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
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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