Role of protein phosphorylating function of DNA-PK in DNA double-strand break repair

• Project/Area Number: 15H02817
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Matsumoto Yoshihisa 東京工業大学, 科学技術創成研究院, 准教授 (20302672)
• Research Collaborator: SHIMADA MIKIO ; SHARMA MUKESH KUMAR ; AMIRI MOGHAHANI ALI REZA ; ASA ANIE DAY DE CASTRO
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000); Fiscal Year 2017: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2016: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2015: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000)
• Keywords: 放射線 / がん / DNA / DNA修復 / タンパク質リン酸化 / シグナル伝達 / ＤＮＡ / 癌治療 / 発がん / DNA損傷応答 / 抗体 / 放射線高感受性遺伝病

Outline of Final Research Achievements

This study aimed to clarify the substrate and role of protein phosphorylation by DNA-PK, which is considered the molecular sensor for DNA double-strand break (DSB). Regarding XRCC4, which is involved in the joining reaction of two DNA ends at the final step of DSB repair, we generated the phosphorylation-specific antibodies, corresponding to six phosphorylation sites identified by us and other groups. By using these antibodies, we clarified the response of phosphorylation to radiation. Through the generation and analyses of phosphorylation-disabled mutants, we also clarified the phosphorylation sites, which are essential for DSB repair. Additionally, through the generation and analyses of XRCC4 mutants, mimicking the mutations found in patients of microcephaly and growth defects, we clarified the impact of disease-associated mutations on the function of XRCC4 in DSB repair.

Academic Significance and Societal Importance of the Research Achievements

DNA二重鎖切断(DSB)は、放射線によって生じる種々のDNA損傷の中で最も重篤であり、がん放射線治療の成否の鍵を握ると考えられている。DNA-PKはDSBの「センサー」として注目されてきたが、DSB修復過程において、どのタンパク質を、何のためにリン酸化するかは20年以上未解明のままであった。本研究で、DSBの修復過程の最終段階でのDNA末端の結合に関わるXRCC4のリン酸化の放射線に対する応答、DSB修復における重要性が明らかになった。本研究の成果は、DSB修復過程の理解の深化とともに、がん放射線治療における放射線感受性の予測や増感剤開発などに示唆を与えるものである。

Research Products

• [Int'l Joint Research] SPC Government College(インド)
• [Int'l Joint Research] Mahidol University(タイ)
• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Int'l Joint Research] サセックス大学(英国)
• [Int'l Joint Research] SPC Government College/ボパール記念病院・医学研究所(インド)
• [Int'l Joint Research] マヒドル大学(タイ)
• [Int'l Joint Research] CEA(フランス)
• [Int'l Joint Research]
• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Int'l Joint Research] サセックス大学(英国)
• [Int'l Joint Research] R.R. Government College/ボパール記念病院・医学研究所(インド)
• [Journal Article] Influence of XRCC4 expression by breast cancer cells on ipsilateral recurrence after breast-conserving therapy (2019)
  • Author(s): Kitagawa M, Someya M, Hasegawa T, Mikami T, Asaishi K, Hasegawa T, Matsumoto Y, Kutomi G, Takemasa I, Sakata K
  • Journal Title: Strahlentherapie und Oncologie Volume: - Issue: 7 Pages: 1-11
  • DOI: 10.1007/s00066-019-01468-z
  • Peer Reviewed
• [Journal Article] DNA damage response after ionizing radiation exposure in skin keratinocytes derived from human induced pluripotent stem cells (2019)
  • Author(s): Tomoko Miyake, Mikio Shimada, Yoshihisa Matsumoto, Akitoshi Okino
  • Journal Title: International Journal of Radiation Oncology, Biology, Physics Volume: -
  • Peer Reviewed
• [Journal Article] Structural analysis of DNA repair protein XRCC4 applying circular dichroism in an aqueous solution. (2018)
  • Author(s): Nishikubo K, Izumi Y, Matsumoto Y, Fujii K, Matsuo K, Yokoya A.
