プロテアソームを基軸としたタンパク質分解系の包括的研究

2013 Fiscal Year Annual Research Report

• Project/Area Number: 21000012
• Research Institution: 公益財団法人東京都医学総合研究所
• Principal Investigator: 田中 啓二 公益財団法人東京都医学総合研究所, 生体分子先端研究分野, 所長 (10108871)
• Co-Investigator(Kenkyū-buntansha): 村田 茂穂 東京大学, 大学院薬学系研究科, 教授 (20344070)
• Project Period (FY): 2009-05-08 – 2014-03-31
• Keywords: 蛋白質 / バイオテクノロジー / 酵素 / 脳神経疾患 / 免疫学

Research Abstract

プロテアソームを基軸としたタンパク質分解系の包括的研究
1, プロテアソームの動態研究
我々は26Sプロテアソームを構成するCP(触媒粒子)とRP(調節粒子)の形成に必須の特異的な分子集合シャペロンを8種発見、それら殆ど全ての立体構造解明に成功し、プロテアソームの形成機構を分子レベルで解明した。蛍光相関分光法(FCS/FCCS : 蛍光のゆらぎを高感度に測定し、溶液中や細胞内の絶対濃度や大きさ、形状などを決定する方法)により、細胞質と核のプロテアソームの濃度が各々200nM、1mMであることを測定することに成功した。またプロテアソームが細胞質で形成された後、核に輸送されることを見出した。
2, ユビキチンの研究
我々は膜電位の低下した不良ミトコンドリアが常染色体劣性若年性パーキンソン病原因遺伝子PINK1(タンパク質リン酸化酵素)とParkin(ユビキチン連結酵素)によって識別された後、プロテアソームやオートファジーによって浄化されることを見出した。このとき、PINK1が自己リン酸化によって活性型に転換されること、そしてParkinがPINK1依存性のParkinとユビキチンのリン酸化とによって活性化される新機構を発見した。また、TR-TUBEを考案して細胞内に存在するポリユビキチン鎖の長さを決定する方法、さらにUb-catcherを作製して細胞内に存在するポリユビキチン鎖化基質を同定する方法を開発した。
3, オートファジーの研究
我々はすでに肝臓特異的オートファジー欠損マウスを長期間飼育すると肝炎・肝硬変を経て、最終的に100%の確率で肝細胞ガンが発生することを報告したが、このときオートファジー選択的基質p62がと蓄積し、p62が直接に酸化ストレスマスター転写因子Nrf2を活性化すること、そしてp62がmTORCによってリン酸化されることによってNrf2に親和性を著しく増強させることを発見した。

Research Products

• [Journal Article] Structural basis for proteasome formation controlled by an assembly chaperone Nas2. (2014)
  • Author(s): Satoh, T., Saeki, Y., Hiromoto, T., Wang, Y-H., Uekusa, Y., Yagi, H., Yoshihara, H., Yagi-Utsumi, M., Mizushima, T., Tanaka, K., and Kato, K.
  • Journal Title: Structure Volume: 22 Pages: 731-743
  • DOI: 10.1016/j.str.2014.02.014
  • Peer Reviewed
• [Journal Article] Ubiquitin is phosphorylated by PINK1 to activate parkin. (2014)
  • Author(s): Koyano, F., Okatsu, K., Kosako, H., Tamura, Y., Go, E., Kimura, M., Kimura, Y., Tsuchiya, H., Yoshihara, H., Hirokawa, T., Endo, T., Fon. E. A., Trempe, J. F., Saeki, Y., Tanaka, K., and Matsuda, N.
  • Journal Title: Nature Volume: 510(in press) Pages: 162-166
  • DOI: 10.1038/nature13392
  • Peer Reviewed
• [Journal Article] Quantitative live-cell imaging reveals spatiotemporal dynamics and cytoplasmic assembly of the 26S proteasome (2014)
  • Author(s): Pack, Chan-Gi., Yukii, H., Toh-e, A., Kudo, T., Tsuchiya, H., Kaiho, A., Sakata, E., Murata, S., Yokosawa, H., Sako, Y., Baumeister, W., Tanaka, K., and Saeki, Y
  • Journal Title: Nat Commun. Volume: 5
  • DOI: 10.1038/ncornms4996
  • Peer Reviewed
• [Journal Article] Dissection of the role of p62/Sqstml in activation of Nrf2 during xenophagy. (2014)
  • Author(s): Ishimura, R., Tanaka, K., and Komatsu, M.
