快速方案，随后进行测序，允许表征18个突变，14个新突变和4个复发突变：8个错义突变（p.L51Q、p.H54R、p.R156H两次、p.C332F、p.D316H、p.T378M和p.A349T）、3个框内缺失（p.G82_P84del、p.A179_P191del和p.L354del）、1个总缺失（p.G168_G265del，通过直接测序和PCR鉴定）、4个剪接改变（C .949- 13t > C、C .741 + 1G/T、C . 793 + 4A> T和C . 924 + 1G/T）、1个无义突变（p.Y3X）和1个同义突变（C . 741g > A）。此外，对三个错义突变的结构分析表明，蛋白质静电表面的改变（p.D316N），单体间相互作用的破坏（p.A349T）和单体结构的不稳定（p.T378M）可能不可逆地使EDAA1的结合特性失效。我们的数据证实并扩展了EDA1突变的大谱，为检测EDA患者的EDA1突变提供了一种快速有效的分子方案。
Mutations within EDA1 gene, which encodes for the ectodysplasin, cause X-linked anhidrotic ectodermal dysplasia. In this study, 23 Italian patients with anhidrotic ectodermal dysplasia were analyzed for mutations in EDA1 gene. We set up a rapid protocol through denaturing high-performance liquid chromatography, followed by sequencing, that allowed the characterization of 18 mutations, 14 novel and 4 recurrent: 8 missense mutations (p.L51Q, p.H54R, p.R156H twice, p.C332F, p.D316H, p.T378M, and p.A349T), 3 in-frame deletions (p.G82_P84del, p.A179_P191del, and p.L354del), 1 gross deletion (p.G168_G265del, identified through direct sequencing and PCR), 4 altered splicing (c.949-13T> C, c. 741 + 1G/T, c. 793 + 4A> T, and c. 924 + 1G/T), 1 nonsense (p.Y3X), and 1 synonymous mutation (c.741G> A). Moreover, structural analysis of three missense mutations shows that alteration of the electrostatic surface of the protein (p.D316N), the break of intermonomer interactions (p.A349T) and destabilization of the single monomer structure (p.T378M), may irreversibly invalidate the EDAA1 binding properties. Our data confirm and extend the large spectrum of EDA1 mutations and provide a rapid and efficient molecular protocol for testing EDA1 mutations in EDA patients.