Regulation of 53BP1 repositioning and foci formation during homologous recombination

• Project/Area Number: 17K12821
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Nagoya University (2019); Nagasaki University (2018); Sasaki Foundation (2017)
• Principal Investigator: ISONO Mayu 名古屋大学, 環境医学研究所, 特任助教 (90713511)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: DNA二本鎖切断 / DSB修復 / 53BP1 / ATR活性 / DSB修復経路 / 53BP1 repositioning / DSB修復経路選択 / ヒストン修飾

Outline of Final Research Achievements

DNA double-strand break (DSB) caused by ionizing radiation or anti-cancer drugs is a deleterious damage, which can lead to chromosomal instability. Cells have the two main repair pathways for DSB, non-homologous end-joining (NHEJ) and homologous recombination (HR). While NHEJ functions through cell cycle, HR ensues if NHEJ fails to repair DSB in S/G2 phase. 53BP1 is known as a factor promoting NHEJ via inhibiting HR. We show the activation of ATM and ATR is dispensable for the maintenance of 53BP1 foci at sites of DSBs generated by ionizing radiation at early time points (0.5 h). Interestingly, inhibition of ATR (but not ATM) disturbs the maintenance of 53BP1 foci at later time points (4 h), which is fully recovered when the inhibitor is omitted. Collectively, the results in this study suggest that localization of 53BP1 foci at DSB sites during HR is dependent on ATR activation.

Academic Significance and Societal Importance of the Research Achievements

今回の研究結果から、HR進行中のDSB部位における53BP1の集積の維持には、DSB誘発早期に活性化するATMではなくresectionの開始に伴い活性化するATRに依存することが示唆された。HR修復における53BP1の役割に関して、今後より詳細な解析を進めることができれば、発がん等の疾病発症のメカニズム解明の一助になると考えている。

Research Products

• [Journal Article] Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair (2020)
  • Author(s): Nakazawa Y, Hara Y, Oka Y, Komine O, van den Heuvel D, Guo C, Daigaku Y, Isono M, He Y, Shimada M, Kato K, Jia N, Hashimoto S, Kotani Y, Miyoshi Y, Tanaka M, Sobue A, Mitsutake N, Suganami T, Masuda A, Ohno K, Nakada S, Mashimo T, Yamanaka K, Luijsterburg MS, Ogi T
  • Journal Title: Cell Volume: 19 Issue: 6 Pages: 1228-1244
  • DOI: 10.1016/j.cell.2020.02.010
  • NAID: 120006871293
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] DNA Fragment Agarose Gel Electrophoresis for Chromatin Immunoprecipitation (ChIP) (2020)
  • Author(s): Isono M, Hashimoto S.
  • Journal Title: Methods in Molecular Biology Volume: 2119 Pages: 201-211
  • DOI: 10.1007/978-1-0716-0323-9_17
  • ISBN: 9781071603222, 9781071603239
  • Peer Reviewed
• [Journal Article] DNA二本鎖切断修復経路を最適化する時空間的制御機構 (2018)
  • Author(s): 磯野真由, 柴田淳史
  • Journal Title: 放射線生物研究 Volume: 53 Pages: 116-131
  • NAID: 40021642264
  • Peer Reviewed / Open Access
• [Presentation] 転写共役ヌクレオチド除去修復機構に重要なRNAポリメラーゼユビキチン化部位の同定. (2019)
  • Author(s): 中沢由華, 原雄一郎, 岡泰由, 小峯起, Diana van den Heuvel, 郭朝万, 大学保一, 磯野真由, 何予希, 嶋田繭子, 加藤香奈, 賈楠, 橋下悟, 小谷祐子, 三好由夏, 田中都, 祖父江顕, 光武範吏, 菅波孝祥, 増田章男, 大野欽司, 中田慎一郎, 真下知士, 山中宏二, Martijn S. Luijsterburg, 荻朋男.
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] ChIP-seqを利用したDNA損傷およびヌクレオチド除去修復のモニタリング. (2019)
  • Author(s): 原雄一郎, 中沢由華, 岡泰由, 小峯起, Diana van den Heuvel, 郭朝万, 大学保一, 磯野真由, 何予希, 嶋田繭子, 加藤香奈, 賈楠, 橋下悟, 小谷祐子, 三好由夏, 田中都, 祖父江顕, 光武範吏, 菅波孝祥, 増田章男, 大野欽司, 中田慎一郎, 真下知士, 山中宏二, Martijn S. Luijsterburg, 荻朋男.
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] 相同組換え修復への経路切り替えを促進する時空間的53BP1動態制御機構 (2018)
  • Author(s): 磯野真由, 萩原慶彦, 佐藤浩央, 尾池貴洋, 西良太郎, 中田慎一郎, 中野隆史, 鈴木啓司, 柴田淳史
  • Organizer: 日本放射線影響学会第61回大会