Identification and structural-functional analysis of shuttle-type proteasome activator exhibiting molecular chaperone activity

2013 Fiscal Year Final Research Report

• Project/Area Number: 24657113
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Biophysics
• Research Institution: Okazaki Research Facilities, National Institutes of Natural Sciences
• Principal Investigator: KATO Koichi 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 教授 (20211849)
• Co-Investigator(Renkei-kenkyūsha): YAMAGUCHI Takumi 分子科学研究所, 生命・錯体分子科学研究領域, 助教 (60522430) ; YAGI Maho 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 特任助教 (40608999) ; SATOH Tadashi 名古屋市立大学, 薬学研究科(研究院), 准教授 (80532100)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: プロテアソーム / 4次構造 / 活性化因子 / シャペロン / 古細菌

Research Abstract

Assembly of the eukaryotic 20S proteasome is not spontaneous self-organization but an ordered process involving several assembly chaperones, whereas that of the archaeal 20S proteasome involves spontaneous self-assembly. Recent genomic analysis identified archaeal homologs of the assembly chaperones, PbaA and PbaB. However, it remains unclear how such assembly chaperone-like proteins play an indispensable role in assembly of the proteasome subunit in archaea. This study revealed that PbaB actually functions as a proteasome activator. Furthermore, our integrative biochemical and biophysical approach including X-ray crystallography, electron microscopy, NMR spectroscopy, and small-angle neutron scattering provided mechanistic clues to the molecular action of the active complex formed between the PbaB homotetramer and 20S proteasome.

