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2018 Fiscal Year Annual Research Report

Global analyses of nascent chain folding and chaperone effects

Planned Research

Project AreaNascent-chain Biology
Project/Area Number 26116002
Research InstitutionTokyo Institute of Technology

Principal Investigator

田口 英樹  東京工業大学, 科学技術創成研究院, 教授 (40272710)

Co-Investigator(Kenkyū-buntansha) 今高 寛晃  兵庫県立大学, 工学研究科, 教授 (50201942)
富田 野乃 (竹内野乃)  東京大学, 大学院新領域創成科学研究科, 准教授 (80323450)
Project Period (FY) 2014-07-10 – 2019-03-31
Keywords新生鎖 / フォールディング / シャペロン / リボソーム / 無細胞翻訳系
Outline of Annual Research Achievements

本計画研究では、翻訳途上の新生ポリペプチド鎖(新生鎖)がどのような過程を経てできあがるのかを生化学、分子生物学、遺伝学、生物物理学などさまざまな手法を駆使して解明することを目的とした。大腸菌、出芽酵母、ヒト細胞由来の翻訳系の再構成系を中心に研究を推進し、最終年度はこれまでの成果の論文発表に加え、本計画研究が発端として見いだされた新たな知見の発展研究を行った。具体的には以下のような成果を得た。
【田口】1)翻訳に共役したタンパク質フォールディングを大腸菌の再構築型無細胞翻訳系とプロテアーゼを用いて行う系を確立し、100種類弱のタンパク質のフォールディングを解析した(Niwa T et al Protein Sci 2019)。2)出芽酵母の500種類程度のタンパク質の翻訳に共役したフォールディングを解析した(Uemura E et al Sci Rep 2018)。3)大腸菌抽出液に含まれるタンパク質の相分離のしやすさをプロテオームレベルで解析した(Nojima T et al Biomacromolecules 2018)。4)天然変性タンパク質の動態を高速原子間力顕微鏡で観察し、構造形成のモデルを提案した(Konno H et al PNAS 2020)。
【今高】ヒト因子由来再構成翻訳系を使うことでウイルスにおける非典型的な翻訳現象(HCV IRESによる翻訳開始現象)の分子機構を解明した(Yokoyama T et al Mol Cell 2019)。
【富田】出芽酵母由来の再構築型翻訳系を完成させて、論文発表を行った(Abe T et al J Biochem 2020a)。さらに、完成した再構築型翻訳系を使って、翻訳伸長因子eIF5などの機能解析を行った(Abe T et al J Biochem 2020b)。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

  • Research Products

    (24 results)

All 2020 2019 2018 Other

All Int'l Joint Research (3 results) Journal Article (15 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 15 results,  Open Access: 4 results) Presentation (4 results) (of which Int'l Joint Research: 4 results,  Invited: 3 results) Book (2 results)

  • [Int'l Joint Research] コロラド州立大学(米国)

    • Country Name
      U.S.A.
    • Counterpart Institution
      コロラド州立大学
  • [Int'l Joint Research] ユトレヒト大学(オランダ)

    • Country Name
      NETHERLANDS
    • Counterpart Institution
      ユトレヒト大学
  • [Int'l Joint Research] CSIR-IMTEC(インド)

    • Country Name
      INDIA
    • Counterpart Institution
      CSIR-IMTEC
  • [Journal Article] Nascent SecM chain interacts with outer ribosomal surface to stabilize translation arrest2020

    • Author(s)
      Muta Mikihisa、Iizuka Ryo、Niwa Tatsuya、Guo Yuanfang、Taguchi Hideki、Funatsu Takashi
    • Journal Title

      Biochemical Journal

      Volume: 477 Pages: 557~566

    • DOI

      doi: 10.1042/BCJ20190723

    • Peer Reviewed / Open Access
  • [Journal Article] Dynamics of oligomer and amyloid fibril formation by yeast prion Sup35 observed by high-speed atomic force microscopy2020

    • Author(s)
      Konno Hiroki、Watanabe-Nakayama Takahiro、Uchihashi Takayuki、Okuda Momoko、Zhu Liwen、Kodera Noriyuki、Kikuchi Yousuke、Ando Toshio、Taguchi Hideki
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 117 Pages: 7831~7836

