Mechanism of the proteasome nuclear localization

• Project/Area Number: 22657037
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Functional biochemistry
• Research Institution: 財団法人東京都医学総合研究所 (2011); Tokyo Metropolitan Organization for Medical Research (2010)
• Principal Investigator: SAEKI Yasushi 財団法人東京都医学総合研究所, 生体分子先端研究分野, 主席研究員 (80462779)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,420,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2010: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: 細胞内タンパク質分解 / 蛋白質分解 / プロテアソーム / 細胞質核間輸送担体 / 酵母 / 微生物

Research Abstract

The 26S proteasome is a highly organized protease complex that is responsible for targeted protein degradation in both the cytosolic and nuclear compartment. In budding yeast and several cancer cells, it is known that the 26S proteasome is mainly localized in the nucleus. Although several studies have described the mechanism of the nuclear translocation, it is still unclear when the proteasomes enter the nucleus upon their assembly process. Using dual color fluorescence cross correlation spectrometry(FCCS), we determined local concentration and dynamics of the 26S proteasome in living yeast cells. Surprisingly, almost all the proteasome subunits were incorporated into the 26S proteasome in both the cytosol and nucleus. Importantly, the proteasome dynamics was not changed in the importin.(srp1-49) mutant cells in which the 26S proteasome retains the cytosol. These results suggest that the 26S proteasome is fully assembled in the cytoplasm prior to the nuclear translocation.

