RFWD3-Mediated Ubiquitination Promotes Timely Removal of Both RPA and RAD51 from DNA Damage Sites to Facilitate Homologous Recombination.

RFWD3 is a recently identified Fanconi anemia protein FANCW whose E3 ligase activity toward RPA is essential in homologous recombination (HR) repair. However, how RPA ubiquitination promotes HR remained unknown. Here, we identified RAD51, the central HR protein, as another target of RFWD3. We show that RFWD3 polyubiquitinates both RPA ...
and RAD51 in vitro and in vivo. Phosphorylation by ATR and ATM kinases is required for this activity in vivo. RFWD3 inhibits persistent mitomycin C (MMC)-induced RAD51 and RPA foci by promoting VCP/p97-mediated protein dynamics and subsequent degradation. Furthermore, MMC-induced chromatin loading of MCM8 and RAD54 is defective in cells with inactivated RFWD3 or expressing a ubiquitination-deficient mutant RAD51. Collectively, our data reveal a mechanism that facilitates timely removal of RPA and RAD51 from DNA damage sites, which is crucial for progression to the late-phase HR and suppression of the FA phenotype.
Mesh Terms:
Adenosine Triphosphatases, Ataxia Telangiectasia Mutated Proteins, Binding Sites, Cell Cycle Proteins, Cell Line, Tumor, Chromatin, DNA, DNA Damage, Fanconi Anemia, Humans, Minichromosome Maintenance Proteins, Mitomycin, Mutation, Phosphorylation, Proteasome Endopeptidase Complex, Protein Binding, Proteolysis, RNA Interference, Rad51 Recombinase, Recombinational DNA Repair, Replication Protein A, Transfection, Ubiquitin-Protein Ligases, Ubiquitination, Valosin Containing Protein
Mol. Cell
Date: Jun. 01, 2017
Download Curated Data For This Publication
207192
Switch View:
  • Interactions 10