Massively Parallel Functional Analysis of BRCA1 RING Domain Variants

Overview

Journal Genetics

Publisher Oxford University Press

Specialty Genetics

Date 2015 Apr 1

PMID 25823446

Citations 166

Authors

Lea M Starita

David L Young

Muhtadi Islam

Jacob O Kitzman

Justin Gullingsrud

Ronald J Hause

Douglas M Fowler

Jeffrey D Parvin

Jay Shendure

Stanley Fields

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

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Higher throughput assays for understanding the pathogenicity of variants of unknown significance (VUS) in the RPE65 gene.

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