• Open Access

Artemis-dependent DNA double-strand break formation at stalled replication forks

Authors

  • Junya Unno,

    1. Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
    2. Laboratory of DNA damage signaling, Department of Late Effect Studies, Radiation Biology Center, Kyoto University, Kyoto, Japan
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  • Masatoshi Takagi,

    Corresponding author
    • Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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  • Jinhua Piao,

    1. Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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  • Masataka Sugimoto,

    1. Section of Biochemistry, Department of Mechanism of Aging, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, Aichi, Japan
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  • Fumiko Honda,

    1. Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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  • Daisuke Maeda,

    1. Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan
    2. ADP-ribosylation in Oncology Project, National Cancer Center Research Institute, Tokyo, Japan
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  • Mitsuko Masutani,

    1. Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan
    2. ADP-ribosylation in Oncology Project, National Cancer Center Research Institute, Tokyo, Japan
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  • Tohru Kiyono,

    1. Virology Division, National Cancer Center Research Institute, Tokyo, Japan
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  • Fumiaki Watanabe,

    1. Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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  • Tomohiro Morio,

    1. Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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  • Hirobumi Teraoka,

    1. Department of Pathological Biochemistry, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan
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  • Shuki Mizutani

    Corresponding author
    • Department of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan
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To whom correspondence should be addressed.

E-mails: m.takagi.ped@tmd.ac.jp or smizutani.ped@tmd.ac.jp

Abstract

Stalled replication forks undergo DNA double-strand breaks (DSBs) under certain conditions. However, the precise mechanism underlying DSB induction and the cellular response to persistent replication fork stalling are not fully understood. Here we show that, in response to hydroxyurea exposure, DSBs are generated in an Artemis nuclease-dependent manner following prolonged stalling with subsequent activation of the ataxia–telangiectasia mutated (ATM) signaling pathway. The kinase activity of the catalytic subunit of the DNA-dependent protein kinase, a prerequisite for stimulation of the endonuclease activity of Artemis, is also required for DSB generation and subsequent ATM activation. Our findings indicate a novel function of Artemis as a molecular switch that converts stalled replication forks harboring single-stranded gap DNA lesions into DSBs, thereby activating the ATM signaling pathway following prolonged replication fork stalling.

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