ミトコンドリアを介したストレス応答におけるタンパク質切断とリン酸化の役割

• Project Area: Regulation of signal transduction by post-translational modifications and its pathogenic dysregulation
• Project/Area Number: 23117510
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagasaki University (2012); The University of Tokyo (2011)
• Principal Investigator: 武田 弘資 長崎大学, 医歯(薬)学総合研究科, 教授 (10313230)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000); Fiscal Year 2012: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2011: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
• Keywords: ミトコンドリア / ストレス応答 / ホスファターゼ / プロテアーゼ

Outline of Annual Research Achievements

PGAM5のミトコンドリア内での局在について免疫電顕を用いて解析したところ、前年度に行った生化学的な解析結果と一致して、おもに内膜に局在することが確認された。また、脱共役剤CCCP処理によって膜電位を失ったミトコンドリアにおいて、切断されたPGAM5がミトコンドリア内に保持されていることも分かった。今後、細胞死誘導刺激時などに、切断されたPGAM5の局在が変化するかを検討する予定である。
一方、PGAM5 KOマウスより単離した骨髄由来マクロファージの応答を野生型マウス由来のマクロファージと比較・検討する過程で、Nigericinと呼ばれる細菌性毒素による炎症性サイトカインIL-1#223;の産生が、KOマウス由来のマクロファージで低下していることを見出した。IL-1#223;の産生・分泌は、インフラマソームと呼ばれる細胞内タンパク質複合体によって担われており、NigericinはそのうちNLRP3分子を構成因子とするインフラマソーム（NLRP3インフラマソーム）の活性化因子として知られている。Nigericin以外のいくつかのNLRP3インフラマソーム活性化刺激に対しては、PGAM5 KOマウス由来のマクロファージでのIL-1#223;の産生低下は認められなかったことから、PGAM5はNigericinによるNLRP3インフラマソームの活性化に特異的に働いていることが示唆される。現在、NLRP3インフラマソームの活性化におけるPGAM5の役割について、ホスファターゼ活性や膜内切断との関連を中心に検討を進めている。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Atypical protein phosphatases: emerging players in cellular signaling. (2013)
  • Author(s): Sadatomi, D. et al.
  • Journal Title: Int. J. Mol. Sci. Volume: 14 Issue: 3 Pages: 4596-4612
  • DOI: 10.3390/ijms14034596
  • Peer Reviewed
• [Journal Article] A novel monoclonal antibody reveals a conformational alteration shared by amyotrophic lateral sclerosis-linked SOD1 mutants. (2012)
  • Author(s): Fujisawa, T., Homma, K., et al.
  • Journal Title: Ann Neurol Volume: 72 Issue: 5 Pages: 739-749
  • DOI: 10.1002/ana.23668
  • Peer Reviewed
• [Journal Article] Rhomboid protease PARL mediates the mitochondrial membrane potential loss-induced cleavage of PGAM5 (2012)
  • Author(s): Sekine, S., et al.
  • Journal Title: J. Biol. Chem. Volume: 287 Issue: 41 Pages: 34635-34645
  • DOI: 10.1074/jbc.m112.357509
  • Peer Reviewed
• [Journal Article] Apoptosis signal-regulating kinase-1 inhibitor as a potent therapeutic drug for the treatment of gastric cancer. (2012)
  • Author(s): Hayakawa, Y., Hirata, Y., Sakitani, K., Nakagawa, H., Nakata, W., Kinoshita, H., Takahashi, R., Takeda, K., Ichijo, H., Maeda1, S. and Koike, K.
