title: Massively Parallel Functional Analysis of BRCA1 RING Domain Variants.
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.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: Lea M
authors.initials: LM
authors.lastname: Starita
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: David L
authors.initials: DL
authors.lastname: Young
authors.affiliation: Department of Biomedical Informatics and The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.
authors.forename: Muhtadi
authors.initials: M
authors.lastname: Islam
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: Jacob O
authors.initials: JO
authors.lastname: Kitzman
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: Justin
authors.initials: J
authors.lastname: Gullingsrud
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Hause
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Fowler
authors.affiliation: 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.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Parvin
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195 fields@uw.edu shendure@uw.edu Jeffrey.Parvin@osumc.edu.
authors.forename: Jay
authors.initials: J
authors.lastname: Shendure
authors.affiliation: 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.
authors.forename: Stanley
authors.initials: S
authors.lastname: Fields
mesh_terms: D019313:BRCA1 Protein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004260:DNA Repair; D014644:Genetic Variation; D023281:Genomics / Q000379:methods; D061325:Hereditary Breast and Ovarian Cancer Syndrome / Q000235:genetics / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009154:Mutation; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D055503:Protein Multimerization; D044767:Ubiquitin-Protein Ligases
chemical_list: D019313:BRCA1 Protein; D044767:Ubiquitin-Protein Ligases
keywords: BRCA1; deep mutational scanning; human genetic variation; protein function; variants of uncertain significance
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