  • Journal Title: Radiat. Protect. Dosim. Volume: - Issue: 1-2 Pages: 1-4
  • DOI: 10.1093/rpd/ncy275
  • Peer Reviewed
• [Journal Article] In cellulo phosphorylation of DNA double-strand break repair protein XRCC4 on Ser260 by DNA-PK (2018)
  • Author(s): Amiri Moghani AR, Sharma MK, Matsumoto Y
  • Journal Title: Journal of Radiation Research Volume: 59 Pages: 700-708
  • DOI: 10.1093/jrr/rry072
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Caspase-mediated cleavage of X-ray repair cross-complementing group 4 promotes apoptosis by enhancing nuclear translocation of caspase- activated DNase (2018)
  • Author(s): 1.Sunatani Y, Kamdar RP, Sharma MK, Matsui T, Sakasai R, Hashimoto M, Ishigaki Y, Matsumoto Y, Iwabuchi K
  • Journal Title: Exp Cell Res Volume: 362 Issue: 2 Pages: 450-460
  • DOI: 10.1016/j.yexcr.2017.12.009
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Influence of XRCC4 expression in esophageal cancer cells on the response to radiotherapy. (2017)
  • Author(s): Hori M, Someya M, Matsumoto Y, Nakata K, Kitagawa M, Hasegawa T, Tsuchiya T, Fukushima Y, Gocho T, Sato Y, Ohnuma H, Kato J, Sugita S, Hasegawa T, Sakata K.
  • Journal Title: Medical Molecular Morphology Volume: 50 Issue: 1 Pages: 25-33
  • DOI: 10.1007/s00795-016-0144-5
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Local tumor control and DNA-PK activity of peripheral blood lymphocytes in prostate cancer patients receiving radiotherapy. (2016)
  • Author(s): Someya M, Hasegawa T, Hori M, Matsumoto Y, Nakata K, Masumori N, Sakata K.
  • Journal Title: Journal of Radiation Research Volume: 58 Issue: 2 Pages: 225-231
  • DOI: 10.1093/jrr/rrw099
  • Peer Reviewed / Open Access
• [Journal Article] Expression of Ku70 predicts results of radiotherapy in prostate cancer. (2016)
  • Author(s): Hasegawa T, Someya M, Hori M, Matsumoto Y, Nakata K, Nojima M, Kitagawa M, Tsuchiya T, Masumori N, Hasegawa T, Sakata K.
  • Journal Title: Strahlentherapie und Onkologie Volume: 193 Issue: 1 Pages: 29-37
  • DOI: 10.1007/s00066-016-1023-7
  • Peer Reviewed
• [Journal Article] LIG4 mediates Wnt signaling-induced radioresistance (2016)
  • Author(s): Jun S, Jung Y-S, Suh HN, Wang W, Kim MJ, Oh YS, Lien E, Shen X, Matsumoto Y, McCrea P, Li L, Chen J, *Park J-I.
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 10994-10994
  • DOI: 10.1038/ncomms10994
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage (2016)
  • Author(s): Sharma MK, Imamichi S, Fukuchi M, Samartha RM, Tomita M, Matsumoto Y.
  • Journal Title: Journal of Radiation Research Volume: 57 Issue: 2 Pages: 115-120
  • DOI: 10.1093/jrr/rrv086
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Influence of Ku86 and XRCC4 expression in uterine cervical cancer on the response to preoperative radiotherapy (2016)
  • Author(s): Takada Y, Someya M, Matsumoto Y, Satoh M, Nakata K, Hori M, Saito M, Horikawa N, Tateoka K, Teramoto M, Saito T, Hasegawa T, Sakata K.
  • Journal Title: Medical Molecular Morphology Volume: 印刷中
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] DNA-PKcs is involved in immunoglobulin class switch recombination in human B-cells (2015)
  • Author(s): Bjorkman A, Du L, Felgentreff K, Rosner C, Kamdar RP, Kokaraki G, Matsumoto Y, Davies EG, van der Burg M, Notarangelo LD, Hammarstrom L, Pan-Hammarstrom Q.
