Construction of quantitative reporter system for branched ubiquitin chains

• Project/Area Number: 16K14702
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: OHTAKE Fumiaki 公益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 (60447373)
• Co-Investigator(Kenkyū-buntansha): 安彦 行人 国立医薬品食品衛生研究所, 毒性部, 主任研究官 (40370944)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: ユビキチン / 翻訳後修飾 / タンパク質 / シグナル伝達 / 質量分析 / 蛋白質 / 遺伝子

Outline of Final Research Achievements

Ubiquitin chains of different linkage types regulate diverse biological pathways.However,functions of branched ubiquitin chains had been largely unknown. We established reporter systems to quantify K48/K63 branched ubiquitin chains in yeast and in mice. Moreover, we screened and identified regulators of branched ubiquitin chains in yeast.

Academic Significance and Societal Importance of the Research Achievements

ユビキチン鎖は分岐によって複雑な高次構造を形成しうることが近年明らかにされてきましたが、分岐型ユビキチン鎖の定量は技術的に困難でした。本研究では分岐型ユビキチン鎖を出芽酵母および動物個体において定量するためのレポーター系の構築に成功したことから、分岐型ユビキチン鎖の形成・制御機構の有用な解析系となることが期待されます。

Research Products

• [Journal Article] Methods to measure ubiquitin chain length and linkage. (2019)
  • Author(s): Ohtake, F., Tsuchiya, H., Tanaka, K., and Saeki, Y.
  • Journal Title: Methods Enzymol Volume: 618 Pages: 105-133
  • DOI: 10.1016/bs.mie.2018.12.019
  • Peer Reviewed / Open Access
• [Journal Article] K63 ubiquitylation triggers proteasomal degradation by seeding branched ubiquitin chains. (2018)
  • Author(s): Ohtake, F., Tsuchiya, H., Saeki, Y., and Tanaka, K
  • Journal Title: PNAS Volume: 115 Issue: 7 Pages: 1401-1408
  • DOI: 10.1073/pnas.1716673115
  • Peer Reviewed
• [Journal Article] Ub-ProT reveals global length and composition of protein ubiquitylation in cells (2018)
  • Author(s): Tsuchiya, H., Burana, D., Ohtake, F., Arai, N., Kaiho, A., Komada, M., Tanaka,K., and Saeki, Y.
  • Journal Title: Nature Commun Volume: 9 Issue: 1 Pages: 524-524
  • DOI: 10.1038/s41467-018-02869-x
  • Peer Reviewed / Open Access
• [Journal Article] The K48-K63 branched ubiquitin chain regulates NF-kappaB signaling (2016)
  • Author(s): Ohtake F, Saeki Y, Ishido S, Kanno J, Tanaka K
  • Journal Title: Mol. Cell Volume: 64 Issue: 2 Pages: 251-266
  • DOI: 10.1016/j.molcel.2016.09.014
  • Peer Reviewed
• [Journal Article] The complexity of ubiquitin architecture. (2016)
  • Author(s): Ohtake, F., and Tsuchiya, H
  • Journal Title: J Biochem Volume: 161 Pages: 125-133
  • DOI: 10.1093/jb/mvw088
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] The ubiquitin ligases ITCH and UBR5 regulate proteasomal degradation through K48/K63 branched ubiquitin chains (2018)
  • Author(s): Ohtake F, Saeki Y, Tanaka K.
  • Organizer: 第41回日本分子生物学会シンポジウム
  • Invited
• [Presentation] Post-translational modification code of ubiquitin in signal transduction and protein degradation (2018)
  • Author(s): Ohtake F, Saeki Y, Tanaka K.
  • Organizer: 第91回日本生化学会シンポジウム
  • Invited
• [Presentation] The NEDD4 family ubiquitin ligase ITCH regulates proteasomal degradation by seeding branched ubiquitin chains. (2018)
  • Author(s): 5.Ohtake F, Saeki Y, Tanaka K
  • Organizer: FASEB Conference「Ubiquitin and cellular regulation」
  • Int'l Joint Research
• [Presentation] 炎症応答を制御する新たなユビキチン・シグナル (2017)
  • Author(s): 大竹史明、田中啓二
  • Organizer: 第44回日本毒性学会
  • Invited
• [Presentation] 出芽酵母におけるK48/K63分岐型ポリユビキチン鎖の解析 (2017)
  • Author(s): 河野有紗、大竹史明、佐伯泰、田中啓二
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] Role of the K48/K63 branched ubiquitin chains in signal transduction and proteasomal degradation (2017)
  • Author(s): Ohtake F., Tanaka K
  • Organizer: EMBO conference「Ubiquitin and SUMO」
  • Int'l Joint Research
• [Presentation] Cellular function of the Lys48/Lys63 branched ubiquitin chain. (2016)
  • Author(s): Ohtake F
  • Organizer: 国際シンポジウム「Diverse functions of Ubiquitin : Degradation, Signaling, and Beyond」
  • Place of Presentation: 京都大学（京都府京都市）
  • Int'l Joint Research / Invited
• [Presentation] Quantitative analysis of the branched ubiquitin chains (2016)
  • Author(s): Ohtake F, Saeki Y, Tanaka K
  • Organizer: 第39回日本分子生物学会シンポジウム
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Invited
• [Remarks] 蛋白質代謝研究室ホームページ
  • URL: http://www.igakuken.or.jp/pro-meta/