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

Shojiro Inano, Koichi Sato, Yoko Katsuki, Wataru Kobayashi, Hiroki Tanaka, Kazuhiro Nakajima, Shinichiro Nakada, Hiroyuki Miyoshi, Kerstin Knies, Akifumi Takaori-Kondo, Detlev Schindler, Masamichi Ishiai, Hitoshi Kurumizaka, Minoru Takata

    Research output: Contribution to journalArticle

    42 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)622-634.e8
    JournalMolecular Cell
    Volume66
    Issue number5
    DOIs
    Publication statusPublished - 2017 Jun 1

    Keywords

    • BRCA2
    • Fanconi anemia
    • homologous recombination
    • ICL repair
    • RAD51
    • RFWD3
    • RPA

    ASJC Scopus subject areas

    • Molecular Biology
    • Cell Biology

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  • Cite this

    Inano, S., Sato, K., Katsuki, Y., Kobayashi, W., Tanaka, H., Nakajima, K., Nakada, S., Miyoshi, H., Knies, K., Takaori-Kondo, A., Schindler, D., Ishiai, M., Kurumizaka, H., & Takata, M. (2017). RFWD3-Mediated Ubiquitination Promotes Timely Removal of Both RPA and RAD51 from DNA Damage Sites to Facilitate Homologous Recombination. Molecular Cell, 66(5), 622-634.e8. https://doi.org/10.1016/j.molcel.2017.04.022