Genomic mapping of radiation damage using a next generation sequencer

• Project/Area Number: 26550030
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Hiroshima University
• Principal Investigator: Hiroshi IDE 広島大学, 理学(系)研究科(研究院), 教授 (30223126)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 放射線 / DNA損傷 / DNA修復

Outline of Final Research Achievements

DNA-protein cross-links (DPCs) are major DNA lesions induced by ionizing radiation and anticancer drugs. However, it remains elusive whether some regions of the genome are more vulnerable with respect to DPC formation than others when cells are exposed to DNA damaging agents. In the present study, we developed a method for genomic mapping of radiation-induced DPCs. Mouse SCCVII tumors were irradiated with X-rays and DNA was purified. Proteins present in DPCs were labeled with biotin and DNA fragments containing DPCs (DPC-DNA) were specifically pulled down using avidin-beads. A synthetic adaptor was ligated at both ends of DPC-DNA. The resulting DNA was subjected to PCR. Analysis of reaction products with agarose gel electrophoresis showed that DPC-DNA was successfully amplified by PCR.

Research Products

• [Journal Article] Role of isolated and clustered DNA damage and the post-irradiating repair process in the effects of heavy ion beam irradiation (2015)
  • Author(s): Yuka Tokuyama, Yoshiya Furusawa, Hiroshi Ide, Akira Yasui, Hiroaki Terato
  • Journal Title: Journal of Radiation Research Volume: 56 Issue: 3 Pages: 000-000
  • DOI: 10.1093/jrr/rru122
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Induction of DNA-protein cross-links by ionizing radiation and their repair in cells (2016)
  • Author(s): Ide H, Sebata T, Kuboyama M, Xu X, Nakano T, Salem A, Shoulkamy M, Hirayama R, Uzawa A.
  • Organizer: 14th International Workshop on Radiation Damage to DNA
  • Place of Presentation: Victorian Treasury Theatre, Melbourne, Australia
  • Year and Date: 2016-03-20
  • Int'l Joint Research / Invited
• [Presentation] DNA-タンパク質クロスリンク損傷の新規な定量法 (2015)
  • Author(s): 瀬畑敬文，久保山政弥，中野敏彰，徐徐，Salem Amir，Shoulkamy Mahmoud，平山亮一，鵜澤玲子，井出博
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸国際展示場，神戸
  • Year and Date: 2015-12-01
• [Presentation] 種々のDNA傷害因子が誘発するDNA-タンパク質クロスリンク損傷の解析 (2015)
  • Author(s): 久保山政弥，瀬畑敬文，謝明章，合田美月，Shoulkamy Mahmoud，Salem Amir，井出博，中野敏彰
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸国際展示場，神戸
  • Year and Date: 2015-12-01
• [Presentation] X線照射したマウス腫瘍におけるDNA-タンパク質クロスリンクの生成・除去動態 (2015)
  • Author(s): 中野敏彰，瀬畑敬文，久保山政弥，徐徐，Salem Amir，Shoulkamy Mahmoud，平山亮一, 鵜澤玲子，井出博
  • Organizer: 日本放射線影響学会 第1回放射線ワークショップ
  • Place of Presentation: 富山大学黒田講堂，富山
  • Year and Date: 2015-10-16
• [Presentation] Induction of DNA-protein cross-link damage by ionizing radiation and its elimination from the genome (2015)
  • Author(s): Ide H, Nakano T, Sebata T, Kuboyama M, Hirayama R, Uzawa A, Furusawa Y.
  • Organizer: 15th International Congress of Radiation Research
  • Place of Presentation: Kyoto International Conference Center, Kyoto, Japan
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] アルデヒドが誘発するDNA損傷の解析 (2014)
  • Author(s): 中野敏彰，謝明章，合田美月，Mahmoud Shoulkamy，井出博
  • Organizer: 日本環境変異原学会第43回大会
  • Place of Presentation: 一橋大学一橋講堂，東京
  • Year and Date: 2014-12-04 – 2014-12-05
• [Presentation] Analysis of DNA-protein cross-link damage induced by ionizing radiation (2014)
  • Author(s): Amir Salem，服部峻，瀬畑敬文，久保山政弥，中野敏彰，井出博
  • Organizer: 第39回中国地区放射線影響研究会
  • Place of Presentation: 広島大学医学部，広島市
  • Year and Date: 2014-07-18