ATR-ATRIP kinase complex triggers activation of the fanconi anemia DNA repair pathway

Tomoko Shigechi, Junya Tomida, Koichi Sato, Masahiko Kobayashi, John K. Eykelenboom, Fabio Pessina, Yanbin Zhang, Emi Uchida, Masamichi Ishiai, Noel F. Lowndes, Kenichi Yamamoto, Hitoshi Kurumizaka, Yoshihiko Maehara, Minoru Takata

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    Abstract

    ATR kinase activates the S-phase checkpoint when replication forks stall at sites of DNA damage. This event also causes phosphorylation of the Fanconi anemia (FA) protein FANCI, triggering its monoubiquitination of the key DNA repair factor FANCD2 by the FA core E3 ligase complex, thereby promoting this central pathway of DNA repair which permits replication to be restarted. However, the interplay between ATR and the FA pathway has been unclear. In this study, we present evidence that their action is directly linked, gaining insights into this relationship in a DT40 mutant cell line that is conditionally deficient in the critical ATR-binding partner protein ATRIP. Using this system, we showed that ATRIP was crucial for DNA damage-induced FANCD2 monoubiquitination and FANCI phosphorylation. ATR kinase phosphorylated recombinant FANCI protein in vitro, which was facilitated by the presence of FANCD2. Mechanistic investigations revealed that the RPA region but not the TopBP1 region of ATRIP was required for FANCD2 monoubiquitination, whereas Chk1 phosphorylation relied upon both domains. Together, our findings identify ATR as the kinase responsible for activating the FA pathway of DNA repair.

    Original languageEnglish
    Pages (from-to)1149-1156
    Number of pages8
    JournalCancer Research
    Volume72
    Issue number5
    DOIs
    Publication statusPublished - 2012 Mar 1

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    ASJC Scopus subject areas

    • Cancer Research
    • Oncology

    Cite this

    Shigechi, T., Tomida, J., Sato, K., Kobayashi, M., Eykelenboom, J. K., Pessina, F., Zhang, Y., Uchida, E., Ishiai, M., Lowndes, N. F., Yamamoto, K., Kurumizaka, H., Maehara, Y., & Takata, M. (2012). ATR-ATRIP kinase complex triggers activation of the fanconi anemia DNA repair pathway. Cancer Research, 72(5), 1149-1156. https://doi.org/10.1158/0008-5472.CAN-11-2904