期刊
出版社
ELSEVIER
DOI: 10.1016/j.mrfmmm.2020.111727
关键词
PNKP; The FHA domain; DNA repair; Genome stability
资金
- Uehara Memorial Foundation
- Takeda Science Foundation
- Kato Memorial Bioscience Foundation
- Japan Atomic Energy Agency
- Chubu Electric Power
- Tokyo Tech Academy for Leadership
- Tokyo Tech Academy for Cocreative Education of Environment and Energy Science
- Japan Society for the Promotion of Science [JP15H02817, JP17K20042, JP20H04334, JP18K11642]
- Radiation Effects Association
- [JP20J13601]
PNKP is recruited to DNA damage sites via interactions between its FHA domain and XRCC1 or XRCC4, playing a crucial role in DNA repair and maintenance of genome stability.
Polynucleotide kinase phosphatase (PNKP) has dual enzymatic activities as kinase and phosphatase for DNA ends, which are the prerequisite for the ligation, and thus is involved in base excision repair, single-strand break repair and non-homologous end joining for double-strand break (DSB) repair. In this study, we examined mechanisms for the recruitment of PNKP to DNA damage sites by laser micro-irradiation and live-cell imaging analysis using confocal microscope. We show that the forkhead-associated (FHA) domain of PNKP is essential for the recruitment of PNKP to DNA damage sites. Arg35 and Arg48 within the FHA domain are required for interactions with XRCC1 and XRCC4. PNKP R35A/R48A mutant failed to accumulate on the laser track and siRNAmediated depletion of XRCC1 and/or XRCC4 reduced PNKP accumulation on the laser track, indicating that PNKP is recruited to DNA damage sites via the interactions between its FHA domain and XRCC1 or XRCC4. Furthermore, cells expressing PNKP R35A/R48A mutant exhibited increased sensitivity toward ionizing radiation in association with delayed SSB and DSB repair and genome instability, represented by micronuclei and chromosome bridges. Taken together, these findings revealed the importance of PNKP recruitment to DNA damage sites via its FHA domain for DNA repair and maintenance of genome stability.
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