Elucidation of the mechanism of physiological ER stress response required for the homeostasis of cell and whole body

• Project/Area Number: 17H01468
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: Kohno Kenji 奈良先端科学技術大学院大学, 研究推進機構, 特任教授 (50142005)
• Co-Investigator(Kenkyū-buntansha): 都留 秋雄 奈良先端科学技術大学院大学, 先端科学技術研究科, 助教 (80273861) ; 木俣 行雄 奈良先端科学技術大学院大学, バイオサイエンス研究科, 准教授 (60263448)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥43,030,000 (Direct Cost: ¥33,100,000、Indirect Cost: ¥9,930,000); Fiscal Year 2019: ¥13,130,000 (Direct Cost: ¥10,100,000、Indirect Cost: ¥3,030,000); Fiscal Year 2018: ¥11,570,000 (Direct Cost: ¥8,900,000、Indirect Cost: ¥2,670,000); Fiscal Year 2017: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
• Keywords: 小胞体 / ストレス応答 / インスリン / 糖尿病 / リボソーム / 翻訳休止 / 小胞体ストレス / 品質管理 / プロインスリン / 特殊スプライシング / 翻訳停止 / ストレス

Outline of Final Research Achievements

Firstly, the constitutive activation of IRE1-XBP1 pathway in the ER of mouse islet beta-cells is required for the normal insulin secretion and both the survival and the maintenance of mouse islet beta-cells using IRE1 knockout mice. Secondly, cryo-electron micrographic studies clarified the 3D structure of translational pausing peptide of XBP1u protein, indicating that the specific amino acid residues in XBP1u directly interacts with the ribosomal components in ribosome tunnel.

Academic Significance and Societal Importance of the Research Achievements

糖尿病の原因となる血糖値調節ホルモンのインスリン分泌が、膵島β細胞での小胞体ストレス応答経路IRE1-XBP1経路の恒常的活性化に依存していることを明らかにした。この結果は、正常なインスリン分泌の理解の一助になるとともに糖尿病治療にとっても役に立つ。また、XBP1蛋白質の翻訳過程での一時的休止は、上記の効率的な情報伝達に必要であるが、何故一時休止ということが起こるのか、その原因についても明らかにした。

