Regulatory mechanisms of CtIP nuclease during DNA crosslink repair

• Project/Area Number: 24310042
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Kyoto University
• Principal Investigator: TAKATA Minoru 京都大学, 放射線生物研究センター, 教授 (30281728)
• Co-Investigator(Kenkyū-buntansha): UNNO Junya 京都大学, 放射線生物研究センター, 研究員 (50614153) ; KATSUKI Yoko 京都大学, 放射線生物研究センター, 研究員 (00645377)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000); Fiscal Year 2014: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2013: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2012: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: Fanconi貧血 / FANCD2 / CtIP / ユビキチン化 / DNAクロスリンク損傷 / DNA損傷修復 / ファンコニ貧血 / end resection / クロスリンク修復 / モノユビキチン化 / Fanconi anemia / DNA損傷応答 / DNA修復 / チェックポイント

Outline of Final Research Achievements

The Fanconi anemia (FA) pathway is critically involved in the maintenance of hematopoietic stem cells and the suppression of carcinogenesis. A key FA protein, FANCD2, is monoubiquitinated and accumulates in chromatin in response to DNA interstrand crosslinks (ICLs), where it coordinates DNA repair through mechanisms that are still poorly understood. Here we report that CtIP protein is a direct interactor of FANCD2. A region spanning amino acids 166 to 273 of CtIP, and monoubiquitination of FANCD2, were both essential for the FANCD2-CtIP interaction and MMC-induced CtIP foci. Remarkably, both FANCD2 and CtIP were critical for MMC-induced RPA2 hyperphosphorylation, an event that accompanies end resection of double strand breaks. Collectively, our results reveal a role of monoubiquitinated FANCD2 in end resection, anchoring CtIP to chromatin during ICL repair.