Research Products

• [Journal Article] Structural basis for proteasome formation controlled by an assembly chaperone Nas2 (2014)
  • Author(s): T. Satoh, Y. Saeki, T. Hiromoto, Y.-H. Wang,Y. Uekusa, H. Yagi, H. Yoshihara, M.Yagi-Utsumi, T. Mizushima, K. Tanaka, and K.Kato
  • Journal Title: Structure Volume: (in press)
  • DOI: 10.1016/j.str.2014.02.014
  • Peer Reviewed
• [Journal Article] Total synthesis and characterization of thielocin B1 as a protein–protein interaction inhibitor of PAC3 homodimer (2014)
  • Author(s): T. Doi, M. Yoshida, K. Ohsawa, K. Shin-ya,M. Takagi, Y. Uekusa, T. Yamaguchi, K. Kato,T. Hirokawa, and T. Natsume
  • Journal Title: Chem. Sci. Volume: 5 Pages: 1860-1868
  • DOI: 10.1039/C3SC53237B
  • Peer Reviewed
• [Journal Article] Conformational characterization of a protein complex involving intrinsically disordered protein by small-angle neutron scattering using the inverse contrast matching method : a case study of interaction betweenα-synuclein and PbaB tetramer as a model chaperone (2014)
  • Author(s): M. Sugiyama, H. Yagi, T. Yamaguchi, K.Kumoi, M. Hirai, Y. Oba, N. Sato, L. Porcar,A. Martele, and K. Kato
  • Journal Title: J. Appl. Cryst. Volume: 47 Pages: 430-435
  • DOI: 10.1107/S1600576713033475
  • Peer Reviewed
• [Journal Article] An archaeal homolog of proteasome assembly factor functions as a proteasome activator (2013)
  • Author(s): K. Kumoi, T. Satoh, K. Murata, T. Hiromoto, T. Mizushima, Y. Kamiya, M. Noda, S. Uchiyama, H. Yagi, and K. Kato
  • Journal Title: PLoS ONE Volume: 8 Pages: e60294
  • DOI: 10.1371/journal.pone.0060294
  • Peer Reviewed
• [Journal Article] Spatial arrangement and functional role of α subunits of proteasome activator PA28 in hetero-oligomeric form (2013)
  • Author(s): M. Sugiyama, H. Sahashi, E. Kurimoto, S.Takata, H. Yagi, K. Kanai, E. Sakata, Y.Minami, K. Tanaka, and K. Kato
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 432 Pages: 141-145
  • DOI: 10.1016/j.bbrc.2013.01.071
  • Peer Reviewed
• [Journal Article] New crystal structure of the proteasome-dedicated chaperone Rpn14 at 1.6Å resolution (2012)
  • Author(s): S. Kim, A. Nishide, Y. Saeki, K. Takagi, K.Tanaka, K. Kato, and T. Mizushima
  • Journal Title: Acta Cryst. Volume: F68 Pages: 517-521
  • DOI: 10.1107/S1744309112011359
  • Peer Reviewed
• [Journal Article] Structural basis for specific recognition of Rpt1, an ATPase subunit of the 26 S proteasome, by the proteasome-dedicated chaperone Hsm3 (2012)
  • Author(s): K. Takagi, S. Kim, H. Yukii, M. Ueno, R. Morishita, Y. Endo, K. Kato, K. Tanaka, Y. Saeki, and T. Mizushima
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 12172-12182
  • DOI: 10.1074/jbc.M112.345876
  • Peer Reviewed
• [Presentation] 生命分子の揺らぎと秩序形成独 (2014)
  • Author(s): 加藤晃一
  • Organizer: 日本学術振興会分子系の複合電子機能第181 委員会
  • Place of Presentation: 国際高等研究所(京都)
  • Year and Date: 2014-02-25
• [Presentation] Mechanistic Insights into Dynamic Orchestration of Proteasomes (2014)
  • Author(s): K. Kato
  • Organizer: Pure and Applied Chemistry International Conference (PACCON) 2014
  • Place of Presentation: Khon Kaen(Thailand)
  • Year and Date: 2014-01-08
• [Presentation] An archaeal homolog of proteasome assembly chaperone forms a homotetramer and functions as proteasome activator (2013)
  • Author(s): K. Kumoi, T. Satoh, K. Murata, T.Hiromoto, T. Mizushima, Y. Kamiya, M.Noda, S. Uchiyama, M. Sugiyama, H. Yagi,and K. Kato
  • Organizer: 6^<th> Japan-Korea Seminars on Biomolecular Science : Experiments and Simulation
  • Place of Presentation: 岡崎コンファレンスセンター(愛知)
  • Year and Date: 2013-11-27
• [Presentation] Structural basis for proteasome formation mediated by assembly chaperone Nas2 (2013)