Research Products

• [Journal Article] Proteasomal degradation resolves competition between cell polarization and cellular wound healing (2012)
  • Author(s): Kono, K., Saeki, Y., Yoshida, S., Tanaka, K., and Pellman, D.
  • Journal Title: Cell Volume: (in press)
  • Peer Reviewed
• [Journal Article] Structural basis for specific recognition of Rpt1p, an ATPase subunit of the 26S proteasome, by the proteasome-dedicated chaperone Hsm3p (2012)
  • Author(s): Takagi, K., Kim, S., Yukii, H., Ueno, M., Morishita, R., Endo, Y., Kato, K., Tanaka, K., Saeki, Y., and Mizushima, Y.
  • Journal Title: J Biol Chem Volume: 287 Pages: 12172-12182
  • Peer Reviewed
• [Journal Article] Localization of the proteasomal ubiquitin receptors Rpn10 and Rpn13 by electron cryomicroscopy (2012)
  • Author(s): Sakata, E., Bohn, S., Mihalache, O., Kiss, P., Beck, F., Nagy, I., Nickell, S., Tanaka, K., Saeki, Y., Forster, F., and Baumeister, W.
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Pages: 1479-1484
  • Peer Reviewed
• [Journal Article] The proteasome : molecular machinery and pathophysiological roles (2012)
  • Author(s): Tanaka, K., Mizushima, T., and Saeki, Y.
  • Journal Title: Biol Chem Volume: 393 Pages: 217-234
  • Peer Reviewed
• [Journal Article] Assembly and function of the proteasome (2012)
  • Author(s): Saeki, Y and Tanaka, K.
  • Journal Title: Methods Mol Biol Volume: 832 Pages: 315-337
  • Peer Reviewed
• [Journal Article] Structural basis for specific recognition of Rpt1p, an ATPase subunit of the 26S proteasome, by a proteasome-dedicated chaperone Hsm3p (2012)
  • Author(s): Takagi, T., et al
  • Journal Title: J Biol Chem Volume: 287 Pages: 12172-12182
  • Peer Reviewed
• [Journal Article] The catalytic activity of Ubp6 enhances maturation of the proteasomal regulatory particle (2011)
  • Author(s): Sakata, E., Stengel, F., Fukunaga, K., Zhou, M., Saeki, Y., Forster, F., Baumeister, W., Tanaka, K., and Robinson, CV.
  • Journal Title: Mol Cell Volume: 42 Pages: 637-649
  • Peer Reviewed
• [Journal Article] Sharpin is a component of the NF-κB activating linear ubiquitin chain assembly complex (2011)
  • Author(s): Tokunaga, F., Nakagawa, T., Nakahara1, M., Saeki, Y., Taniguchi, M., Sakata, S., Tanaka, K., Nakano, H., and Iwai, I.
  • Journal Title: Nature Volume: 471 Pages: 633-636
  • Peer Reviewed
• [Journal Article] Sharpin is a component of the NF-κB activating linear ubiquitin chain assembly complex. (2011)
  • Author(s): Tokunaga, F., et al.
  • Journal Title: Nature Volume: 471 Pages: 633-636
  • Peer Reviewed
• [Journal Article] The catalytic activity of Ubp6 enhances maturation of the proteasomal regulatory particle. (2011)
  • Author(s): Sakata, E., et al.
  • Journal Title: Mol.Cell Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] Crystal structure of yeast Rpn14, a chaperone of the 19S regulatory particles of the proteasome (2010)
  • Author(s): Kim, S., Saeki, Y., Fukunaga, K., Suzuki, A., Takagi, K., Yamane, T., Tanaka, K., Mizushima, T., and Kato, K.
  • Journal Title: J Biol Chem Volume: 285 Pages: 15159-151566
  • Peer Reviewed
• [Journal Article] Dissection of the assembly pathway of the proteasome lid in Saccharomyces cerevisiae (2010)
  • Author(s): Fukunaga, K., Kudo, T., Toh-e, A., Tanaka, K., and Saeki, Y.
  • Journal Title: Biochem Biophys Res Commun Volume: 396 Pages: 1048-1053
  • Peer Reviewed
• [Journal Article] Polyubiquitin conjugation to NEMO by triparite motif protein 23(TRIM23) is critical in antiviral defense (2010)
  • Author(s): Arimoto, K. I., Funami, K., Saeki, Y., Tanaka, K., Okawa, K., Takeuchi, O., Akira, S., Murakami, Y., and Shimotohno, K.
  • Journal Title: Proc Natl Acad Sci USA Volume: 107 Pages: 15856-15861
  • Peer Reviewed
• [Journal Article] Crystal structure of yeast Rpn14, a chaperone of the 19S regulatory particle of the proteasome. (2010)
  • Author(s): Kim, S., et al.
  • Journal Title: J.Biol.Chem. Volume: 285 Pages: 15159-15166
  • Peer Reviewed
• [Journal Article] Dissection of the assembly pathway of the proteasome lid in Saceharomyces cerevisiae. (2010)