Research Products

• [Int'l Joint Research] Ludwig-Maximilians-Universitat Munchen(ドイツ)
• [Int'l Joint Research] University of California, San Francisco(米国)
• [Int'l Joint Research] Ludwig-Maximilians-Universitat Munchen(ドイツ)
• [Int'l Joint Research] University of California, San Francisco/Harvard Medical School(米国)
• [Int'l Joint Research] University of Cambridge(United Kingdom)
• [Int'l Joint Research] Pohang Univ. of Science & Technology(韓国)
• [Journal Article] Structural and mutational analysis of the ribosome-arresting human XBP1u (2019)
  • Author(s): Shanmuganathan, V.
  • Journal Title: eLife Volume: 8
  • DOI: 10.7554/elife.46267
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] IRE1-XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells (2018)
  • Author(s): Yuichi Tsuchiya, Michiko Saito, Hiroshi Kadokura, Jun-ichi Miyazaki, Fumi Tashiro, Yusuke Imagawa, Takao Iwawaki, Kenji Kohno
  • Journal Title: The Journal of Cell Biology Volume: 217 Issue: 4 Pages: 1287-1301
  • DOI: 10.1083/jcb.201707143
  • Peer Reviewed / Open Access
• [Journal Article] 4-Phenylbutyrate suppresses the unfolded protein response without restoring protein folding in Saccharomyces cerevisiae (2018)
  • Author(s): Mai Chi Thanh、Le Quynh Giang、Ishiwata-Kimata Yuki、Takagi Hiroshi、Kohno Kenji、Kimata Yukio
  • Journal Title: FEMS Yeast Research Volume: 18 Issue: 2 Pages: 1-8
  • DOI: 10.1093/femsyr/foy016
  • NAID: 120006537413
  • Peer Reviewed
• [Journal Article] Identification of the physiological substrates of PDIp, a pancreas-specific protein-disulfide isomerase family member (2018)
  • Author(s): Fujimoto Takushi、Nakamura Orie、Saito Michiko、Tsuru Akio、Matsumoto Masaki、Kohno Kenji、Inaba Kenji、Kadokura Hiroshi
  • Journal Title: Journal of Biological Chemistry Volume: 293 Issue: 48 Pages: 18421-18433
  • DOI: 10.1074/jbc.ra118.003694
  • Peer Reviewed / Open Access
• [Journal Article] 小胞体膜への蛋白質の標的化―蛋白質の個性に応じた輸送経路 (2018)
  • Author(s): 大古殿美加、河野憲二
  • Journal Title: 医学のあゆみ Volume: 267 Pages: 971-975
• [Journal Article] 小胞体ストレスセンサーIRE1αによるプロインスリンの酸化的折り畳みの制御機構 (2018)
  • Author(s): 土屋雄一、河野憲二
  • Journal Title: 化学と生物 Volume: 57 Pages: 161-166
  • Peer Reviewed
• [Journal Article] Identification and functional study of ER stress sensor IRE1 in Chlamydomonas reinhardtii (2018)
  • Author(s): Yasuyo Yamaoka
  • Journal Title: The Plant Journal Volume: 94巻 Issue: 1 Pages: 91-104
  • DOI: 10.1111/tpj.13844
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 4μ8C Inhibits Insulin Secretion Independent of IRE1α RNase Activity (2017)
  • Author(s): Sato, H., Shiba, Y., Tsuchiya, Y., Saito, M., Kohno, K.
  • Journal Title: Cell Structure and Function Volume: 42 Issue: 1 Pages: 61-70
  • DOI: 10.1247/csf.17002
  • NAID: 130005635485
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Ectopic expression of the transcription factor MafB in basal keratinocytes induces hyper-proliferation and perturbs epidermal homeostasis. (2017)
  • Author(s): Miyai M, Tsunekage Y, Saito M, Kohno K, Takahashi K, Kataoka K.
  • Journal Title: Experimental Dermatology Volume: 26 Issue: 11 Pages: 1039-1045
  • DOI: 10.1111/exd.13364
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Defective ATG16L1-mediated removal of IRE1α drives Crohn’s disease-like ileitis. (2017)
  • Author(s): Tschurtschenthaler, M., Adolph, T.E., Ashcroft, J.W., Niederreiter, L., Bharti, R., Saveljeva, S., Bhattacharyya, J., Flak, M.B., Shih, D.Q., Fuhler, G.M., Parkes, M., Kohno, K., Iwawaki, T., van der Woude, C.J., Harding, H., Smith, Peppelenbosch, M.P., Targan, S.R., Ron, D., Rosenstiel, P., Blumberg, R.S., Kaser, A.
  • Journal Title: J. Exp. Med. Volume: 214 Issue: 2 Pages: 401-422
  • DOI: 10.1084/jem.20160791
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 重複機能ドメインの解析から得られたXBP1遺伝子の起源と進化 (2019)
  • Author(s): 小池雅昭
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] XBP1u翻訳休止におけるリボソームタンパク質uL4の関与 (2019)
  • Author(s): 大古殿美加 曽川愛守榮 木俣行雄 河野憲二
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] ヒト細胞小胞体機能の低下を鋭敏に検出するレポーターの開発と応用 (2019)
  • Author(s): 八巻聡
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] XBP1u translational pausing requires the interaction between own nascent chain and uL4 (2019)
  • Author(s): 大古殿美加
  • Organizer: 第71回日本細胞生物学会
• [Presentation] Conserved overlapping coding frame regulates two types of XBP1 functions. (2018)
  • Author(s): 小池雅俊
  • Organizer: 新学術若手ワークショップ
• [Presentation] みなしごサブユニットに対する品質管理因子の発見 (2018)
  • Author(s): 柳谷耕太
  • Organizer: 新学術若手ワークショップ
  • Invited
