Construction of polyubiquitin chain specific molecular probes and their applications to histopathologic examinations.

• Project/Area Number: 23657075
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• 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): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ユビキチン / 炎症 / NF-κB / 細胞内シグナル伝達 / 分子プローブ / シグナル伝達 / 細胞・組織 / 酵素 / タンパク質

Research Abstract

The ubiquitin system functions as a pivotal post-transcriptional modification in numerous cellular functions, including protein degradation and signal transduction. We identified that LUBAC (linear ubiquitin chain assembly complex) generates a novel type of Met1-linked linear polyubiquitin chain and specifically regulates the NF-κB pathway, which is a central signaling pathway for inflammatory and immune responses. In this study, we show that A20, a deubiquitinase, suppresses NF-κB activation by binding linear polyubiquitin via the C-terminal seventh zinc finger (ZF7), and we determined the crystal structures of A20 ZF7 in complex with linear diubiquitin. Importantly, we found that a tandem conjugate of three ZF7s is available as a high affinity molecular probe to detect linear poluubiquitin chain.

Research Products

• [Journal Article] Specific recognition of linear polyubiquitin by A20 zinc finger 7 is involved in NF-κB regulation (2012)
  • Author(s): Fuminori Tokunaga, Hiroshi Nishimasu, Ryuichiro Ishitani, Eiji Goto, Takuya Noguchi, Kazuhiro Mio, Kiyoko Kamei, Averil Ma, Kazuhiro Iwai, and Osamu Nureki
  • Journal Title: EMBO J Volume: 31 Pages: 3856-3870
  • Peer Reviewed
• [Journal Article] Analysis of NF-κB essential modulator (NEMO) binding to linear and lysine-linked ubiquitin chains and its role in the activation of NF-κB (2012)
  • Author(s): Tobias Kensche, Fuminori Tokunaga, Fumiyo Ikeda, Eiji Goto, Kazuhiro Iwai, and Ivan Dikic
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 23626-23634
  • Peer Reviewed
• [Journal Article] Non-canonical UBA-UBL interaction mediates formation of linear ubiquitin chain assembly complex (2012)
  • Author(s): Hirokazu Yagi, Kazuhiro Ishimoto, Takeshi Hiromoto, Hiroaki Fujita, Tsunehiro Mizushima, Yoshinori Uekusa, Maho Yagi-Utsumi, Eiji Kurimoto, Masanori Noda, Susumu Uchiyama, Fuminori Tokunaga, Kazuhiro Iwai, and Koichi Kato
  • Journal Title: EMBO Rep Volume: 13 Pages: 462-468
  • Peer Reviewed
• [Journal Article] Backbone and side chain 1H, 13C, and 15N assignments of ubiquitin-like domain of human HOIL-1L, an essential component of linear ubiquitin chain assembly complex (2012)
  • Author(s): Yoshinori Uekusa, Syunsuke Miura, Hiroaki Sasakawa, Eiji Kurimoto, Eri Sakata, Serve Oliver, Hirokazu Yagi, Fuminori Tokunaga,Kazuhiro Iwai, and Koichi Kato
  • Journal Title: Biomol. NMR Assign. Volume: 6 Pages: 177-180
  • Peer Reviewed
• [Journal Article] Activation of nuclear factor-kappa B by linear ubiquitin chain assembly complex contributes to lung metastasis of osteosarcoma cells (2012)
  • Author(s): Masato Tomonaga, Nobuyuki Hashimoto, Fuminori Tokunaga, Megumi Onishi, Akira Myoui, Hideki Yoshikawa, and Kazuhiro Iwai
  • Journal Title: Int. J.Oncol Volume: 40 Pages: 409-417
  • Peer Reviewed
• [Journal Article] A novel NF-κB regulatory mechanism for inflammatory and immune responses by the LUBAC ubiquitin ligase complex (2012)
  • Author(s): Fuminori Tokunaga and Kazuhiro Iwai: Linear ubiquitination
  • Journal Title: Endocrine J. Volume: 59 Pages: 641-652
  • Peer Reviewed
• [Journal Article] LUBAC, a novel ubiquitin ligase for linear ubiquitination, is crucial for inflammation and immune responses (2012)
  • Author(s): Fuminori Tokunaga and Kazuhiro Iwai
  • Journal Title: Microbes Infect. Volume: 14 Pages: 563-572
  • Peer Reviewed
• [Journal Article] Specific recognition of linear polyubiquitin by A20 zinc finger 7 is involved in NF-κB regulation (2012)