  • Journal Title: Cancer Sci. Volume: 103 Issue: 12 Pages: 2181-2185
  • DOI: 10.1111/cas.12024
  • Peer Reviewed
• [Journal Article] The kelch repeat protein KLHDC10 regulates oxidative stress-induced ASK1 activation by suppressing PP5 (2012)
  • Author(s): Yusuke Sekine, Ryo Hatanaka, Takeshi Watanabe. Naoki Sono, Shun-ichiro Iemura, Tohru Natsume, Erina Kuranaga, Masayuki Miura, Kohsuke Takeda, and Hidenori Ichijo
  • Journal Title: Molecular Cell Volume: 48 Issue: 5 Pages: 1-13
  • DOI: 10.1016/j.molcel.2012.09.018
  • NAID: 120007150217
  • Peer Reviewed
• [Journal Article] ASK3 responds to osmotic stress and regulates blood pressure by suppressing WNK1-SPAK/OSR1 signaling in the kidney. (2012)
  • Author(s): Naguro I., et al.
  • Journal Title: Nature Communications Volume: 3 Issue: 1 Pages: 1285-1285
  • DOI: 10.1038/ncomms2283
  • Peer Reviewed
• [Journal Article] The Phosphorylation-Dependent Regulation of Mitochondrial Proteins in Stress Responses (2012)
  • Author(s): Kanamaru, Y., Sekine S., Ichijo, H. and Takeda, K.
  • Journal Title: J. Signal Transduc. Volume: 2012 Pages: 12-12
  • DOI: 10.1155/2012/931215
  • Peer Reviewed
• [Journal Article] Therapeutic targets in the ASK1-dependent stress signaling pathways (2012)
  • Author(s): Hayakawa, R., Hayakawa, T., Takeda, K. and Ichijo, H.
  • Journal Title: Proceedings of the Japan Academy, Series B Volume: 88 Issue: 8 Pages: 434-453
  • DOI: 10.2183/pjab.88.434
  • NAID: 130001924759
  • ISSN: 0386-2208, 1349-2896
  • Peer Reviewed
• [Journal Article] Prevention of apoptosis by mitochondrial phosphatase PGAM5 in the mushroom body is crucial for heat shock resistance in Drosophila melanogaster (2012)
  • Author(s): Ishida, Y., et al.
  • Journal Title: PloS ONE Volume: 7 Issue: 2 Pages: 3006-3016
  • DOI: 10.1371/journal.pone.0030265
  • Peer Reviewed
• [Journal Article] p38 MAP kinase regulates the expression of genes in the dopamine synthesis pathway through phosphorylation of NR4A nuclear receptors (2011)
  • Author(s): Sekine,Y., Takagahara,S., Hatanaka, R., Watanabe, T., Oguchi, H., Noguchi, T., Naguro, I., Kobayashi, K., Tsunoda, M., Funatsu, T., Nomura, H., Toyoda, T., Matsuki, N., Kuranaga, E., Miura, M., Takeda, K., and Ichijo, H.
  • Journal Title: J. Cell Sci. Volume: 124 Issue: 17 Pages: 3006-3016
  • DOI: 10.1242/jcs.085902
  • Peer Reviewed
• [Journal Article] Apoptosis signal-regulating kinase 1 inhibits hepatocarcinogenesis by controlling the tumor-suppressing function of stress-activated MAPK (2011)
  • Author(s): Nakagawa, H.
  • Journal Title: Hepatology Volume: 54 Issue: 1 Pages: 185-195
  • DOI: 10.1002/hep.24357
  • Peer Reviewed
• [Journal Article] Identification of novel ASK1 inhibitors using virtual screening (2011)
  • Author(s): Okamoto, M., et al
  • Journal Title: Bioorg.Med.Chem. Volume: 19 Issue: 1 Pages: 486-489
  • DOI: 10.1016/j.bmc.2010.11.004
  • Peer Reviewed
• [Journal Article] Regulation of anoxic death in Caenorhabditis elegans by mammalian apoptosis signal-regulating kinase (ASK) family proteins (2011)
  • Author(s): Hayakawa, T., Kato, K., Hayakawa, R., Hisamoto, N., Matsumoto, K., Takeda, K. and Ichijo, H.
