A mechanism for the suppression of homologous recombination in G1 cells

Nature. 2015 Dec 17;528(7582):422-6. doi: 10.1038/nature16142. Epub 2015 Dec 9.

Abstract

DNA repair by homologous recombination is highly suppressed in G1 cells to ensure that mitotic recombination occurs solely between sister chromatids. Although many homologous recombination factors are cell-cycle regulated, the identity of the events that are both necessary and sufficient to suppress recombination in G1 cells is unknown. Here we report that the cell cycle controls the interaction of BRCA1 with PALB2-BRCA2 to constrain BRCA2 function to the S/G2 phases in human cells. We found that the BRCA1-interaction site on PALB2 is targeted by an E3 ubiquitin ligase composed of KEAP1, a PALB2-interacting protein, in complex with cullin-3 (CUL3)-RBX1 (ref. 6). PALB2 ubiquitylation suppresses its interaction with BRCA1 and is counteracted by the deubiquitylase USP11, which is itself under cell cycle control. Restoration of the BRCA1-PALB2 interaction combined with the activation of DNA-end resection is sufficient to induce homologous recombination in G1, as measured by RAD51 recruitment, unscheduled DNA synthesis and a CRISPR-Cas9-based gene-targeting assay. We conclude that the mechanism prohibiting homologous recombination in G1 minimally consists of the suppression of DNA-end resection coupled with a multi-step block of the recruitment of BRCA2 to DNA damage sites that involves the inhibition of BRCA1-PALB2-BRCA2 complex assembly. We speculate that the ability to induce homologous recombination in G1 cells with defined factors could spur the development of gene-targeting applications in non-dividing cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • BRCA1 Protein / metabolism
  • BRCA2 Protein / metabolism
  • CRISPR-Cas Systems / genetics
  • Carrier Proteins / metabolism
  • Cell Line
  • Cullin Proteins / metabolism
  • DNA / metabolism
  • DNA Damage
  • DNA Repair
  • Fanconi Anemia Complementation Group N Protein
  • G1 Phase*
  • G2 Phase
  • Gene Targeting
  • Homologous Recombination*
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Kelch-Like ECH-Associated Protein 1
  • Molecular Sequence Data
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / metabolism
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism
  • Protein Binding
  • Rad51 Recombinase / metabolism
  • S Phase
  • Thiolester Hydrolases / metabolism
  • Tumor Suppressor Proteins / chemistry
  • Tumor Suppressor Proteins / metabolism
  • Ubiquitin-Protein Ligases / metabolism
  • Ubiquitination

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • BRCA2 Protein
  • BRCA2 protein, human
  • CUL3 protein, human
  • Carrier Proteins
  • Cullin Proteins
  • Fanconi Anemia Complementation Group N Protein
  • Intracellular Signaling Peptides and Proteins
  • KEAP1 protein, human
  • Kelch-Like ECH-Associated Protein 1
  • Multiprotein Complexes
  • Nuclear Proteins
  • PALB2 protein, human
  • RBX1 protein, human
  • Tumor Suppressor Proteins
  • USP11 protein, human
  • DNA
  • Ubiquitin-Protein Ligases
  • RAD51 protein, human
  • Rad51 Recombinase
  • Thiolester Hydrolases