Massively Parallel Functional Analysis of BRCA1 RING Domain Variants
Massively Parallel Functional Analysis of BRCA1 RING Domain Variants
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DOI：
10.1534/genetics.115.175802
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发表时间：
2015-06-01
期刊：
影响因子：
3.3
通讯作者：
Fields, Stanley
中科院分区：
文献类型：
--
作者：
Starita, Lea M.;Young, David L.;Fields, Stanley
解释不确定意义变异（VUS）是医学遗传学的核心挑战。一种方法是实验测量VUS的功能后果，但迄今为止，这种方法是事后的和低通量的。在这里，我们使用大量平行分析来测量BRCA1的环结构域中近2000个错义替换对其E3泛素连接酶活性及其与BARD1环结构域结合的影响。从结果得分中，我们生成了一个模型来预测全长BRCA1变体支持同源性定向DNA修复的能力，BRCA1在肿瘤抑制中的重要作用，并表明它优于广泛使用的生物效应预测算法。我们设想，大规模平行功能分析可能有助于对临床测序中观察到的变异进行前瞻性解释。
Interpreting variants of uncertain significance (VUS) is a central challenge in medical genetics. One approach is to experimentally measure the functional consequences of VUS, but to date this approach has been post hoc and low throughput. Here we use massively parallel assays to measure the effects of nearly 2000 missense substitutions in the RING domain of BRCA1 on its E3 ubiquitin ligase activity and its binding to the BARD1 RING domain. From the resulting scores, we generate a model to predict the capacities of full-length BRCA1 variants to support homology-directed DNA repair, the essential role of BRCA1 in tumor suppression, and show that it outperforms widely used biological-effect prediction algorithms. We envision that massively parallel functional assays may facilitate the prospective interpretation of variants observed in clinical sequencing.