  • Journal Title: Journal of Immunology Volume: 195 Issue: 12 Pages: 5608-5615
  • DOI: 10.4049/jimmunol.1501633
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Regulation of DNA double-strand break repair via non-homologous end joining pathway (2018)
  • Author(s): Yoshihisa Matsumoto
  • Organizer: International Workshop on Influence of electromagnetic radiation on human beings, and problems of understanding
  • Int'l Joint Research / Invited
• [Presentation] Functional analysis of XRCC4 mutations associated with microcephaly and growth defects (2018)
  • Author(s): Anie Day Asa DC, Rujira Wanotayan, Mikio Shimada, Mukesh Kumar Sharma, Yoshihisa Matsumoto
  • Organizer: The 2nd International Symposium on Radiation Therapeutics and Biology & The 34th Radiation Biology Center International Symposium
  • Int'l Joint Research
• [Presentation] DNA damage-induced phosphorylation of XRCC4 on Ser260 and Ser320 by DNA-PK (2018)
  • Author(s): Ali Reza Amiri Moghani, Mukesh Kumar Sharma, Yoshihisa Matsumoto
  • Organizer: 3R&3C Symposium
  • Int'l Joint Research
• [Presentation] Functional Analysis of Disease-associated XRCC4 Mutations and Its Implication in the Development of Immune and Neural Systems (2018)
  • Author(s): Anie Day Asa De Castro, Rujira Wanotayan, Mukesh Kumar Sharma, Mikio Shimada, Yoshihisa Matsumoto
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] iPS 細胞におけるゲノム安定性維持に関わるリン酸化・脱リン酸化機構解析 (2018)
  • Author(s): 香川 望, 塚田 海馬, 島田 幹男, 松本 義久
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] In cellulo phosphorylation of DNA double-strand break repair protein XRCC4 on Ser260 by DNA-PK (2018)
  • Author(s): Ali Reza Amiri Moghani, Mukesh Kumar Sharma, Yoshihisa Matsumoto
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] VUV-CDを用いたXRCC4タンパク質の活性化二次構造解析 (2018)
  • Author(s): 西久保 開, 泉 雄大, 藤井 健太郎, 松尾 光一, 松本 義久, 横谷 明徳
  • Organizer: 量子生命科学研究会第2回学術集会
• [Presentation] DNA損傷応答タンパク質の活性化スイッチングとエピジェネティック構造変化の研究 (2018)
  • Author(s): 長谷川 真保, 西久保 開, 松本 義久, 藤原 悟, 横谷 明徳
  • Organizer: 平成30年度若手放射線生物学研究会専門研究会
• [Presentation] XRCC4 タンパク質の擬似リン酸化による構造変化の解析 (2018)
  • Author(s): 西久保 開, 長谷川 真保, 泉 雄大, 藤井 健太郎, 松尾 光一, 松本 義久, 横谷 明徳
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] DNA double-strand break repair and V(D)J recombination function of XRCC4 mutants associated with microcephaly and growth defect (2017)
  • Author(s): Anie Day Dc. Asa, Rujira Wanotayan, Mikio Shimada, Yoshihisa Matsumoto
  • Organizer: 63rd Annual International Meeting of Radiation Research Society
  • Int'l Joint Research
• [Presentation] Implication of DNA double-strand break repair in cancer, aging and development (2017)
  • Author(s): Yoshihisa Matsumoto
  • Organizer: The 12th International conference & 5th Asian Congress on Environmental Mutagens
  • Int'l Joint Research / Invited
• [Presentation] XRCC4 phosphorylation as an in situ indicator for DNA-PK activity (2017)
  • Author(s): Mukesh Kumar Sharma, Ali Reza Amiri Moghani, Mikoto Fukuchi, Shoji Imamichi, Anie Day Asa De Castro, Rujira Wanotayan, Mikio Shimada, Yoshihisa Matsumoto
  • Organizer: The 12th International conference & 5th Asian Congress on Environmental Mutagens
  • Int'l Joint Research
• [Presentation] Characterization of XRCC4 Ser260 and Ser320 phosphorylation by DNA-PK (2017)
  • Author(s): Ali Reza Amiri Moghani, Mukesh Kumar Sharma, Yoshihisa Matsumoto
  • Organizer: 33rd International Symposium of Radiation Biology Center, Kyoto University
  • Int'l Joint Research
• [Presentation] The functional analysis of DNA repair factor PNKP and susceptibility to synthetic lethality of BRCA 1/2 mutation (2017)
