ALC1/CHD1L, chromatin-remodeling enzyme, is required for efficient base excision repair
Autor: | Tetsushi Sakuma, Kouji Hirota, Yuka Nakazawa, Haruna Fujiike, Naoto Shimizu, Keli Agama, Shunichi Takeda, Jun Nakamura, Junko Murai, Masataka Tsuda, Tomoo Ogi, Akira Yasui, Yves Pommier, Takashi Yamamoto, Ryuta Asada, Masato Ooka, Reiko Watanabe, Hiroshi Harada, Kaoru Koike, Kosai Cho, Masahiro Hiraoka |
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Jazyk: | angličtina |
Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Adenosine Triphosphatase DNA Repair Poly (ADP-Ribose) Polymerase-1 lcsh:Medicine Gene Expression Biochemistry Mechanical Treatment of Specimens CHD1L Poultry Histones 0302 clinical medicine PARP1 Gamefowl lcsh:Science Cell Disruption Cell Line Transformed B-Lymphocytes Multidisciplinary biology Chemistry Chromosome Biology Eukaryota Base excision repair Chromatin Cell biology Enzymes Nucleic acids DNA-Binding Proteins Histone Specimen Disruption 030220 oncology & carcinogenesis Vertebrates Epigenetics Poly(ADP-ribose) Polymerases Research Article Signal Transduction DNA repair Poly ADP ribose polymerase Research and Analysis Methods Chromatin remodeling Birds 03 medical and health sciences Cell Line Tumor Genetics Animals Gene Disruption Humans Molecular Biology Techniques Molecular Biology Osteoblasts lcsh:R Organisms Phosphatases DNA Helicases Biology and Life Sciences Proteins Cell Biology DNA Hydrogen Peroxide Chromatin Assembly and Disassembly Methyl Methanesulfonate 030104 developmental biology Gene Expression Regulation Fowl Specimen Preparation and Treatment Amniotes biology.protein Enzymology DNA damage lcsh:Q Chickens Cloning HeLa Cells |
Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 11, p e0188320 (2017) |
Popis: | 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 ALC1-/-/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 ALC1-/- 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, ALC1-/- 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. This work was supported by the JSPS KAKENHI Grant Number (JP16K12598, JP16H02957 and JP16H01314 to KH, JP16H06306 to ST and JP16J02252 to RA), the JSPS Core-to-Core Program (A) Advanced Research Networks (to ST), the Takeda Science Foundation and Yamada Science Foundation (to KH), and the Center for Cancer Research, Intramural Program of the US National Cancer Institute (Z01 BC 006150 to KA and YP). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. |
Databáze: | OpenAIRE |
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