Replication stress induces accumulation of FANCD2 at central region of large fragile genes

Yusuke Okamoto, Watal M. Iwasaki, Kazuto Kugou, Kazuki K. Takahashi, Arisa Oda, Koichi Sato, Wataru Kobayashi, Hidehiko Kawai, Ryo Sakasai, Akifumi Takaori-Kondo, Takashi Yamamoto, Masato T. Kanemaki, Masato Taoka, Toshiaki Isobe, Hitoshi Kurumizaka, Hideki Innan, Kunihiro Ohta, Masamichi Ishiai, Minoru Takata

    Research output: Contribution to journalArticle

    17 Citations (Scopus)

    Abstract

    During mild replication stress provoked by low dose aphidicolin (APH) treatment, the key Fanconi anemia protein FANCD2 accumulates on common fragile sites, observed as sister foci, and protects genome stability. To gain further insights into FANCD2 function and its regulatory mechanisms, we examined the genome-wide chromatin localization of FANCD2 in this setting by ChIP-seq analysis. We found that FANCD2 mostly accumulates in the central regions of a set of large transcribed genes that were extensively overlapped with known CFS. Consistent with previous studies, we found that this FANCD2 retention is R-loop-dependent. However, FANCD2 monoubiquitination and RPA foci formation were still induced in cells depleted of R-loops. Interestingly, we detected increased Proximal Ligation Assay dots between FANCD2 and R-loops following APH treatment, which was suppressed by transcriptional inhibition. Collectively, our data suggested that R-loops are required to retain FANCD2 in chromatin at the middle intronic region of large genes, while the replication stress-induced upstream events leading to the FA pathway activation are not triggered by R-loops.

    Original languageEnglish
    Pages (from-to)2932-2944
    Number of pages13
    JournalNucleic Acids Research
    Volume46
    Issue number6
    DOIs
    Publication statusPublished - 2018 Jan 1

    ASJC Scopus subject areas

    • Genetics

    Fingerprint Dive into the research topics of 'Replication stress induces accumulation of FANCD2 at central region of large fragile genes'. Together they form a unique fingerprint.

  • Cite this

    Okamoto, Y., Iwasaki, W. M., Kugou, K., Takahashi, K. K., Oda, A., Sato, K., Kobayashi, W., Kawai, H., Sakasai, R., Takaori-Kondo, A., Yamamoto, T., Kanemaki, M. T., Taoka, M., Isobe, T., Kurumizaka, H., Innan, H., Ohta, K., Ishiai, M., & Takata, M. (2018). Replication stress induces accumulation of FANCD2 at central region of large fragile genes. Nucleic Acids Research, 46(6), 2932-2944. https://doi.org/10.1093/nar/gky058