Research Products

• [Journal Article] Defective FANCI Binding by a Fanconi Anemia-Related FANCD2 Mutant. (2014)
  • Author(s): Sato K, Ishiai M, Takata M, Kurumizaka H.
  • Journal Title: PLoS One Volume: 9 Issue: 12 Pages: e114752-e114752
  • DOI: 10.1371/journal.pone.0114752
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The Trp53-Trp53inp1-Tnfrsf10b Pathway Regulates the Radiation Response of Mouse Spermatogonial Stem Cells. (2014)
  • Author(s): Kei Ishii, Masamichi Ishiai, Hiroko Morimoto, Mito Kanatsu-Shinohara, Ohtsura Niwa, Minoru Takata, and Takashi Shinohara.
  • Journal Title: Stem Cell Reports. Volume: 3 Issue: 4 Pages: 676-89
  • DOI: 10.1016/j.stemcr.2014.08.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Expression and purification of human FANCI and FANCD2 using Escherichia coli cells. (2014)
  • Author(s): Daisuke Takahashi, Koichi Sato, Mayo Shimomuki, Minoru Takata, Hitoshi Kurumizaka
  • Journal Title: Protein Expression and Purification Volume: 103 Pages: 8-15
  • DOI: 10.1016/j.pep.2014.08.012
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Modularized functions of the Fanconi anemia core complex. (2014)
  • Author(s): Huang Y, Leung JW, Lowery M, Matsushita N, Wang Y, Shen X, Huong D, Takata M, Chen J, Li L.
  • Journal Title: Cell Rep. Volume: 7 Issue: 6 Pages: 1849-57
  • DOI: 10.1016/j.celrep.2014.04.029
  • Peer Reviewed / Open Access
• [Journal Article] FANCD2 Binds CtIP and Regulates DNA-End Resection during DNA Interstrand Crosslink Repair. (2014)
  • Author(s): Unno J
  • Journal Title: Cell Rep. Volume: 7 Issue: 4 Pages: 1039-1047
  • DOI: 10.1016/j.celrep.2014.04.005
  • Peer Reviewed / Open Access
• [Journal Article] Tumor suppressor RecQL5 controls recombination induced by DNA crosslinking agents (2014)
  • Author(s): Yoshifumi Hosono, Takuya Abe, Masamichi Ishiai, M. Nurul Islam, Hiroshi Arakawa, Weidong Wang, Shunichi Takeda, Yutaka Ishii, Minoru Takata, Masayuki Seki, Takemi Enomoto
  • Journal Title: Biochimica et Biophysica Acta-Molecular Cell Research Volume: 1843 Issue: 5 Pages: 1002-1012
  • DOI: 10.1016/j.bbamcr.2014.01.005
  • Peer Reviewed / Open Access
• [Journal Article] A novel interplay between the Fanconi anemia core complex and ATR-ATRIP kinase during DNA cross-link repair (2013)
  • Author(s): J Tomida, A Itaya, T Shigechi, J Unno, E Uchida, M Ikura, Y Masuda, S Matsuda J Adachi, M Kobayashi, AR Meetei, Y Maehara, K Yamamoto K Kamiya, A Matsuura, T Matsuda. T Ikura, M Ishiai, M Takata
  • Journal Title: Nucleic Acids Research Volume: VOL. 41 Issue: 14 Pages: 6930-6941
  • DOI: 10.1093/nar/gkt467
  • Peer Reviewed
• [Journal Article] Variant ALDH2 is associated with accelerated progression of bone marrow failure in Japanese Fanconi anemia patients. (2013)
  • Author(s): Hira A, Yoshida K
  • Journal Title: Blood Volume: 122 Issue: 18 Pages: 3206-3209
  • DOI: 10.1182/blood-2013-06-507962
  • Peer Reviewed
• [Journal Article] Vandetanib is effective in EGFR-mutant lung cancer cells with PTEN deficiency. (2013)
  • Author(s): Takeda H, Takigawa N, Ohashi K, Minami D, Kataoka I, Ichihara E, Ochi N, Tanimoto M, Kiura K.
  • Journal Title: Mol Cancer Res Volume: 319 Issue: 2 Pages: 417-423
  • DOI: 10.1158/1541-7786.mcr-12-0401
  • NAID: 120005359097
  • Peer Reviewed
• [Journal Article] NBS1 directly activates ATR independently of MRE11 and TOPBP1. (2013)
  • Author(s): Kobayashi M, Hayashi N, Takata M, Yamamoto K.
  • Journal Title: Genes Cells. Volume: 18 Issue: 3 Pages: 238-46
  • DOI: 10.1111/gtc.12031
  • Peer Reviewed
• [Journal Article] DNA robustly stimulates FANCD2 monoubiquitylation in the complex with FANCI (2012)
  • Author(s): Koichi Sato. Kazue Toda, Masamichi Ishiai, Minora Takata, Hitoshi Kurumizaka
  • Journal Title: Nucleic Acids Research Volume: 40 Issue: 10 Pages: 4553-4561
  • DOI: 10.1093/nar/gks053
  • Peer Reviewed
• [Journal Article] Matched sibling donor stem cell transplantation for Fanconi anemia patients with T-cell somatic mosaicism (2012)
  • Author(s): Yabe M, Shimizu T, Morimoto T, Koike T, Takakura H, Tsukamoto H, Muroi K, Oshima K, Asami K, Takata M, Yamashita T, Kato S, Yabe H
  • Journal Title: Pediatric Transplant Volume: (印刷中) Issue: 4 Pages: 340-345
  • DOI: 10.1111/j.1399-3046.2012.01669.x
  • Peer Reviewed
• [Journal Article] Histone chaperone activity of Fanconi anemia proteins, FANCD2 and FANCI, is required for DNA crosslink repair (2012)
  • Author(s): Koichi Sato. Masamichi Ishiai, Kazue Toda, Satoshi Furukoshi, Akihisa Osakabe, Hiroaki Tachiwana, Yoshimasa Takizawa, Wataru Kagawa, Hiroyuki Kitao, Naoshi Dohmae, Chikashi Obuse, Hiroshi Kimura, Minoru Takata, Hitoshi Kurumizaka
  • Journal Title: The EMBO Journal Volume: 31 Issue: 17 Pages: 3524-3536
