Structural analysis of proteasome and PI31, a regulator of the activity and the molecular assembly of proteasome.

• Project/Area Number: 25870631
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry; Functional biochemistry
• Research Institution: University of Hyogo
• Principal Investigator: TAKAGI Kenji 兵庫県立大学, 生命理学研究科, 助教 (90647322)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2013: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: X線結晶構造解析 / 細胞内タンパク質分解 / プロテアソーム / 分子集合 / タンパク質分解酵素

Outline of Final Research Achievements

PI31 plays regulatory roles of proteasome subunit assembly and substrate degradation activity. To elucidate the details of proteasomal regulation by PI31, we carried out studies on expression and purification of PI31, and structural analysis of 20S CP α7ΔN mutant, which complement PI31 defection. Based on the revealed structure we looked at working mechanism of PI31. In the 20S CP α7ΔN, N-terminal peptide differs from it in wild type structure. This difference is thought to change the α1 orientation and affect its proteasome function.

Research Products

• [Journal Article] N-terminal α7 deletion of the proteasome 20S core particle substitutes for yeast PI31 function. (2015)
  • Author(s): Yashiroda H, Toda Y, Otsu S, Takagi K, Mizushima T, Murata S.
  • Journal Title: Mol Cell Biol Volume: 35(1) Issue: 1 Pages: 141-52
  • DOI: 10.1128/mcb.00582-14
  • Peer Reviewed
• [Journal Article] Structural determinants in GABARAP required for the selective binding and recruitment of ALFY to LC3B-positive structures. (2014)
  • Author(s): Lystad, AH., Ichimura, Y., Takagi, K., Yang, Y., Pankiv, S., Kanegae, Y., Kageyama, S., Suzuki, M., Saito, I., Mizushima,T., *Komatsu, M., *Simonsen, A.
  • Journal Title: EMBO Rep. Volume: 15 Issue: 5 Pages: 557-565
  • DOI: 10.1002/embr.201338003
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Pba3-Pba4 heterodimer acts as a molecular matchmaker in proteasome α-ring formation (2014)
  • Author(s): Takagi, K., Saeki, Y., Yashiroda, H., Yagi, H., Kaiho, A., Murata, S., Yamane, T., Tanaka, K., Mizushima, T. and Kato, K.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 450 Issue: 2 Pages: 1110-1114
  • DOI: 10.1016/j.bbrc.2014.06.119
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Bacterial Effectors and Their Functions in the Ubiquitin-Proteasome System: Insight from the Modes of Substrate Recognition (2014)
  • Author(s): Kim M et. al
  • Journal Title: Cells Volume: 3 Issue: 3 Pages: 848-864
  • DOI: 10.3390/cells3030848
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Kinetic, Thermodynamic and Structural Characterizations of Association between Nrf2-DLGex Degron and Keap1. (2014)
  • Author(s): Fukutomi, T., Takagi, K., Mizushima, T., Ohuchi, N., and Yamamoto, M.
  • Journal Title: Moll Cell Biol. Volume: 34 Issue: 5 Pages: 832-846
  • DOI: 10.1128/mcb.01191-13
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structural basis for the recognition of Ubcl3 by the Shigell a flexneri effector OspI (2013)
  • Author(s): Nishide A, et. al
  • Journal Title: J Mol Bio. Volume: 425 Issue: 15 Pages: 2623-2631
  • DOI: 10.1016/j.jmb.2013.02.037
  • 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 Issue: 5 Pages: 618-631
  • DOI: 10.1016/j.molcel.2013.08.003
  • Peer Reviewed / Open Access
• [Presentation] 赤痢菌エフェクターIpaH9.8基質認識ドメインのX線結晶構造解析 (2014)
  • Author(s): 高木賢治、Kim Minsoo、笹川千尋、水島恒裕
  • Organizer: 新学術領域研究「ユビキチンネオバイオロジー」平成26年度領域班会議
  • Place of Presentation: ホテルたつき（愛知県蒲郡市）
  • Year and Date: 2014-12-03 – 2014-12-05
• [Presentation] Structure and substrate recognition mechanism of IpaH9.8 LRR (2014)
  • Author(s): Kenji Takagi, Akira Nishide, Tsunehiro Mizushima
  • Organizer: Symposium for young ubiquitin researchers ina Japan "New Era in the Ubiquitin Research"
  • Place of Presentation: 国際高等研究所（京都府木津川市）
  • Year and Date: 2014-11-11 – 2014-11-12
• [Presentation] 赤痢菌エフェクターIpaH9.8基質認識ドメインのX線結晶構造解析 (2014)
  • Author(s): 高木賢治、水島恒裕
  • Organizer: 日本結晶学会平成26年度年会
  • Place of Presentation: 東京大学（東京都文京区）
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] Structural basis for specific recognition of Rpt1 by proteasome-dedicated chaperone Hsm3 (2013)
  • Author(s): Kenji Takagi, Sangwoo Kim, Yasushi Saeki, Keiji Tanaka, Koichi Kato, Tsunehiro Mizushima
  • Organizer: 第35回内藤コンファレンス "The Ubiquitin-Proteasome System: From BasicMechanisms to Pathophysiological Roles"
  • Place of Presentation: シャトレーゼ ガトーキングダム サッポロ（北海道札幌市）
• [Presentation] 脱アミド化Ubc13の構造解析による赤痢菌感染機構の考察 (2013)
  • Author(s): 高木 賢治, 西出 旭, Kim Minsoo, 笹川 千尋, 水島 恒裕
  • Organizer: 新学術領域研究「ユビキチン制御」平成25年度第2回領域会議
  • Place of Presentation: 熱海ニューフジヤホテル（静岡県熱海市）
• [Book] プロテアソーム複合体形成シャペロンの構造と作用機構 (2013)
  • Author(s): 高木賢治, 水島恒裕
  • Publisher: 生化学
• [Book] Autophagy : Mechanisms of Regulation of p62 in Autophagy and Implications for Health and Diseases (2013)
  • Author(s): Kenji Takagi, Tsunehiro Mizushima
  • Publisher: Academic press