  • Author(s): Naoya Kase, Mikio Shimada, Hirofumi Nakano, Hiroyuki Nakamura, Yoshihisa Matsumoto
  • Organizer: 33rd International Symposium of Radiation Biology Center, Kyoto University
  • Int'l Joint Research
• [Presentation] Function of histone chaperone Anti-silencing function 1 (Asf1) in DNA damage response of human cells (2017)
  • Author(s): Akane Yamasaki, Mikio Shimada, Yoshihisa Matsumoto
  • Organizer: 33rd International Symposium of Radiation Biology Center, Kyoto University
  • Int'l Joint Research
• [Presentation] DNA二重鎖切断修復欠損細胞の低線量率放射線に対する感受性 (2017)
  • Author(s): 土屋 尚代, 島田 幹男, 小林 純也, 松本 義久
  • Organizer: 第23回癌治療増感研究会
• [Presentation] Behavior of DNA double-strand break repair-deficient cells under low dose rate irradiation (2017)
  • Author(s): Yoshihisa Matsumoto, Hisayo Tsuchiya, Mikio Shimada, Junya Kobayashi
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] DNA double-strand break repair and V(D)J recombination function of XRCC4 mutants with microcephaly and growth defect (2017)
  • Author(s): Anie Day ASA, Rujira WANOTAYAN, Mikio SHIMADA, Yoshihisa MATSUMOTO
  • Organizer: 日本放射線影響学会第60回大会
• [Presentation] DNA修復因子PNKPの機能解析とBRCA1/2変異との合成致死における感受性の検討 (2017)
  • Author(s): 加瀬 直也, 島田 幹男, 中野 洋文, 中村 浩之, 松本 義久
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] ヒト細胞のDNA損傷応答におけるヒストンシャペロンAnti-silencing function 1の機能の解析 (2017)
  • Author(s): 山崎 あかね, 島田 幹男, 松本 義久
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] XRCC4: target of phosphorylation by DNA-PK in DNA double-strand break repair. (2016)
  • Author(s): Mukesh Kumar Sharma, Shoji Imamichi, Mikoto Fukuchi, Ali Reza Amiri Moghani, Rujira Wanotayan, Mikio Shimada, Yoshihisa Matsumoto.
  • Organizer: 10th 3R International Symposium
  • Place of Presentation: Hotel Ichibata (Matsue)
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] DNA damage-induced phosphorylation of XRCC4 and its role in DNA double-strand break repair. (2016)
  • Author(s): Ali Reza Amiri Moghani, Rujira Wanotayan, Mukesh Kumar Sharma, Anie Day Asa De Castro, Mikio Shimada, Yoshihisa Matsumoto.
  • Organizer: The Fifth International Symposium on Innovative Nuclear Energy Systems
  • Place of Presentation: Tokyo Institute of Technology (Tokyo)
  • Year and Date: 2016-10-31
  • Int'l Joint Research
• [Presentation] DNA damage-induced phosphorylation of XRCC4 at multiple sites. (2016)
  • Author(s): Ali Reza Amiri Moghani, Rujira Wanotayan, Mukesh Kumar Sharma, Anie Day Asa De Castro, 島田 幹男, 松本 義久.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: JMSアステールプラザ(広島)
  • Year and Date: 2016-10-26
• [Presentation] DNA二重鎖切断修復遺伝子欠損細胞(ヒト細胞)の作製と解析. (2016)
  • Author(s): 松宮 雅典, 松本 義久, 島田 幹男.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: JMSアステールプラザ(広島)
  • Year and Date: 2016-10-26
• [Presentation] DNA double strand break repair function of XRCC4 mutants associated with microcephaly and growth defect. (2016)
  • Author(s): Anie Day Asa De Castro, 松本 義久, 島田 幹男.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: JMSアステールプラザ(広島)
  • Year and Date: 2016-10-26
• [Presentation] 低線量率放射線応答におけるDNA二重鎖切断修復タンパク質の役割. (2016)
  • Author(s): 土屋 尚代, 島田 幹男, 小林 純也, 松本 義久.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: JMSアステールプラザ(広島)
  • Year and Date: 2016-10-26
• [Presentation] 放射線と温熱の併用処理時のDNA損傷応答メカニズムの解析. (2016)
  • Author(s): 山口 基貴, 松本 義久, 島田 幹男.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: JMSアステールプラザ(広島)
  • Year and Date: 2016-10-26
• [Presentation] DNA double strand break repair function of XRCC4 mutants associated with microcephaly and growth defect. (2016)
  • Author(s): Anie Day Asa De Castro, Rujira Wanotayan, Mikio Shimada, Yoshihisa Matsumoto.