  • Author(s): Tokunaga F, Nishimasu H, Ishitani R, Goto E, Noguchi T, Mio K, Kamei K, Ma A, Iwai K, Nureki O
  • Journal Title: EMBO J. Volume: 31 Issue: 19 Pages: 3856-3870
  • DOI: 10.1038/emboj.2012.241
  • Peer Reviewed
• [Journal Article] Analysis of nuclear factor-κB (NF-κB) essential modulator (NEMO) binding to linear and lysine-linked ubiquitin chains and its role in the activation of NF-κB. (2012)
  • Author(s): Kensche, T.
  • Journal Title: J. Biol. Chem. Volume: 287 Issue: 28 Pages: 23626-23634
  • DOI: 10.1074/jbc.m112.347195
  • Peer Reviewed
• [Journal Article] A non-canonical UBA-UBL interaction forms the linear-ubiquitin-chain assembly complex. (2012)
  • Author(s): H.Yagi, K.Ishimoto, T.Hiromoto, H.Fujita, T.Mizushima, Y.Uekusa, M.Yagi-Utsumi, E.Kurimoto, M.Noda, S.Uchiyama, F.Tokunaga, K. Iwai, and K.Kato
  • Journal Title: EMBO Rep. Volume: 13 Issue: 5 Pages: 462-468
  • DOI: 10.1038/embor.2012.24
  • Peer Reviewed
• [Journal Article] A novel NF-κB regulatory mechanism for inflammatory and immune responses by the LUBAC ubiquitin ligase complex. (2012)
  • Author(s): Tokunaga, F.
  • Journal Title: Endocr. J. Volume: 59 Pages: 641-652
  • Peer Reviewed
• [Journal Article] LUBAC, a novel ubiquitin ligase for linear ubiquitination, is crucial for inflammation and immune responses (2012)
  • Author(s): Tokunaga, F., et al.
  • Journal Title: Microbes Infect. Volume: (in press) Issue: 7-8 Pages: 563-572
  • DOI: 10.1016/j.micinf.2012.01.011
  • Peer Reviewed
• [Journal Article] Backbone and side chain ^1H, ^<13>C, and ^<15>N assignments of the ubiquitin-like domain of human HOIL-1L, an essential component of linear ubiquitin chain assembly complex (2012)
  • Author(s): Y.Uekusa, S.Mimura, H.Sasakawa, E.Kurimoto, E.Sakata, S.Olivier, H.Yagi, F.Tokunaga, K.Iwai, K.Kato
  • Journal Title: Biomol.NMR Assign. Volume: (in press) Issue: 2 Pages: 177-180
  • DOI: 10.1007/s12104-011-9350-1
  • Peer Reviewed
• [Journal Article] Activation of nuclear factor-kappa B by linear ubiquitin chain assembly complex contributes to lung metastasis of osteosarcoma cells (2012)
  • Author(s): Tomonoga, M., et al.
  • Journal Title: Int.J.Oncol. Volume: 40 Pages: 409-417
  • DOI: 10.3892/ijo.2011.1209
  • Peer Reviewed
• [Journal Article] SHARPIN is a component of the NF-κB activating linear ubiquitin assembly complex (2011)
  • Author(s): Fuminori Tokunaga, Tomoko Nakagawa, Masaki Nakahara, Yasushi Saeki, Masami Taniguchi, Shin-ichi Sakata, Keiji Tanaka, Hiroyasu Nakano, and Kazuhiro Iwai
  • Journal Title: Nature Volume: 471 Pages: 633-636
  • Peer Reviewed
• [Journal Article] SHARPIN forms a linear ubiquitin ligase complex regulating NF-κB activity and apoptosis (2011)
  • Author(s): Fumiyo Ikeda, Yonathan Lissanu Deribe, Sigrid S. Skanland, Benjamin Stieglitz, Caroline Grabbe, Sjoerd van WijK, Mirita Franz-Wachtel, Panchali Goswami, Vanja Nagy, Janos Terzic, Fuminori Tokunaga, Ariadne Androulidaki, Tomoko Nakagawa, Manolis Pasparakis, Kazuhiro Iwai, John P. Sundberg, Liliana Schaefer, Boris Macek, Katrin Rittinger, and Ivan Dikic
  • Journal Title: Nature Volume: 471 Pages: 637-641
  • Peer Reviewed
• [Journal Article] Linear ubiquitin assembly complex negativelyregulates RIG-I- and TRIM25-mediated type I interferon induction (2011)
  • Author(s): Kyung-Soo Inn, Michaela U. Gack, Fuminori Tokunaga, Mude Shi, Lai-Yee Wong, Kazuhiro Iwai, and Jae U. Jung
  • Journal Title: Mol. Cell Volume: 41 Pages: 354-365
  • Peer Reviewed
• [Journal Article] Sharpin is a component of the NF-κB activating linear ubiquitin chain assembly complex (2011)
  • Author(s): Tokunaga, F., Nakagawa, T., Nakahara, M., Saeki, Y., Taniguchi, M., Sakata, S., Tanaka, K., Nakano, H., Iwai, I.
  • Journal Title: Nature Volume: 471 Issue: 7340 Pages: 633-636