• [Presentation] IRE1α-XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells. (2018)
  • Author(s): Tsuchiya Yuichi, Saito Michiko, Kadokura Hiroshi, Miyazaki Jun-ichi, Tashiro Fumi, Imagawa Yusuke, Iwawaki Takao, Kohno Kenji
  • Organizer: 第70回日本細胞生物学会
• [Presentation] Conserved overlapping coding frame regulates two types of XBP1 functions. (2018)
  • Author(s): Masaaki Koike
  • Organizer: 第70回日本細胞生物学会
• [Presentation] The role and the mechanism of XBP1u-translational pausing in the ER stress signaling. (2018)
  • Author(s): Kenji Kohno
  • Organizer: International Symposium; Proteins: From the cradle to the grave
  • Int'l Joint Research / Invited
• [Presentation] Requirement of ER stress response for nematode expulsion through mature mucin production by IRE1β specific pathway. (2018)
  • Author(s): Akio Tsuru
  • Organizer: International Symposium; Proteins: From the cradle to the grave
  • Int'l Joint Research
• [Presentation] XBP1 encodes SRP recognition motif and transactivation domain in a conserved dual-coding frame. (2018)
  • Author(s): Masaaki Koike
  • Organizer: International Symposium; Proteins: From the cradle to the grave
  • Int'l Joint Research
• [Presentation] IRE1α-XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells. (2018)
  • Author(s): Yuichi Tsuchiya
  • Organizer: International Symposium; Proteins: From the cradle to the grave
  • Int'l Joint Research
• [Presentation] XBP1u tranlational pausing requires the interaction between own nascent chain and ribosomal proteins. (2018)
  • Author(s): Miku Ohfurudono
  • Organizer: International Symposium; Proteins: From the cradle to the grave
  • Int'l Joint Research
• [Presentation] The role and mechanism of XBP1u-translational pausing in the ER stress signaling. (2018)
  • Author(s): Kenji Kohno
  • Organizer: EMBO workshop: Endoplasmic Reticulum function and disease
  • Int'l Joint Research
• [Presentation] XBP1u翻訳休止機構と小胞体ストレス下での役割 (2018)
  • Author(s): Kenji Kohno
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] Roles of listerin in protein translocation into the endoplasmic reticulum. (2018)
  • Author(s): Ashway Sogawa
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] みなしご蛋白質に対する品質管理因子UBE2Oの発見 (2018)
  • Author(s): Kota Yanagitani
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] Molecular mechanism of XBP1u translational pausing (2017)
  • Author(s): Miku Ohfurudono, Ash-way Sogawa, Yukio Kimata, Kenji Kohno
  • Organizer: EMBO Conference: Protein quality control
  • Int'l Joint Research
• [Presentation] Translational pausing in the ER stress signaling (2017)
  • Author(s): Kenji Kohno
  • Organizer: International Symposium on Protein Quality Control
  • Int'l Joint Research / Invited
• [Presentation] Translational pausing in the ER stress signaling (2017)
  • Author(s): Kenji Kohno
  • Organizer: FASEB Science Research Conferences
  • Int'l Joint Research / Invited
• [Presentation] The role of translational pausing in the delivery of XBP1u mRNA to the ER via the SRP pathway (2017)
  • Author(s): Masaaki Koike, Satoshi Kanda, Kota Yanagitani, Kenji Kohno
  • Organizer: 第17回日本蛋白質科学会ワークショッップ
  • Invited
• [Presentation] Unfolded protein response-オルガネラ異常検知メカニズム- (2017)
  • Author(s): 木俣行雄
  • Organizer: 日本植物学会第81回大会シンポジウム
  • Invited
• [Presentation] XBP1sタンパク質における転写活性化ドメインの同定 (2017)
  • Author(s): 小池雅昭, 野坂祐, 河野憲二
  • Organizer: 第12回小胞体ストレス研究会
• [Presentation] ムチン産生に関る小胞体ストレスセンサーIRE1βはIRE1αとは異なるリン酸化制御を受ける (2017)
  • Author(s): 都留秋雄, 坪山恭之, 大澤祐介, 河野憲二
  • Organizer: 第12回小胞体ストレス研究会
• [Presentation] 膵島β細胞でのIRE1α-XBP1経路の生理的活性化の意義 (2017)
  • Author(s): 土屋雄一, 斉藤美知子, 門倉広, 宮崎純一, 田代文, 今川佑介, 岩脇隆夫, 河野憲二
  • Organizer: 第12回小胞体ストレス研究会
• [Presentation] Activation of yeast Ire1 upon diauxic shift (2017)
  • Author(s): Tran Minh Duc, 木俣行雄
  • Organizer: 第12回小胞体ストレス研究会
• [Presentation] ペプチド鎖伸長因子2に形成されるジフタミドの生理的意義の解明 (2017)
  • Author(s): 小池雅昭, 額田知実, 山田麗奈, 赤沼啓志, 斉藤美知子, 伊川正人, 河野憲二
  • Organizer: 第69回日本細胞生物学会
• [Presentation] 小胞体ストレスセンサーIRE1βの活性化においてリン酸化が果たす複数の役割 (2017)
  • Author(s): 都留秋雄, 坪山恭之, 大澤祐介, 河野憲二
  • Organizer: 生命科学系学会合同年次大会（ConBio2017)
• [Presentation] 哺乳動物細胞におけるXBP1u の機能性翻訳休止機構の解明 (2017)
  • Author(s): 大古殿美加, 曽川愛守榮, 木俣行雄, 河野憲二
  • Organizer: 生命科学系学会合同年次大会（ConBio2017)
• [Book] Stress response mechanisms in fungi-theoretical and practical aspects (2019)
  • Author(s): Kimata, Y., Nguyen, T.M.P., Kohno, K.
  • Publisher: Springer
• [Remarks] 河野特任研究プロジェクト
  • URL: http://www.naist.jp/iri/kouno/
• [Remarks] 河野特任研究プロジェクト
  • URL: https://www.facebook.com/naist.kohnolab/