    • DOI

      doi: 10.1073/pnas.1916452117

    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Translation efficiency affects the sequence-independent +1 ribosomal frameshifting by polyamines2020

    • Author(s)
      Oguro Akihiro、Shigeta Tomoaki、Machida Kodai、Suzuki Tomoaki、Iwamoto Takeo、Matsufuji Senya、Imataka Hiroaki
    • Journal Title

      The Journal of Biochemistry

      Volume: . Pages: in press

    • DOI

      doi: 10.1093/jb/mvaa032

    • Peer Reviewed
  • [Journal Article] Reconstitution of yeast translation elongation and termination in vitro utilizing CrPV IRES-containing mRNA2020

    • Author(s)
      Abe Taisho、Nagai Riku、Imataka Hiroaki、Takeuchi-Tomita Nono
    • Journal Title

      The Journal of Biochemistry

      Volume: 167 Pages: 441~450

    • DOI

      doi: 10.1093/jb/mvaa021

    • Peer Reviewed
  • [Journal Article] In vitro yeast reconstituted translation system reveals function of eIF5A for synthesis of long polypeptide2020

    • Author(s)
      Abe Taisho、Nagai Riku、Shimazaki Shunta、Kondo Shunta、Nishimura Satoshi、Sakaguchi Yuriko、Suzuki Tsutomu、Imataka Hiroaki、Tomita Kozo、Takeuchi-Tomita Nono
    • Journal Title

      The Journal of Biochemistry

      Volume: 167 Pages: 451~462

    • DOI

      doi: 10.1093/jb/mvaa022

    • Peer Reviewed
  • [Journal Article] Disruption of the Gene trx-m1 Impedes the Growth of Anabaena sp. PCC 7120 under Nitrogen Starvation2019

    • Author(s)
      Deschoenmaeker Frederic、Mihara Shoko、Niwa Tatsuya、Taguchi Hideki、Nomata Jiro、Wakabayashi Ken-Ichi、Hisabori Toru
    • Journal Title

      Plant and Cell Physiology

      Volume: 60 Pages: 1504~1513

    • DOI

      doi: 10.1093/pcp/pcz056

    • Peer Reviewed
  • [Journal Article] Translation‐coupled protein folding assay using a protease to monitor the folding status2019

    • Author(s)
      Niwa Tatsuya、Uemura Eri、Matsuno Yuki、Taguchi Hideki
    • Journal Title

      Protein Science

      Volume: 28 Pages: 1252-1261

    • DOI

      doi: 10.1002/pro.3624

    • Peer Reviewed
  • [Journal Article] Large-scale analysis of diffusional dynamics of proteins in living yeast cells using fluorescence correlation spectroscopy2019

    • Author(s)
      Fukuda Takafumi、Kawai-Noma Shigeko、Pack Chan-Gi、Taguchi Hideki
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 520 Pages: 237~242

    • DOI

      doi: 10.1016/j.bbrc.2019.09.066

    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Catalyst-proximity protein chemical labelling on affinity beads targeting endogenous lectins2019

    • Author(s)
      Tsushima Michihiko、Sato Shinichi、Niwa Tatsuya、Taguchi Hideki、Nakamura Hiroyuki
    • Journal Title

      Chemical Communications

      Volume: 55 Pages: 13275~13278

    • DOI

      doi: 10.1039/c9cc05231c

    • Peer Reviewed
  • [Journal Article] HCV IRES Captures an Actively Translating 80S Ribosome2019

    • Author(s)
      Yokoyama Takeshi、Machida Kodai、Iwasaki Wakana、Shigeta Tomoaki、Nishimoto Madoka、Takahashi Mari、Sakamoto Ayako、Yonemochi Mayumi、Harada Yoshie、Shigematsu Hideki、Shirouzu Mikako、Tadakuma Hisashi、Imataka Hiroaki、Ito Takuhiro
    • Journal Title

      Molecular Cell

      Volume: 74 Pages: 1205~1214.e8

    • DOI

      doi: 10.1016/j.molcel.2019.04.022

    • Peer Reviewed
  • [Journal Article] Large-scale aggregation analysis of eukaryotic proteins reveals an involvement of intrinsically disordered regions in protein folding2018