  • Journal Title: FEBS Lett. Volume: 588 Pages: 822-828
  • DOI: 10.1016/j.febslet.2014.01.045.
  • Peer Reviewed
• [Journal Article] Involvement of Bag6 and the TRC pathway in proteasome assembly. (2013)
  • Author(s): Akcahane, T., Sahara, K., Yashiroda, H., Tanaka, K., and Murata, S
  • Journal Title: Nat Commun. Volume: 4 Pages: 2234
  • DOI: 10.1038/ncomms3234.
  • Peer Reviewed
• [Journal Article] Parkin catalyzed ubiquitin-ester transfer is triggered by PINK1-dependent phosphorylation. (2013)
  • Author(s): Iguchi, M., Kujuro, Y., Okatsu, K., Koyano, F., Kosako, H., Kimura, M., Suzuki, N., Uchiyama, S., Tanaka, K., and Matsuda, N.
  • Journal Title: J Biol Chem. Volume: 288 Pages: 22019-22032
  • DOI: 10.1074/jbc.M113.467530.
  • Peer Reviewed
• [Journal Article] The parallel reaction monitoring method contributes to highly sensitive polyubiquitin chain quantification. (2013)
  • Author(s): Tsuchiya, H., Tanaka, K., and Saeki, Y.
  • Journal Title: Biochem Biophys Res Commun. Volume: 436 Pages: 223-229
  • DOI: 10.1016/j.bbrc.2013.05.080.
  • Peer Reviewed
• [Journal Article] Cytoplasmic proteasomes are not indispensable for cell growth in Saccharomyces cerevisiae. (2013)
  • Author(s): Tsuchiya, H., Arai, N., Tanaka, K., and Saeki, Y.
  • Journal Title: Biochem Biophys Res Commun Volume: 436 Pages: 372-376
  • DOI: 10.1016/j.bbrc.2013.05.105.
  • Peer Reviewed
• [Journal Article] The principal PINK1 and Parkin cellular events triggered in response to dissipation of mitochondrial membrane potential occur in primary neurons. (2013)
  • Author(s): Koyano, F., Okatsu, K., Ishigaki, S., Fujioka, Y., Kimura, M., Sobue, G., Tanaka, K. and Matsuda, N
  • Journal Title: Genes Cells Volume: 18 Pages: 672-681
  • DOI: 10.1111/gtc.12066.
  • Peer Reviewed
• [Journal Article] A multi-purpose fusion tag derived from an unstructured and hyper-acidic region of the amyloid precursor protein. (2013)
  • Author(s): Sangawa, T., Tabata, S., Suzuki, K., Saeki, Y., Tanaka, K., and Takagi, J.
  • Journal Title: Protein Science Volume: 22 Pages: 840-850
  • DOI: 10.1002/pro.2254.
  • Peer Reviewed
• [Journal Article] Aire-expressing thymic medullary epithelial cells originate from β5t-expressing progenitor cells. (2013)
  • Author(s): Ohigashi, I., Zuklys, S., Sakata, M., Mayer, GE., Zhanybekova. S., Murata, S., Tanaka, K., Hollander, G., and Takahama, Y.
  • Journal Title: Proc Natl Acad Sci USA Volume: 110 Pages: 9885-9890
  • DOI: 10.1073/pnas.1301799110.
  • Peer Reviewed
• [Journal Article] Spatial arrangement and functional role of α subunits of proteasome activator PA28 in hetero-oligomeric form. (2013)
  • Author(s): Sugiyama, M., Sahashi, H., Kurimoto, E., Takata, S., Yagi, H., Kanai, K., Sakata, E., Minami, Y., Tanaka, K., and Kato K.
  • Journal Title: Biochem Biophys Res Commun. Volume: 432 Pages: 141-145
  • DOI: 10.1016/j.bbrc.2013.01.071.
