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.
Pubmed ID: 28575658 RIS Download
Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.
A commercial graphing software company that offers scientific software for statistical analyses, curve fitting and data analysis. It offers four programs: Prism, InStat, StatMate and QuickCalcs.
View all literature mentionsThis monoclonal targets Histone H3
View all literature mentionsThis monoclonal targets Chk1 (G-4)
View all literature mentionsThis monoclonal targets RPA70
View all literature mentionsThis monoclonal targets FLAG??
View all literature mentionsThis polyclonal targets FANCD2
View all literature mentionsThis monoclonal targets DMC1
View all literature mentionsThis monoclonal targets c-Myc
View all literature mentionsThis monoclonal targets 6X His tag??
View all literature mentionsThis monoclonal targets α-tubulin
View all literature mentionsThis monoclonal targets Green Fluorescent Protein
View all literature mentionsThis monoclonal targets anti-DDDDK-tag
View all literature mentionsThis monoclonal targets Chk1, phospho (Ser345)
View all literature mentionsThis polyclonal targets RPA3 antibody produced in rabbit
View all literature mentionsThis unknown targets Phospho-RPA32 (Ser33)
View all literature mentionsThis unknown targets Phospho-RPA32 (Ser4, Ser8)
View all literature mentionsThis monoclonal targets RPA32/RPA2
View all literature mentionsThis polyclonal targets RFWD3 antibody
View all literature mentionsThis monoclonal targets Chk1 (G-4)
View all literature mentions