Rapidly screening variants of uncertain significance in the MAP3K1 gene for phenotypic effects
Rapidly screening variants of uncertain significance in the MAP3K1 gene for phenotypic effects
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DOI：
10.1111/j.1399-0004.2011.01834.x
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发表时间：
2012-03-01
影响因子：
3.5
通讯作者：
Ostrer, H.
中科院分区：
文献类型：
--
作者：
Loke, J.;Ostrer, H.
在诊断或研究基础上对候选基因或整个外显子组进行DNA测序将产生大量意义不确定的变体，其潜在的表型作用不能通过预测程序、SNP数据库或常规遗传分析容易地证明。许多变体可通过影响数量、翻译后修饰或蛋白质相互作用而在编码蛋白质中产生表型变化。在这里，我们建立了免疫沉淀后流式细胞术方法的应用，以显示已知的蛋白质相互作用在46，XY性腺发育不全患者的B-淋巴母细胞样细胞中发生了改变，该患者的性腺发育不全是由MAP 3 K1基因突变引起的。该方法可以容易地扩展以同时测试来自可用组织的许多变体的多种相互作用，以及量化变体对蛋白质积累和翻译后修饰的影响，从而提供用于筛选对于表型效应具有不确定意义的变体的有效手段。
DNA sequencing of candidate genes or whole exomes on a diagnostic or investigational basis will yield a plethora of variants of uncertain significance whose potential phenotypic roles cannot be readily demonstrated by prediction programs, SNP databases nor conventional genetic analysis. Many variants may produce phenotypic changes in the encoded proteins by affecting the quantity, post- translational modification or protein interactions. Here, we establish the application of the method of flow cytometry following immunoprecipitation to show that known protein interactions are altered in the B- lymphoblastoid cells of patients with 46, XY gonadal dysgenesis arising from mutations in the MAP3K1 gene. This method can be scaled readily to test multiple interactions for many variants simultaneously from available tissues as well as quantify the effects of variants on protein accumulation and post- translational modification, thus providing an efficient means for screening variants of uncertain significance for phenotypic effects.