  • DOI: 10.1038/nature09815
  • Peer Reviewed
• [Journal Article] SHARPIN forms a linear ubiquitin ligase complex regulating NF-κB activity and apoptosis (2011)
  • Author(s): Ikeda, F., et al.
  • Journal Title: Nature Volume: 471 Issue: 7340 Pages: 637-641
  • DOI: 10.1038/nature09814
  • Peer Reviewed
• [Presentation] Linear ubiquitin binding via A20 zinc finger 7 is involved in the canonical NF-B regulation (2013)
  • Author(s): Fuminori Tokunaga, Hiroshi Nishimasu, Ryuichiro Ishitani, and Osamu Nureki
  • Organizer: Frontiers in the immunology and inflammation: From molecules to disease.
  • Place of Presentation: Tokyo
  • Year and Date: 2013-02-12
• [Presentation] Activation and regulation mechanisms of the linear ubiquitination-mediated NF-κB pathway (2013)
  • Author(s): Fuminori Tokunaga
  • Organizer: 1st International Symposium on Protein Modification in Pathogenic Dysregulation of Signaling
  • Place of Presentation: Tokyo
  • Year and Date: 2013-02-01
• [Presentation] 直鎖状ユビキチン生成を介したNF-B制御とB細胞リンパ腫との連関 (2013)
  • Author(s): 徳永文稔
  • Organizer: がん研究分野の特性を踏まえた支援活動公開シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2013-01-30
• [Presentation] Linear ubiquitin binding via A20 zinc finger 7 is involved in the canonical NF-κB regulation. (2013)
  • Author(s): Tokunaga, F.
  • Organizer: Frontiers in the immunology and inflammation: From molecules to disease.
  • Place of Presentation: Tokyo、学術総合センター 一橋講堂
• [Presentation] Activation and regulation mechanisms of the linear ubiquitination-mediated NF-κB pathway. (2013)
  • Author(s): Tokunaga, F.
  • Organizer: 1st International Symposium on Protein Modification in Pathogenic Dysregulation of Signaling.
  • Place of Presentation: Tokyo、東大医科研
  • Invited
• [Presentation] A20の直鎖状ポリユビキチン鎖特異的結合を介したNF-B制御 (2012)
  • Author(s): 徳永文稔、西増弘志、石谷隆一郎、後藤栄治、野口拓也、岩井一宏、濡木理
  • Organizer: 第85回日本生化学会シンポジウム:ベールを脱いだユビキチン系の新機能
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-16
• [Presentation] 直鎖状ユビキチン形成とNF-Bシグナル伝達 (2012)
  • Author(s): 徳永文稔
  • Organizer: 応用数理学会数理医学研究部会オーガナイズドセッション
  • Place of Presentation: 稚内
  • Year and Date: 2012-08-30
• [Presentation] Specific recognition of linear ubiquitin by A20 zinc finger 7 is involved in NF-κB regulation (2012)
  • Author(s): Fuminori Tokunaga
  • Organizer: Keystone Symposium
  • Place of Presentation: Whistler, Canada
  • Year and Date: 2012-03-21
• [Presentation] 直鎖状ポリユビキチン鎖:NF-κB制御に関わる新たな修飾機構と疾患 (2012)
  • Author(s): 徳永文稔
  • Organizer: 第9回Osteoimmunology Forum
  • Place of Presentation: 東京
  • Year and Date: 2012-02-04
• [Presentation] A20の直鎖状ポリユビキチン鎖特異的結合を介したNF-κB制御. (2012)
  • Author(s): 徳永文稔
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡、福岡国際会議場
  • Invited
• [Presentation] 直鎖状ユビキチン形成とNF-κBシグナル伝達. (2012)
  • Author(s): 徳永文稔
  • Organizer: 応用数理学会
  • Place of Presentation: 稚内、全日空ホテル
  • Invited
• [Presentation] LUBACによる直鎖状ポリユビキチン鎖形成を介したNF-κB制御と疾患 (2011)
  • Author(s): 徳永文稔、中原匡咲、岩井一宏
  • Organizer: 第84回日本生化学会シンポジウム:拡大するユビキチンバイオロジー
  • Place of Presentation: 京都
  • Year and Date: 2011-09-23
• [Presentation] LUBACによる直鎖状ポリユビキチン鎖形成を介したNF-κB制御と疾患 (2011)
  • Author(s): 徳永文稔
  • Organizer: 第84回 日本生化学会大会シンポジウム(招待講演)
  • Place of Presentation: 京都
• [Book] NF-B経路の多様なポリユビキチン鎖による制御と疾患 (2012)
  • Author(s): 徳永文稔
  • Publisher: 実験医学増刊号
• [Book] シグナル伝達研究最前線2012 (2012)
  • Author(s): 井上純一郎
  • Total Pages: 222
  • Publisher: 羊土社
• [Book] シグナル伝達研究最前線2012 (2012)
  • Author(s): 徳永文稔
  • Total Pages: 222
  • Publisher: 羊土社
• [Book] 新規直鎖状ポリユビキチン鎖生成によるNF-Bシグナル制御 (2011)
  • Author(s): 徳永文稔、岩井一宏
  • Publisher: 実験医学
• [Remarks] 群馬大学生体調節研究所分子細胞制御分
  • URL: http://www.imcr.gunma-u.ac.jp/lab/molcb/
• [Remarks] 群馬大学 生体調節研究所 分子細胞制御分野HP
  • URL: http://www.imcr.gunma-u.ac.jp/lab/molcb/
• [Remarks] 新学術領域研究 「修飾シグナル病」HP
  • URL: http://shushoku-signal.com/