  • Author(s): T. Satoh, H. Yagi, Y-H. Wang, Y. Uekusa,T. Hiromoto, M. Yagi-Utsumi, T.Mizushima, and K. Kato
  • Organizer: 6 ^<th> Japan-Korea Seminars on Biomolecular Science : Experiments and Simulation
  • Place of Presentation: 岡崎コンファレンスセンター(愛知)
  • Year and Date: 2013-11-27
• [Presentation] Interaction mode of a proteasome assembly chaperone Nas2 with the proteasome Rpt5 subunit (2013)
  • Author(s): Y-H. Wang, T. Satoh, T. Hiromoto, Y.Uekusa, H. Yagi, M. Yagi-Utsumi, T.Mizushima, Y. Saeki, K. Tanaka, and K.Kato
  • Organizer: 山田研究会・統合バイオサイエンスシンポジウム
  • Place of Presentation: 伊良湖
  • Year and Date: 2013-11-20
• [Presentation] Mechanistic insights into the proteasome orchestration by a proteasome assembly chaperone Nas2 (2013)
  • Author(s): T. Satoh, Y. Uekusa, T. Hiromoto, H. Yagi,M. Yagi-Utsumi, T. Mizushima, Y. Saeki,K. Tanaka, and K. Kato
  • Organizer: 7th International Conference on StructuralGenomics (ICSG2013-SLS)
  • Place of Presentation: 札幌
  • Year and Date: 2013-08-01
• [Presentation] Structural insights into proteasome orchestration mechanism through a proteasome assembly factor Nas2 (2013)
  • Author(s): T. Satoh, Y. Uekusa, T. Hiromoto, H. Yagi,M. Yagi-Utsumi, T. Mizushima, Y. Saeki,K. Tanaka, and K. Kato
  • Organizer: 8 ^<th>Asian Biophysics Association Symposium (ABA Jeju 2013)
  • Place of Presentation: Jeju, Korea
  • Year and Date: 2013-05-27
• [Presentation] プロテアソームの分子集合における集合因子Nas2 の機能発現機構 (2013)
  • Author(s): 佐藤匡史、植草義徳、廣本武史、矢木宏和、矢木真穂、水島恒裕、佐伯泰、田中啓二、加藤晃一
  • Organizer: 第77回日本生化学会中部支部例会・シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2013-05-25
• [Presentation] 生命分子のダイナミクスと自己組織化国際高等研究所研究プロジェクト (2013)
  • Author(s): 加藤晃一
  • Organizer: 「分子基盤に基づく生体機能への揺らぎとダイナミックネットワークの解明」第1回研究会
  • Place of Presentation: 国際高等研究所(京都)
  • Year and Date: 2013-04-21
• [Presentation] プロテアソーム集合因子Nas2 を介したプロテアソームの分子集合機構の構造基盤 (2013)
  • Author(s): 佐藤匡史、植草義徳、廣本武史、矢木真穂、矢木宏和、水島恒裕、佐伯泰、田中啓二、加藤晃一
  • Organizer: 日本薬学会 第133回年会
  • Place of Presentation: パシフィコ横浜(神奈川)
  • Year and Date: 2013-03-29
• [Presentation] NMR とSANS によるタンパク質の動的構造解析 (2013)
  • Author(s): 加藤晃一
  • Organizer: 中性子連携研究会
  • Place of Presentation: 京都大学東京オフィス(東京)
  • Year and Date: 2013-03-13
• [Presentation] プロテアソームを構成するサブユニットとその集合因子Ump1p の相互作用解析 (2013)
  • Author(s): 大川慶祐、矢木宏和、植草義徳、矢木真穂、水島恒裕、佐藤匡史、佐伯泰、田中啓二、加藤晃一
  • Organizer: 日本生物物理学会中部支部講演会
  • Place of Presentation: 名古屋大学(愛知)
  • Year and Date: 2013-02-19
• [Presentation] 生命分子複合体の動態解明への実験的アプローチ (2013)
  • Author(s): 加藤晃一
  • Organizer: 京都大学原子炉実験所 粒子線基礎物性研究特別講演会
  • Place of Presentation: 京都大学原子炉実験所(大阪)
  • Year and Date: 2013-02-12
• [Presentation] ATP 非依存性シャペロンの構造ダイナミクスと機能発現メカニズム (2012)
  • Author(s): 加藤晃一
  • Organizer: 新学術領域「揺らぎと生体機能」「水和とATP」合同公開シンポジウム「ゆらぎと水-生命のエネルギーと分子機構を探る」
  • Place of Presentation: 大阪ガーデンパレス(大阪)
  • Year and Date: 2012-09-14
• [Presentation] 重水素化標識を用いたタンパク質複合体の4次構造ダイナミクスの研究 (2012)
  • Author(s): 加藤晃一、杉山正明
  • Organizer: J-PARC プロジェクト課題「生体高分子の構造-ダイナミクス相関の解析」キックオフ会議
  • Place of Presentation: J-PARCセンター(茨城)
  • Year and Date: 2012-07-19
• [Presentation] プロテアソーム19S 制御因子複合体形成シャペロンHsm3 の構造解析 (2012)
  • Author(s): 高木賢治、金相佑、加藤晃一、田中啓二、佐伯泰、水島恒裕
  • Organizer: 第12回日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場(愛知)
  • Year and Date: 2012-06-20
• [Presentation] 新規プロテアソーム活性化因子PbaB の構造と機能 (2012)
  • Author(s): 雲井健太郎、佐藤匡史、矢木宏和、廣本武史、水島恒裕、内山進、野田勝紀、村田和義、神谷由紀子、加藤晃一
  • Organizer: 第12回日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場(愛知)
  • Year and Date: 2012-06-20
• [Presentation] 古細菌タンパク質PbaB によるATP 非依存的プロテアソーム活性化機構の構造基盤 (2012)
  • Author(s): 雲井健太郎、佐藤匡史、廣本武史、水島恒裕、神谷由紀子、野田勝紀、内山進、村田和義、矢木宏和、加藤晃一
  • Organizer: 第76回日本生化学会 中部支部例会・シンポジウム
  • Place of Presentation: 岡崎コンファレンスセンター(愛知)
  • Year and Date: 2012-05-26