  • Peer Reviewed
• [Journal Article] A dimeric PINK1-containing complex on depolarized mitochondria stimulates Parkin recruitment. (2013)
  • Author(s): Okatsu, K., Uno, M., Koyano, F., Go, E., Kimura, M., Oka, T., Tanaka, K., and Matsuda, N.
  • Journal Title: J. Biol. Chem. Volume: 288 Pages: 36372-36384
  • DOI: 10.1074/jbc.M113.509653.
  • Peer Reviewed
• [Journal Article] Endogenous nitrated nucleotide is a key mediator of autophagy and innate defense against bacteria. (2013)
  • Author(s): Ito, C., Saito, Y., Nozawa, T., Fujii, S., Inoue, H., Matsunaga, T., Khan, S., Aikawa, C., Takahashi, E., Sawa, T., Komatsu, M., Tanaka, K., Akaike, T., Nakagawa, I., and Arimoto, H
  • Journal Title: Mol Cell Volume: 52 Pages: 794-804
  • DOI: 10.1016/j.molcel.2013.10.024.
  • Peer Reviewed
• [Journal Article] Different dynamic movements of wild-type and pathogenic VCPs and their cofactors to damaged mitochondria in a Parkin-mediated mitochondrial quality control system. (2013)
  • Author(s): Kimura, Y., Fukushi, J., Hori, S., Matsuda, N., Okatsu, K., Kakiyama, Y., Kawawaki, J., Kakizuka, A., and Tanaka, K.
  • Journal Title: Genes Cells Volume: 18 Pages: 1131-1143
  • DOI: 10.1111/gtc.12103.
  • Peer Reviewed
• [Journal Article] Defective immune responses in mice lacking LUBAC-mediated linear ubiquitination in B cells (2013)
  • Author(s): Sasaki, Y., Sano, S., Nakahara, M., Murata, S., Kometani, K., Aiba, Y., Sakamoto, S., Watanabe, Y., Tanaka, K., Kurosaki, T., and lwai, K.
  • Journal Title: EMBO J. Volume: 32 Pages: 2463-2476
  • DOI: 10.1038/emboj.2013.184.
  • Peer Reviewed
• [Journal Article] Phosphorylation of p62 activates the Keap1-Nrf2 pathway during selective autophagy. (2013)
  • Author(s): Ichimura, Y., Waguri, S., Sou, Y., Kageyama, S., Hasegawa, J., Ishimura, R., Saito, T., Yang, Y., Kouno, T., Fukutomi, T., Hoshii, T., Atsushi, H., Takagi, K., Mizushima, T., Motohashi, H., Lee, M-S., Yoshimori, T., Tanaka, K., Yamamoto, M., and Komatsu, K.
  • Journal Title: Mol Cell Volume: 51 Pages: 618-631
  • DOI: 10.1016/j.molcel.2013.08.003.
  • Peer Reviewed
• [Journal Article] The proteasome : from basic mechanisms to emerging roles. (2013)
  • Author(s): Tanaka, K.
  • Journal Title: Keio J Med. Volume: 62 Pages: 1-12
  • DOI: 10.2302/kjm.2012-0006.RE
  • Peer Reviewed
• [Journal Article] Proteostasis and neurodegeneration : the roles of proteasomal degradation and autophagy. (2013)
  • Author(s): Tanaka, K. and Matsuda, N.
  • Journal Title: Biochim. Biophys. Acta (Molecular Cell Research) Volume: 1843 Pages: 197-204
  • DOI: 10.1016/j.bbamcr.2013.03.012.
  • Peer Reviewed
• [Presentation] The vertebrate-specific thymoproteasome governs positive selection in the development of CD8 T lymphocytes (2014)
  • Author(s): Keiji Tanaka
  • Organizer: The Zomes VIII Conference, Biology of the Zomes-Proteasome, COP9 Signalosome and associated processes.
  • Place of Presentation: Xiamen, China(発表確定)
  • Year and Date: 20141118-21
  • Invited
• [Presentation] Thymoproteasomes play an essential role for the formation of immunocompetent repertoire of CD8 T cells (2014)
  • Author(s): Keiji Tanaka
  • Organizer: EMBO Conference on Ubiquitin & ubiquitin-like proteins : At the crossroads from chromatin to protein.
