Massively Parallel Functional Analysis of BRCA1 RING Domain Variants

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • BRCA1 Protein / chemistry
  • BRCA1 Protein / genetics*
  • BRCA1 Protein / metabolism*
  • DNA Repair
  • Genetic Variation*
  • Genomics / methods
  • Hereditary Breast and Ovarian Cancer Syndrome / genetics
  • Hereditary Breast and Ovarian Cancer Syndrome / metabolism
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Mutation
  • Protein Interaction Domains and Motifs* / genetics
  • Protein Multimerization
  • Ubiquitin-Protein Ligases

Substances

  • BRCA1 Protein
  • Ubiquitin-Protein Ligases