  • Organizer: 62nd Annual International Meeting Radiation Research Society
  • Place of Presentation: Big Island (Hawaii), USA
  • Year and Date: 2016-10-16
  • Int'l Joint Research
• [Presentation] DNA修復酵素PNKPのゲノム安定性維持における役割. (2016)
  • Author(s): 島田 幹男, 松本 義久.
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター(仙台)
  • Year and Date: 2016-09-25
• [Presentation] DNA二重鎖切断の認識・修復の分子機構 (2016)
  • Author(s): 松本 義久
  • Organizer: 日本薬学会 アイソトープ薬学研究会
  • Place of Presentation: 横浜ベイホテル東急(横浜)
  • Year and Date: 2016-03-27
  • Invited
• [Presentation] DNA二重鎖切断の認識と修復のタンパク質翻訳後修飾による統御 (2015)
  • Author(s): 松本 義久
  • Organizer: JAEA第1050回金曜セミナー「ストレス応答タンパク質のエピジェネティックな化学修飾と量子ビームによる立体構造変化の追跡」
  • Place of Presentation: いばらき量子ビームセンター(茨城・東海)
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] DNA依存性プロテインキナーゼ(DNA-PK)の機能と放射線感受性の予測・制御 (2015)
  • Author(s): 松本 義久
  • Organizer: BMB2015(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(神戸国際会議場他)
  • Year and Date: 2015-12-01
• [Presentation] 低線量率γ線連続照射下における非相同末端結合の重要性 (2015)
  • Author(s): 冨田 雅典, 小林 純也, 大塚 健介, 松本 義久, 内海 博司.
  • Organizer: BMB2015(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(神戸国際会議場他)
  • Year and Date: 2015-12-01
• [Presentation] Mechanisms of the repair of DNA double-strand breaks (2015)
  • Author(s): Yoshihisa Matsumoto
  • Organizer: YITP & YIPQS Workshop Biological & Medical Science based on Physics: Radiation and phyiscs, Physics on medical science, Modeling for biological system
  • Place of Presentation: Kyoto (Kyoto University Yukawa Memorial Center)
  • Year and Date: 2015-11-05
  • Int'l Joint Research / Invited
• [Presentation] Recognition and Repair of DNA Double-strand Breaks - Molecular Mechanisms and Implication in Medicine (2015)
  • Author(s): Yoshihisa Matsumoto
  • Organizer: International Workshop on Ionizing and Non-ionizing Radiation Influence on Structure and Biophysical Properties of Living Cells
  • Place of Presentation: Tsughkadzor, Armenia (Hotel Russia)
  • Year and Date: 2015-09-25
  • Int'l Joint Research / Invited
• [Remarks] 東京工業大学 科学技術創成研究院 先導原子力研究所 松本研究室
  • URL: http://www.nr.titech.ac.jp/~yoshim/
• [Remarks] 松本研究室
  • URL: http://www.nr.titech.ac.jp/~yoshim/
• [Remarks] 松本研究室
  • URL: http://www.nr.titech.ac.jp/~yoshim/matsumotos_lab/Home.html