  • Place of Presentation: Buenos Aires, Argentina(発表確定)
  • Year and Date: 20141019-24
  • Invited
• [Presentation] 神経変性疾患の謎を解く―タンパク分解系からのアプローチ (2014)
  • Author(s): 田中啓二
  • Organizer: 第37回日本神経科学会Neuroscience2014, 教育講演
  • Place of Presentation: パシフィコ横浜、横浜(発表確定)
  • Year and Date: 20140911-13
  • Invited
• [Presentation] 生化学の立場から (2014)
  • Author(s): 田中啓二
  • Organizer: 第61回日本栄養改善学会学術集会8/21メインシンポジウムI「栄養学―新たな挑戦 日本栄養改善学会60周年シンポジウム」
  • Place of Presentation: パシフィコ横浜、横浜(発表確定)
  • Year and Date: 20140820-22
  • Invited
• [Presentation] プロテアソーム : 動作原理から炎症免疫疾患制御への展望 (2014)
  • Author(s): 田中啓二
  • Organizer: 第35回日本炎症・再生医学会 : 教育講演
  • Place of Presentation: 万国津梁館、沖縄(発表確定)
  • Year and Date: 20140701-04
  • Invited
• [Presentation] 私のプロテアソーム研究を回想して… (2014)
  • Author(s): 田中啓二
  • Organizer: 第66回細胞生物学会大会 S11 七人の侍からのメッセージ～私の仕事を通じて若者達に伝えたきこと～
  • Place of Presentation: 奈良県新公会堂・東大寺総合文化センター、奈良(発表確定)
  • Year and Date: 20140611-13
  • Invited
• [Presentation] The PINK1-Parkin system controls mitochondrial homeostasis to suppress the cause of 'Parkinson's disease. (2014)
  • Author(s): Keiji Tanaka
  • Organizer: "KSU International Symposium : Cutting-edge of Life Sciences".
  • Place of Presentation: Kyoto, Japan(発表確定)
  • Year and Date: 20140530-31
  • Invited
• [Presentation] Basic mechanisms and physiopathological roles of eukaryotic proteasomes (2014)
  • Author(s): Keiji Tanaka
  • Organizer: A lecture in "Karolinska Research Lectures" series at Nobel Forum
  • Place of Presentation: Stockholm, Sweden(発表確定)
  • Year and Date: 2014-09-18
  • Invited
• [Presentation] タンパク質の新陳代謝～健康と病気のみなもと～ (2014)
  • Author(s): 田中啓二
  • Organizer: サイエンステクノフロンティアフォーラムサイテックサロン
  • Place of Presentation: 駒場ファカルティハウス(東京大学駒場キャンパス内、)東京(発表確定)
  • Year and Date: 2014-06-28
  • Invited
• [Presentation] Oxidative Stress, Mitophagy, and the PINK1-Parkin System (2014)
  • Author(s): Keiji Tanaka
  • Organizer: Symposium 17 on Oxidative Stress and Autophagy. 17th Biennial Meeting of Society for Free Radical Research International (SFRRI 2014)
  • Place of Presentation: Kyoto International Conference Center, Kyoto, Japan
  • Year and Date: 2014-03-26
  • Invited
• [Presentation] Selective Autophagy, Mitochondria, and Tumor Development (2013)
  • Author(s): Keiji Tanaka
  • Organizer: The 4th International Symposium on Dynamics of Mitochondria (DynaMito 2013)
  • Place of Presentation: Okinawa Zanpamisaki Royal Hotel, Okinawa, Japan
  • Year and Date: 20131012-1101
  • Invited
• [Presentation] In vivo interplay between the proteasome and autophagy in the mouse liver. (2013)
  • Author(s): Keiji Tanaka
  • Organizer: The 35th Naito Conference entitled The Ubiquitin-Proteasome. System : From Basic Mechanisms to Pathophysiological Roles.
  • Place of Presentation: Chateraise Gateaux Kingdom, Sapporo, Japan.