  • Journal Title: Genetics Volume: 187 Issue: 3 Pages: 785-792
  • DOI: 10.1534/genetics.110.124883
  • Peer Reviewed
• [Journal Article] Apoptosis signal-regulating kinase 1 and cyclin D1 compose a positive feedback loop contributing to tumor growth in gastric cancer (2011)
  • Author(s): Hayakawa, Y., et al
  • Journal Title: Proc.Natl.Acad.Sci.USA. Volume: 108 Issue: 2 Pages: 780-785
  • DOI: 10.1073/pnas.1011418108
  • Peer Reviewed
• [Journal Article] Apoptosis signaling kinases: from stress response to health outcomes (2011)
  • Author(s): Takeda, K., Naguro, I., Nishitoh, H., Matsuzawa, A. and Ichijo, H.
  • Journal Title: Antioxid. Redox Signal. Volume: 15 Issue: 3 Pages: 719-761
  • DOI: 10.1089/ars.2010.3392
  • Peer Reviewed
• [Presentation] Role of mitochondrial phosphatase PGAM5 in inflammation (2013)
  • Author(s): Takeda, K.
  • Organizer: JST-CREST International Symposium, Frontiers in Immunology and Inflammation: From Molecules to Disease
  • Place of Presentation: 東京
  • Year and Date: 2013-02-12
• [Presentation] Regulation of cellular stress response by mitochondrial protein phosphatase PGAM5 (2013)
  • Author(s): Takeda, K.
  • Organizer: 1st International Symposium on Protein Modifications in Pathogenic Dysregulation of Signaling
  • Place of Presentation: 東京
• [Presentation] Role of mitochondrial protein phosphatase PGAM5 in inflammation (2013)
  • Author(s): Sadatomi, D., Takeda, K.
  • Organizer: 10th International Conference on Protein Phosphatase
  • Place of Presentation: 東京
• [Presentation] ミトコンドリアのストレス受容・応答機構と炎症制御 (2013)
  • Author(s): 武田 弘資
  • Organizer: 創薬シンポジウム：アカデミア創薬と探索医療
  • Place of Presentation: 長崎
• [Presentation] ミトコンドリア機能低下の感知システムとしてのプロテインホスファターゼ PGAM5の膜内切断 (2013)
  • Author(s): 武田 弘資
  • Organizer: 日本薬学会第133年会
  • Place of Presentation: 横浜
• [Presentation] Role of mitochondrial phosphatase PGAM5 in inflammation (2012)
  • Author(s): Takeda, K.
  • Organizer: The 12th Biennial International Endotoxin Innate Immunity Society Meeting (IEIIS2012)
  • Place of Presentation: 東京
  • Invited
• [Presentation] KLHDC10によるPP5の活性制御と酸化ストレス応答 (2012)
  • Author(s): 武田弘資, 他
  • Organizer: 第5回日本プロテインホスファターゼ研究会学術集会
  • Place of Presentation: 大阪
• [Presentation] Analysis of ASK1 in contact hypersensitivity using a chemical genetic approach (2012)
  • Author(s): Takeda, K., et al
  • Organizer: The 4th Global COE Retreat in Oiso
  • Place of Presentation: 大磯
• [Presentation] Regulation of cellular stress response by an atypical protein ghosphatase PGAM5 (2011)
  • Author(s): Takeda, K
  • Organizer: The First Japan-Taiwan Bilateral Conference on Protein Phosphatases
  • Place of Presentation: 岡崎
  • Year and Date: 2011-12-02
• [Presentation] アティピカル・プロテインホスファターゼPGAM5によるストレス応答制御 (2011)
  • Author(s): 武田弘資
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
• [Book] Roles for the stress-responsive kinases ASK1 and ASK2 in tumorigenesis. Chembiomolecular Science: At the Frontier of Chemistry and Biology, eds Shibasaki, M., Iino, M. and Osada, H. (2012)
  • Author(s): Kamiyama, M. et al.
  • Total Pages: 325
  • Publisher: Springer Japan
• [Book] がんと炎症．「がん増殖と悪性化の分子機構」 (2012)
  • Author(s): 武田 弘資
  • Total Pages: 280
  • Publisher: 化学同人
• [Remarks]
  • URL: http://www.ph.nagasaki-u.ac.jp/lab/cell/index-j.html