Format

Send to

Choose Destination
Genetics. 2015 Jun;200(2):413-22. doi: 10.1534/genetics.115.175802. Epub 2015 Mar 30.

Massively Parallel Functional Analysis of BRCA1 RING Domain Variants.

Author information

1
Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
2
Department of Biomedical Informatics and The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.
3
Department of Biomedical Informatics and The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210 fields@uw.edu shendure@uw.edu Jeffrey.Parvin@osumc.edu.
4
Department of Genome Sciences, University of Washington, Seattle, Washington 98195 fields@uw.edu shendure@uw.edu Jeffrey.Parvin@osumc.edu.
5
Department of Genome Sciences, University of Washington, Seattle, Washington 98195 Department of Medicine, University of Washington, Seattle, Washington 98195 Howard Hughes Medical Institute, University of Washington, Seattle, Washington 98195 fields@uw.edu shendure@uw.edu Jeffrey.Parvin@osumc.edu.

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.

KEYWORDS:

BRCA1; deep mutational scanning; human genetic variation; protein function; variants of uncertain significance

PMID:
25823446
PMCID:
PMC4492368
DOI:
10.1534/genetics.115.175802
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center