  • Year and Date: 20130709-12
  • Invited
• [Presentation] プロテアソームの発見から現在に至る研究の軌跡. (2013)
  • Author(s): 田中啓二
  • Organizer: 第29回 前立腺シンポジウム : 教育セミナー1
  • Place of Presentation: 東京コンファレンスセンター・品川、東京
  • Year and Date: 2013-12-14
  • Invited
• [Presentation] ミトコンドリアストレスとユビキチン・プロテアソームシステム (2013)
  • Author(s): 田中啓二
  • Organizer: 第8回臨床ストレス応答学会
  • Place of Presentation: 信州大学医学部、松本
  • Year and Date: 2013-11-16
  • Invited
• [Presentation] 私のプロテアソーム研究～成功と失敗の狭間で… (2013)
  • Author(s): 田中啓二
  • Organizer: 新学術研究「修飾シグナル病」第2回 若手ワークショップ 別講演
  • Place of Presentation: 伊香保温泉:雨情の湯 森秋旅館、群馬
  • Year and Date: 2013-10-22
  • Invited
• [Presentation] オートファジーKOマウスとその応用 (2013)
  • Author(s): 田中啓二, 小松雅明
  • Organizer: BioJapan 2013 World Business Forum
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2013-10-11
  • Invited
• [Presentation] The Keap1-Nrf2 activation during selective autophagy : its implication in tumor development. (2013)
  • Author(s): Keiji Tanaka and Masaaki Komatsu
  • Organizer: The 72nd Annual Meeting of Japanese Cancer Association Cancer research providing hope to fight against cancer Pacifico Yokohama Symposium 10 on Autophagy in Cancer.
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2013-10-03
  • Invited
• [Presentation] The Ubiquitin-Proteasome System. (2013)
  • Author(s): Keiji Tanaka
  • Organizer: HUPO 2013 Congress in Yokohama Proteomics of Protein Degradation : Session 11. Keynote Lecture
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2013-09-16
  • Invited
• [Presentation] Proteasome-over a quarter of a century from its discoveryプロテアソーム-その発見から四半世紀の軌跡 (2013)
  • Author(s): 田中啓二
  • Organizer: The 86th Annual Meeting of the Japanese Biochemical Society Globalizing Biochemistry's Future. Plenary Lecture
  • Place of Presentation: Yokohama (Japan)
  • Year and Date: 2013-09-13
  • Invited
• [Presentation] 二つの免疫型プロテアソームの発見-偶然から免疫学の檜舞台へ- (2013)
  • Author(s): 田中啓二
  • Organizer: 15th免疫サマースクール2013 in Hakata
  • Place of Presentation: 福岡
  • Year and Date: 2013-07-31
  • Invited
• [Presentation] プロテアソーム―発見から四半世紀の軌跡― (2013)
  • Author(s): Keiji Tanaka
  • Organizer: 大阪大学生命機能研究科研究交流会Frontier Bioscience Colloquium特別講演
  • Place of Presentation: 大阪
  • Year and Date: 2013-05-29
  • Invited
• [Presentation] Pathophysiological Roles of the Proteasome in Vertebrate. (2013)
  • Author(s): Keiji Tanaka
  • Organizer: The EUROMEDLAB Milano 2013 Congress. [OPENING LECTURE]
  • Place of Presentation: Milano Convention Center, Milano, Italy.
  • Year and Date: 2013-05-19
  • Invited
• [Remarks]
  • URL: http://www.igakuken.or.jp/pro-meta/
• [Patent(Industrial Property Rights)] ポリユビキチン化基質の同定方法 (2013)
  • Inventor(s): 吉田雪子, 佐伯 泰, 土屋 光, 村上有沙, 田中啓二
  • Industrial Property Rights Holder: (公財)東京都医学総合研究所
  • Industrial Property Rights Type: 特許, 特願
  • Industrial Property Number: 2013-237362
  • Filing Date: 2013-11-15
• [Patent(Industrial Property Rights)] A novel versatile method for determining ubiuuitin chain length reveals functional units of polyyubiguitin chains in cells (2013)
  • Inventor(s): 佐伯 泰, 土屋 光, 海保 愛, 田中啓二
  • Industrial Property Rights Holder: (公財)東京都医学総合研究所
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 61/901452
  • Filing Date: 2013-11-08
  • Overseas