ALC1/CHD1L, a chromatin-remodeling enzyme, is required for efficient base excision repair

IR HANDLE Open Access
  • Tsuda, Masataka
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University
  • Cho, Kosai
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto Universit･Department of Primary Care and Emergency Medicine, Kyoto University Graduate School of Medicine
  • Ooka, Masato
    Department of Chemistry, Tokyo Metropolitan University
  • Shimizu, Naoto
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University
  • Watanabe, Reiko
    National Institutes of Biomedical Innovation, Health and Nutrition, Osaka
  • Yasui, Akira
    Division of Dynamic Proteome, Institute of Development, Aging and Cancer, Tohoku University
  • Nakazawa, Yuka
    Department of Genome Repair, Atomic Bomb Disease Institute, Nagasaki University Sakamoto･Department of Genetics, Research Institute of Environmental Medicine (RIeM), Nagoya University
  • Ogi, Tomoo
    Department of Genome Repair, Atomic Bomb Disease Institute, Nagasaki University Sakamoto･Department of Genetics, Research Institute of Environmental Medicine (RIeM), Nagoya University
  • Harada, Hiroshi
    Laboratory of Cancer Cell Biology, Radiation Biology Center, Kyoto University
  • Agama, Keli
    Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda
  • Nakamura, Jun
    Department of Environmental Sciences and Engineering, University of North Carolina Chapel Hill
  • Asada, Ryuta
    Department of Chemistry, Tokyo Metropolitan University
  • Fujiike, Haruna
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University
  • Sakuma, Tetsushi
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
  • Yamamoto, Takashi
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
  • Murai, Junko
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University･Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda
  • Hiraoka, Masahiro
    Department of Radiation Oncology, Japanese Red Cross Society Wakayama Medical Center
  • Koike, Kaoru
    Department of Primary Care and Emergency Medicine, Kyoto University Graduate School of Medicine
  • Pommier, Yves
    Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda
  • Takeda, Shunichi
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University
  • Hirota, Kouji
    Department of Radiation Genetics, Graduate School of Medicine, Kyoto University･Department of Chemistry, Tokyo Metropolitan University

Abstract

ALC1/CHD1L is a member of the SNF2 superfamily of ATPases carrying a macrodomain that binds poly(ADP-ribose). Poly(ADP-ribose) polymerase (PARP) 1 and 2 synthesize poly(ADP-ribose) at DNA-strand cleavage sites, promoting base excision repair (BER). Although depletion of ALC1 causes increased sensitivity to various DNA-damaging agents H₂O₂, UV, and phleomycin), the role played by ALC1 in BER has not yet been established. To explore this role, as well as the role of ALC1’s ATPase activity in BER, we disrupted the ALC1 gene and inserted the ATPase-dead (E165Q) mutation into the ALC1 gene in chicken DT40 cells, which do not express PARP2. The resulting ALC1⁻/⁻ and ALC1⁻/E165Q cells displayed an indistinguishable hypersensitivity to methylmethane sulfonate (MMS), an alkylating agent, and to H₂O₂, indicating that ATPase plays an essential role in the DNA-damage response. PARP1⁻/⁻ and ALC⁻/⁻/PARP1⁻/⁻ cells exhibited a very similar sensitivity to MMS, suggesting that ALC1 and PARP1 collaborate in BER. Following pulse-exposure to H₂O₂, PARP1⁻/⁻ and ALC1⁻/⁻/PARP1⁻/⁻ cells showed similarly delayed kinetics in the repair of single-strand breaks, which arise as BER intermediates. To ascertain ALC1’s role in BER in mammalian cells, we disrupted the ALC1 gene in human TK6 cells. Following exposure to MMS and to H₂O₂, the ALC⁻/⁻ TK6 cell line showed a delay in single-strand-break repair. We therefore conclude that ALC1 plays a role in BER. Following exposure to H₂O₂, ALC⁻/⁻ cells showed compromised chromatin relaxation. We thus propose that ALC1 is a unique BER factor that functions in a chromatin context, most likely as a chromatin-remodeling enzyme.

Journal

  • PloS one

    PloS one 12 (11), e0188320-, 2017-11-17

    Public Library of Science (PLoS)

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