  • Author(s): Fukunaga, K., et al.
  • Journal Title: Biochem.Biophys.Res.Commun. Volume: 396 Pages: 1048-1053
  • Peer Reviewed
• [Journal Article] Polyubiquitin conjugation to NEMO by tripartite motif protein 23 (TRIM23) is critical in antiviral defense. (2010)
  • Author(s): Arimoto, KI, et al.
  • Journal Title: Proc.Natl.Acad.Sci.USA. Volume: 107 Pages: 15856-15861
  • Peer Reviewed
• [Presentation] 出芽酵母26Sプロテアソームは細胞質で完成する (2011)
  • Author(s): 佐伯泰、白燦基、東江昭夫、田中啓二
  • Organizer: 第44回日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2011-12-15
• [Presentation] (2011)
  • Author(s): 佐伯泰
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 2011-12-15
• [Presentation] 東京大学大学院農学生命科学科 (2011)
  • Author(s): 佐伯泰
  • Organizer: 第7回応用動物科学セミナー「プロテアソーム研究の最前線」
  • Place of Presentation: 東京
  • Year and Date: 2011-10-25
• [Presentation] Rad23とUbp6は26Sプロテアソームの分子集合に関与する (2011)
  • Author(s): 佐伯泰、福永圭佑、坂田絵里、Florian Stengel、Carol V. Robinson、Wolfgang Baumeister、田中啓二
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2011-09-24
• [Presentation] 出芽酵母26Sプロテアソームは細胞質で完成する (2011)
  • Author(s): 佐伯泰、白燦基、東江昭夫、田中啓二
  • Organizer: 酵母遺伝学フォーラム第44回研究報告会
  • Place of Presentation: 博多
  • Year and Date: 2011-09-07
• [Presentation] 26Sプロテアソームの分子集合と作動原理 (2011)
  • Author(s): 佐伯泰
  • Organizer: 第19回酵母合同シンポジウム
  • Place of Presentation: 東京(招待講演)
  • Year and Date: 2011-06-24
• [Presentation] 26Sプロテアソームの分子集合機構 (2011)
  • Author(s): 佐伯泰,福永圭佑,工藤泰,川村ひとみ,田中啓二
  • Organizer: 平成22年度ターゲットタンパク研究プログラム成果発表会
  • Place of Presentation: 東京
  • Year and Date: 2011-03-11
• [Presentation] Assembly, structure, and function of the 26S proteasome. (2011)
  • Author(s): Yasushi Saeki
  • Organizer: Korea-Japan Symposium on Protein Metabolism
  • Place of Presentation: Seoul, Korea(招待講演)
  • Year and Date: 2011-03-11
• [Presentation] The 26S proteasome completes its assembly process in the cytoplasm prior to the nuclear translocation (2011)
  • Author(s): Haruka Yukii, Yasushi Saeki, Changi Pack, Keisuke Fukunaga, Eri Sakata, Yasushi Sako, Akio Toh-e, Wolfgang Baumeister, Keiji Tanaka
  • Organizer: EMBO Conference, Ubiquitin and ubiquitin-like modifiers
  • Place of Presentation: Cavtat, Croatia
• [Presentation] Assembly, structure, and function of the 26S proteasome (2011)
  • Author(s): Yasushi Saeki
  • Organizer: Korea-Japan Symposium on Protein Metabolism
  • Place of Presentation: Seoul, Korea
• [Presentation] 26Sプロテアソームはどこで完成するか? (2010)
  • Author(s): 佐伯泰, 白燦基, 川村ひとみ, 東江昭夫, 佐甲靖志, 田中啓二
  • Organizer: BMB2010(第33回分子生物学会、第83回日本生化学会合同年会)
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-09
• [Presentation] 26Sプロテアソームはどこで完成するか (2010)
  • Author(s): 佐伯泰,白燦基,川村ひとみ,東江昭夫,佐甲靖志,田中啓二
  • Organizer: BMB2010(第33回分子生物学会、第83回日本生化学会合同年会)
  • Place of Presentation: 神戸
• [Presentation] プロテアソーム19S制御因子複合体の分子集合機構 (2010)
  • Author(s): 佐伯泰,福永圭佑,田中啓二
  • Organizer: 平成22年度ターゲットタンパク研究プログラム成果発表会
  • Place of Presentation: 東京
• [Presentation] 26Sプロテアソームの分子集合と作動原理 (2010)
  • Author(s): 佐伯泰
  • Organizer: 第19回酵母合同シンポジウム
  • Place of Presentation: 東京
• [Book] Ubiquitin Family Modifiers and the Proteasome (2012)
  • Author(s): Saeki, Y., Tanaka, K.
  • Total Pages: 22
  • Publisher: Humana Press
• [Book] タンパク質分解研究による生態防御 (2012)
  • Author(s): 佐伯泰
  • Total Pages: 6
  • Publisher: 羊土社
• [Book] みえてきた26Sプロテアソームの形成機構の全貌,「細胞内分解系によるリノベーションの分子機構に迫る」実験医学 (2011)
  • Author(s): 佐伯泰,福永圭佑
  • Publisher: 羊土社
• [Book] 脱ユビキチン化酵素Ubp6はプロテアソームの分子集合を制御する.ライフサイエンス「新着論文レビュー」 (2011)
  • Author(s): 佐伯泰,田中啓二
  • URL: http://first.lifesciencedb.jp/archives/3121
• [Book] Methods in Molecular Biology (Ubiquitin family modifiers and the proteasome) (2011)
  • Author(s): Saeki, Y., Tanaka, K.
  • Publisher: Humana Press 印刷中
• [Book] 実験医学・増刊号(細胞内分解系によるリノベーションの分子機構に迫る) (2011)
  • Author(s): 佐伯泰、福永圭佑
  • Publisher: 羊土社 印刷中
• [Book] 日本臨牀 特集 分子標的薬治療 (2010)
  • Author(s): 佐伯泰、福永圭佑、田中啓二
  • Total Pages: 5
  • Publisher: 日本臨牀社
• [Remarks]
  • URL: http://www.igakuken.or.jp/pro-meta/
• [Remarks]
  • URL: http://check.shonai/pro-meta/studies/saeki.html
• [Remarks]
  • URL: http://www.rinshoken.or.jp/MO/