Visualization of heavy-ion induced clustered DNA double strand break formation using super-resolution microscopy

• Project/Area Number: 15K15449
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Radiation science
• Research Institution: Sasaki Foundation (2016); Gunma University (2015)
• Principal Investigator: Isono Mayu 公益財団法人佐々木研究所, 附属研究所, 研究員(移行) (90713511)
• Research Collaborator: SHIBATA Atsushi 国立大学法人群馬大学, 医学系研究科, 助教 (30707633)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: DNAクラスター損傷 / 重粒子線 / 染色体異常 / DNA修復 / ゲノム不安定性 / 超高解像度顕微鏡

Outline of Final Research Achievements

Heavy-ion radiation induces clustered DNA damage, which contains multiple type of DNA damage, e.g. DNA double strand break (DSB), single strand break and base damage. However so far, such clustered DNA damage has not been visualized in the cellular level. Here we aimed to visualize the clustered DNA damage, particularly about the formation of DSB, after heavy-ion radiation by using 3D super-resolution microscopy. To address the question whether heavy-ion irradiation generates clustered DSB formation or not, we monitored RPA foci as a DSB marker. The super-resolution analysis revealed that heavy-ion radiation induced multiple RPA foci in close proximity. Furthermore, our analysis demonstrated that the distance between two individual RPA foci within γH2AX foci was approximately 700 nm. In summary, our data suggests that these closely localized DSBs are considered to be a risk for chromosomal translocations, deletions and cell death after heavy-ion irradiation.

Research Products

• [Journal Article] BRCA1 Directs the Repair Pathway to Homologous Recombination by Promoting 53BP1 Dephosphorylation (2017)
  • Author(s): Mayu Isono, Atsuko Niimi, Takahiro Oike, Yoshihiko Hagiwara, Hiro Sato, Ryota Sekine, Yukari Yoshida, Shin-Ya Isobe, Chikashi Obuse, Ryotaro Nishi, Elena Petricci, Shinichiro Nakada, Takashi Nakano, Atsushi Shibata
  • Journal Title: Cell Rep. Volume: 18 Pages: 520-532
  • DOI: 10.1016/j.celrep.2016.12.042
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Carbon-ion beams effectively induce growth inhibition and apoptosis in human neural stem cells compared with glioblastoma A172 cells (2015)
  • Author(s): Isono M, Yoshida Y, Takahashi A, Oike T, Shibata A, Kubota Y, Kanai T, Ohno T, Nakano T.
  • Journal Title: J Radiat Res Volume: 56 Pages: 856-861
  • DOI: 10.1093/jrr/rrv033
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The EGFR mutation status affects the relative biological effectiveness of carbon-ion beams in non-small cell lung carcinoma cells (2015)
  • Author(s): Amornwichet N, Oike T, Shibata A, Nirodi CS, Ogiwara H, Makino H, Kimura Y, Hirota Y, Isono M, Yoshida Y, Ohno T, Kohno T, Nakano T.
  • Journal Title: Scientific Reports Volume: 11 Pages: 11305-11305
  • DOI: 10.1038/srep11305
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] 炭素イオン線誘発DNA二本鎖切断の修復経路選択における線エネルギー付与の効果 (2016)
  • Author(s): 萩原慶彦，磯野真由，新美敦子，尾池貴洋，佐藤浩央，中野隆史，柴田淳史
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: 横浜市, 神奈川県
  • Year and Date: 2016-11-30
• [Presentation] BRCA1 directs the repair pathway to homologous recombination by promoting 53BP1 dephosphorylation (2016)
  • Author(s): Mayu Isono, Atsuko Niimi, Yoshihiko Hagiwara, Ryotaro Nishi, Shinya Isobe, Chikashi Obuse, Elena Petricci, Shinichiro Nakada, Takashi Nakano and Atsushi Shibata
  • Organizer: The 10th 3R Symposium
  • Place of Presentation: Matsue, Shimane, Japan
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] The effect of Linear Energy Transfer in DNA double strand break repair pathway choice in G2 cells (2016)
  • Author(s): Yoshihiko Hagiwara, Atsuko Niimi, Mayu Isono, Takahiro Oike, Hiro Sato, Takashi Nakano and Atsushi Shibata
  • Organizer: The 10th 3R Symposium
  • Place of Presentation: Matsue, Shimane, Japan
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] BRCA1によるDSB end resectionの時空間的制御 (2015)
  • Author(s): 磯野真由
  • Organizer: 第38回 日本分子生物学会・第88回日本生化学会 合同大会
  • Place of Presentation: 神戸ポートピアホテル 本館 B1F 偕楽1(神戸市)
  • Year and Date: 2015-12-03
• [Presentation] The complexity of damage at the DNA break ends induces rapid DNA double strand break resection (2015)
  • Author(s): 磯野真由
  • Organizer: ICRR2015
  • Place of Presentation: 国立京都国際会議場(京都市)
  • Year and Date: 2015-05-25
  • Int'l Joint Research