  • DOI: 10.1038/emboj.2012.197
  • Peer Reviewed
• [Journal Article] Mcm8 and Mcm9 form a complex that functions in homologous recombination induced by interstrand crosslinks (2012)
  • Author(s): K. Nishimura
  • Journal Title: Molecular Cell Volume: 47 Issue: 4 Pages: 511-522
  • DOI: 10.1016/j.molcel.2012.05.047
  • Peer Reviewed
• [Journal Article] A ubiquitin-binding protein, FAAP20, links RNF8-mediated ubiquitination to the Fanconi anemia DNA repair network. (2012)
  • Author(s): Yan Z, Guo R, Paramasivam M, Shen W, Ling C, Fox D 3rd, Wang Y, Oostra AB, Kuehl J, Lee DY, Takata M, Hoatlin ME, Schindler D, Joenje H, de Winter JP, Li L, Seidman MM, Wang W
  • Journal Title: Mol Cell. Volume: 47 Issue: 1 Pages: 61-75
  • DOI: 10.1016/j.molcel.2012.05.026
  • Peer Reviewed
• [Journal Article] Establishment of the DNA repair-defective mutants in DT40 cells. (2012)
  • Author(s): Ishiai M, Uchida E, Takata M.
  • Journal Title: Methods Mol Biol. Volume: 920 Pages: 39-49
  • DOI: 10.1007/978-1-61779-998-3_4
  • NAID: 120004996387
  • ISBN: 9781617799976, 9781617799983
  • Peer Reviewed
• [Presentation] 日本人ファンコニ貧血患者における新規原因遺伝子UBE2Tの同定 (2014)
  • Author(s): 平 明日香, 吉田 健一, 佐藤 浩一, 嶋本 顕, 田原 栄俊, 胡桃坂 仁志, 小川 誠司, 高田 穣, 矢部 普正, 矢部 みはる
  • Organizer: 第37回日本分子生物学会年会 ワークショップ
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] ファンコニ貧血経路とそのキータンパク質FANCD2の機能解析 (2014)
  • Author(s): 高田 穣, 勝木 陽子, 佐藤 浩一, 石合 正道, 胡桃坂 仁志
  • Organizer: 第37回日本分子生物学会年会 ワークショップ
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] UBE2T is a novel FA gene identified in Japanese Fanconi anemia patients (2014)
  • Author(s): Minoru Takata, Asuka Hira, Kenichi Yoshida, Koichi Sato, Akira Shimamoto, Hidetoshi Tahara, Hitoshi Kurumizaka, Seishi Ogawa, Hiromasa Yabe, and Miharu Yabe
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: Gotemba Kogen Hotel
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] Comprehensive analysis of Japanese Fanconi anemia (FA) patients has led to the identification of an E2 enzyme UBE2T as a novel FA gene. (2014)
  • Author(s): Minoru Takata
  • Organizer: 日米修復会議
  • Place of Presentation: エクシブ鳴門
  • Year and Date: 2014-10-29
  • Invited
• [Presentation] Infant Japanese Fanconi anemia patients with the ALDH2-AA genotype. (2014)
  • Author(s): M. Yabe, A. Hira, H, Yabe, T. Morimoto, A. Fukumura, M. Miyashita, K. Ohtsubo, K. Matsuo, M. Takata.
  • Organizer: 26th Annual Fanconi Anemia Research Fund Scientific Symposium.
  • Place of Presentation: Bethesda MA, USA
  • Year and Date: 2014-09-18 – 2014-09-21
• [Presentation] 遺伝性DNA修復異常疾患：家族性乳がんとファンコニ貧血 (2014)
  • Author(s): 高田 穣
  • Organizer: 日本家族性腫瘍学会 第１７回家族性腫瘍セミナー
  • Place of Presentation: 近畿大学・東大阪キャンパス
  • Year and Date: 2014-08-24
  • Invited
• [Presentation] ファンコニ貧血症とDNA修復メカニズム (2014)
  • Author(s): 髙田 穣
  • Organizer: 第３６回日本光医学・光生物学会シンポジウム
  • Place of Presentation: 大阪大学銀杏会館
  • Year and Date: 2014-07-25
  • Invited
• [Presentation] FANCD2 in chromatin anchors CtIP and regulates DNA end resection during crosslink repair. (2013)
  • Author(s): J. Unno, A. Itaya, M. Taoka, K. Sato, J. Tomida, W. Sakai, T. Ikura, T. Isobe, H. Kurumizaka, M. Takata
  • Organizer: 25th Annual Fanconi anemia research fund Scientific Symposium
  • Place of Presentation: Houston, Texas, USA
• [Presentation] プログレス教育講演 「ファンコニ貧血とＤＮＡクロスリンク修復の分子機構 ー最近の進歩ー」 (2013)
  • Author(s): 高田 穣
  • Organizer: 日本血液学会
  • Place of Presentation: 札幌市
  • Invited
• [Presentation] Crosstalk signaling between checkpoint and the Fanconi anemia DNA repair pathway (2012)
  • Author(s): Junya Unno, Junya Tomida, Tomoko Shigechi, Akiko l taya, and Minoru Takata
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌・ホテルさっぽろ芸文館・札幌市教育文化会館(北海道)
• [Presentation] Genetic interplay between the Fanconi anemia pathway and aldehyde metabolism in humans (2012)
  • Author(s): Minoru Takata, Asuka Hira, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: The 8th 3R Symposium
  • Place of Presentation: 淡路夢舞台国際会議場(兵庫県)
• [Presentation] Genetic interplay between the Fanconi anemia pathway and aldehyde metabolism in humans (2012)
  • Author(s): Minoru Takata, Asuka Hiral, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: 28th RBC-NIRS INTERNATIONAL SYMPOSIUM
  • Place of Presentation: コープイン京都(京都府)
• [Presentation] Molecular pathogenesis of Fanconi anemia (2012)
  • Author(s): Minoru Takata
  • Organizer: 第54回日本小児血液・がん学会学術集会
  • Place of Presentation: パシフィコ横浜(神奈川県)
• [Presentation] Fanconi貧血とゲノム不安定性
  • Author(s): 高田 穣
  • Organizer: 第13回血液フォーラム21 －造血不全－
  • Place of Presentation: 東京フォーラム
  • Invited
• [Remarks] 京都大学 放射線生物研究センター
  • URL: http://www.rbc.kyoto-u.ac.jp
• [Remarks]
  • URL: http://house.rbc.kyoto-u.ac.jp/Late_Effect/Japanese_site/Welcom.html