    • Author(s)
      Uemura Eri、Niwa Tatsuya、Minami Shintaro、Takemoto Kazuhiro、Fukuchi Satoshi、Machida Kodai、Imataka Hiroaki、Ueda Takuya、Ota Motonori、Taguchi Hideki
    • Journal Title

      Scientific Reports

      Volume: 8 Pages: 678

    • DOI

      doi: 10.1038/s41598-017-18977-5

    • Peer Reviewed / Open Access
  • [Journal Article] The Absence of Thioredoxin m1 and Thioredoxin C in Anabaena sp. PCC 7120 Leads to Oxidative Stress2018

    • Author(s)
      Deschoenmaeker Frederic、Mihara Shoko、Niwa Tatsuya、Taguchi Hideki、Wakabayashi Ken-Ichi、Hisabori Toru
    • Journal Title

      Plant and Cell Physiology

      Volume: 59 Pages: 2432-2441

    • DOI

      doi: 10.1093/pcp/pcy163

    • Peer Reviewed
  • [Journal Article] Electrostatic interactions between middle domain motif-1 and the AAA1 module of the bacterial ClpB chaperone are essential for protein disaggregation2018

    • Author(s)
      Sugita Saori、Watanabe Kumiko、Hashimoto Kana、Niwa Tatsuya、Uemura Eri、Taguchi Hideki、Watanabe Yo-hei
    • Journal Title

      Journal of Biological Chemistry

      Volume: 293 Pages: 19228~19239

    • DOI

      doi: 10.1074/jbc.RA118.005496

    • Peer Reviewed / Open Access
  • [Journal Article] Proteome Analysis of Phase-Separated Condensed Proteins with Ionic Surfactants Revealed Versatile Formation of Artificial Biomolecular Condensates2018

    • Author(s)
      Nojima Tatsuya、Niwa Tatsuya、Taguchi Hideki
    • Journal Title

      Biomacromolecules

      Volume: 20 Pages: 539~545

    • DOI

      doi: 10.1021/acs.biomac.8b01379

    • Peer Reviewed
  • [Journal Article] Dynamic interaction of poly(A)-binding protein with the ribosome2018

    • Author(s)
      Machida Kodai、Shigeta Tomoaki、Yamamoto Yuki、Ito Takuhiro、Svitkin Yuri、Sonenberg Nahum、Imataka Hiroaki
    • Journal Title

      Scientific Reports

      Volume: 8 Pages: 17435

    • DOI

      doi: 10.1038/s41598-018-35753-1

    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Nascent chain-induced ribosome dynamics regulation2019

    • Author(s)
      Hideki Taguchi
    • Organizer
      EMBO Workshop on Protein quality control: From mechanisms to disease, April 28-May 3, 2019, Mallorca, Spain
    • Int'l Joint Research / Invited
  • [Presentation] Conversion of a chaperonin GroEL-dependent substrate protein into GroEL-independent folders2018

    • Author(s)
      Hideki Taguchi
    • Organizer
      Conference on Conformational Transitions in Proteins
    • Int'l Joint Research / Invited
  • [Presentation] Intrinsic ribosome destabilization underlies translation and provides an organism with a strategy of environmental sensing2018

    • Author(s)
      Hideki Taguchi
    • Organizer
      Cold Spring Harbor Laboratory meeting on Protein Homeostasis in Health and Diseases., April 17-21, 2018, Cold Spring Harbor, USA,
    • Int'l Joint Research
  • [Presentation] Nascent chain-induced ribosome dynamics regulation2018

    • Author(s)
      Hideki Taguchi
    • Organizer
      International Symposium on Proteins: From the Cradle to the Grave, August 26-29, 2018, Enryakuji, Japan
    • Int'l Joint Research / Invited
  • [Book] 池上彰が聞いてわかった生命のしくみ 東工大で生命科学を学ぶ2020

    • Author(s)
      池上 彰、岩崎博史、田口 英樹
    • Total Pages
      352
    • Publisher
      朝日新聞出版
    • ISBN
      4022620102
  • [Book] 実験医学2019年11月号 特集「再定義されるタンパク質の常識」2019

    • Author(s)
      田口英樹
    • Total Pages
      147
    • Publisher
      羊土社
    • ISBN
      4758125252

URL: 

Published: 2021-01-27  

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