Construction and application of linear ubiquitin-specific molecular probes for anti-cancer drug screening

• Project/Area Number: 25670137
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: Gunma University
• Principal Investigator: TOKUNAGA Fuminori 群馬大学, 生体調節研究所, 教授 (00212069)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 酵素 / タンパク質 / 細胞・ 組織 / バイオテクノロジー / 生体機能利用 / シグナル伝達 / 免疫学

Outline of Final Research Achievements

NF-κB signal plays a critical role on immune and inflammatory responses induced by various stimuli such as bacterial and viral infections. We identified a novel enzyme (named LUBAC) that generates head-to-tail-linked linear polyubiquitin chain, and LUBAC is critical for NF-κB regulation. Furthermore, we identified a protein domain (A20 ZF7), which exhibits linear ubiquitin-specific binding. We here employed the domain to detect intracellular linear ubiquitin and screening for linear ubiquitin-binding chemicals.

Research Products

• [Journal Article] Structures of CYLD USP with Met1- or Lys63-linked diubiquitin reveal mechanisms for dual specificity (2015)
  • Author(s): Sato, Y., Goto, E., Shibata, Y., Kubota, Y., Yamagata, A., Goto-Ito, S., Kubota, K., Inoue, J., Takekawa, M., Tokunaga, F. and Fukai, S.
  • Journal Title: Nat. Struct. Mol. Biol. Volume: 22 Issue: 3 Pages: 222-229
  • DOI: 10.1038/nsmb.2970
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A novel autosomal recessive mutation of DSG4 causes monilethrix through the ER stress response. (2015)
  • Author(s): Madoka Kato, Akira Shimizu, Yoko Yokoyama, Kyoichi Kaira, Yutaka Shimomura, Akemi Ishida-Yamamoto, Kiyoko Kamei, Fuminori Tokunaga, and Osamu Ishikawa
  • Journal Title: J. Invest. Dermatol. Volume: 135 Issue: 5 Pages: 1253-1260
  • DOI: 10.1038/jid.2015.12
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 8番目のユビキチン鎖、直鎖状ユビキチン鎖の機能と疾患への関わり： 炎症、免疫応答に重要なNF-κB経路の新しい制御機構 (2014)
  • Author(s): 徳永文稔
  • Journal Title: 化学と生物 Volume: 52 Pages: 716-718
• [Journal Article] Structural and functional analyses of DNA-sensing and immune activation by human cGAS (2013)
  • Author(s): Kazuki Kato, Ryohei Ishii, Eiji Goto, Ryuichiro Ishitani, Fuminori Tokunaga, and Osamu Nureki
  • Journal Title: PLoS One Volume: 8 Issue: 10 Pages: e76983-e76983
  • DOI: 10.1371/journal.pone.0076983
  • Peer Reviewed
• [Journal Article] Recruitment of the autophagic machinery to endosomes during infection is mediated by ubiquitin (2013)
  • Author(s): Naonobu Fujita et al.
  • Journal Title: J. Cell Biol. Volume: 203 Issue: 1 Pages: 115-128
  • DOI: 10.1083/jcb.201304188
  • Peer Reviewed / Open Access
• [Journal Article] Linear ubiquitination-mediated NF-κB regulation and its related disorders (2013)
  • Author(s): Fuminori Tokunaga
  • Journal Title: J. Biochem. Volume: 154 Issue: 4 Pages: 313-323
  • DOI: 10.1093/jb/mvt079
  • Peer Reviewed
• [Journal Article] 直鎖状ユビキチン化を介したNF-κB制御機構と疾患 (2013)
  • Author(s): 徳永文稔
  • Journal Title: 生化学 Volume: 85 Pages: 414-422
  • NAID: 10031184261
• [Presentation] Linear ubiquitination-mediated NF-κB regulation. (2015)
  • Author(s): Fuminori Tokunaga
  • Organizer: 2nd International Symposium on Protein Modifications in Pathogenic Dysregulation of Signaling.
  • Place of Presentation: 東京大学医科学研究所（東京）
  • Year and Date: 2015-01-23
  • Invited
• [Presentation] Optineurinの直鎖状ユビキチン結合を介したNF-κB制御と疾患との関連 (2014)
  • Author(s): 及川大輔、石井亮平、中澤世識、石谷隆一郎、濡木理、徳永文稔
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2014-11-26
• [Presentation] RISPR/Cas9法を用いたLUBAC欠損細胞の作製とNF-κB活性制御への影響 (2014)
  • Author(s): 後藤栄治、徳永文稔
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2014-11-26
• [Presentation] OptineurinのNF-κBシグナル制御の細胞機構と疾患 (2014)
  • Author(s): 及川大輔、石井亮平、中澤世識、石谷隆一郎、濡木理、徳永文稔
  • Organizer: 第９回臨床外レス応答学会
  • Place of Presentation: 岡山大学（岡山市）
  • Year and Date: 2014-11-01
• [Presentation] 癌抑制遺伝子STK11による細胞死制御機構 (2014)
  • Author(s): 野口拓也、熊澤琢也、徳永文稔
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都国際会館（京都市）
  • Year and Date: 2014-10-17
• [Presentation] 直鎖状ユビキチン鎖生成を介したNF-κBシグナル制御の分子基盤 (2014)
  • Author(s): 徳永文稔
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都国際会館（京都市）
  • Year and Date: 2014-10-16
  • Invited
• [Presentation] 新規LUBACシグナル複合体構成因子の探索と細胞機能解析 (2014)
  • Author(s): 及川大輔、中澤世識、徳永文稔
  • Organizer: 第66回日本細胞生物学会
  • Place of Presentation: 奈良県新公会堂（奈良市）
  • Year and Date: 2014-06-30
• [Presentation] 直鎖状ポリユビキチン修飾を介したNF-κB制御のB細胞リンパ腫発症における役割
  • Author(s): 徳永文稔
  • Organizer: 日本薬学会第134年会
  • Place of Presentation: 熊本
  • Invited
• [Presentation] 直鎖状ユビキチン鎖生成を介したNF-κBシグナル制御と疾患
  • Author(s): 徳永文稔
  • Organizer: 東京工業大学バイオサイエンスシンポジウム
  • Place of Presentation: 横浜
  • Invited
• [Presentation] ユビキチン修飾による細胞機能制御と疾患
  • Author(s): 徳永文稔
  • Organizer: 第60回北関東医学会総会
  • Place of Presentation: 前橋
  • Invited
• [Presentation] Linear ubiquitination-mediated NF-κB regulation mechanism and its related disorders
  • Author(s): Fuminori Tokunaga
  • Organizer: プロテイン・アイランド・松山(PIM)国際シンポジウム2013
  • Place of Presentation: 松山
  • Invited
• [Presentation] 直鎖状ポリユビキチン化を介したNF-κB制御と疾患
  • Author(s): 徳永文稔
  • Organizer: 第86回日本生化学会 シンポジウム
  • Place of Presentation: 横浜
  • Invited
• [Presentation] 直鎖状ユビキチン化を介するNF-κB制御に関わる脱ユビキチン化酵素と疾患
  • Author(s): 徳永文稔、西増弘志、石谷隆一郎、濡木理
  • Organizer: 病態プロテアーゼ学会
  • Place of Presentation: 大阪
• [Presentation] Down-regulation mechanism for LUBAC-induced NF-κB pathway
  • Author(s): Fuminori Tokunaga
  • Organizer: The 35th Naito Conference: The ubiquitin-proteasome system: From basic mechanisms to pathophysiological roles
  • Place of Presentation: Sapporo
• [Remarks] 群馬大学生体調節研究所分子細胞制御分野ホームページ
  • URL: http://www.imcr.gunma-u.ac.jp/lab/molcb/
• [Remarks] 群馬大学生体調節研究所ホームページ
  • URL: http://www.imcr.gunma-u.ac.jp/
• [Remarks] 群馬大学生体調節研究所分子細胞制御分野ホームページ
  • URL: http://www.imcr.gunma-u.ac.jp/lab/molcb/
• [Remarks] 群馬大学生体調節研究所ホームページ
  • URL: http://www.imcr.gunma-u.ac.jp/