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In vitro translation and analysis of neurodegenerative diseases

Research Project

Project/Area Number 17H04995
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field System genome science
Research InstitutionUniversity of Hyogo

Principal Investigator

Machida Kodai  兵庫県立大学, 工学研究科, 准教授 (20553093)

Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥23,270,000 (Direct Cost: ¥17,900,000、Indirect Cost: ¥5,370,000)
Fiscal Year 2020: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2018: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2017: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
Keywords再構成型タンパク質合成系 / 神経変性疾患 / 翻訳機構 / バイオテクノロジー / 生体分子 / 脳神経疾患
Outline of Final Research Achievements

We succeeded in reproducing Repeat Associated Non-AUG translation, which is an irregular protein translation depends on CAG repeat sequences that cause human neurodegenerative diseases using a reconstituted protein synthesis system constructed with purified human translation factors. These results indicating that the translation reactions that cause neurodegenerative diseases occur if there is a translation-related factors such as mRNA, ribosome, translation initiation factors, translation elongation factors, and translation termination factors. This study established the technical basis for reproducing and analyzing the irregular translation mechanism that causes neurodegenerative diseases in vitro.

Academic Significance and Societal Importance of the Research Achievements

本研究では、神経変性疾患の原因となるタンパク質凝集、具体的にはポリグルタミン病を引き起こすCAGリピート配列からの翻訳と翻訳産物の凝集をヒト因子由来の試験管内翻訳系で再現・解析するための技術基盤を確立した。リポソーム化を取り入れ細胞サイズの反応系場での解析も行えるようにした。本実験系はヒトの翻訳関連因子のみで再構築したものであり、試験管内でヒトの疾患に関連する遺伝子の一連の翻訳反応について新たな知見を得ることができるようになったため学術的意義が大きい。また、ヒト因子由来の実験系であるため、得られたデータを直接的に治療法開発へ応用できる可能性が高いという利点もあり、社会的な意義も大きい。

Report

(5 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (30 results)

All 2021 2020 2019 2018 2017

All Journal Article (8 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 8 results,  Open Access: 6 results) Presentation (22 results) (of which Int'l Joint Research: 2 results,  Invited: 3 results)

  • [Journal Article] Distinct roles and actions of protein disulfide isomerase family enzymes in catalysis of nascent-chain disulfide bond formation2021

    • Author(s)
      Hirayama, C., Machida, K., Noi, K., Murakawa, T., Okumura, M., Ogura, T., Imataka, H., and Inaba, K.*
    • Journal Title

      iScience

      Volume: 4 Issue: 4 Pages: 102296-102296

    • DOI

      10.1016/j.isci.2021.102296

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] eIF4G-driven translation initiation of downstream ORFs in mammalian cells2020

    • Author(s)
      Nobuta Risa、Machida Kodai、Sato Misaki、Hashimoto Satoshi、Toriumi Yasuhito、Nakajima Shizuka、Suto Daiki、Imataka Hiroaki、Inada Toshifumi
    • Journal Title

      Nucleic Acids Research

      Volume: 48 Issue: 18 Pages: 10441-10455

    • DOI

      10.1093/nar/gkaa728

    • Related Report
      2020 Annual Research Report
    • 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: 印刷中 Issue: 6 Pages: 1205-1214

    • DOI

      10.1016/j.molcel.2019.04.022

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [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 Issue: 1 Pages: 17435-17435

    • DOI

      10.1038/s41598-018-35753-1

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Palm-sized Ag ion emission gun operated at roomtemperature in non-vacuum atmosphere2018

    • Author(s)
      Yusuke Daiko, Kyohei Segawa, Kodai Machida, Hiroaki Imataka, Sawao Honda, and Yuji Iwamoto
    • Journal Title

      Adv. Engneering Mater.

      Volume: 20 Issue: 9 Pages: 1800198-1800198

    • DOI

      10.1002/adem.201800198

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Huntingtin polyglutamine-dependent protein aggregation in reconstituted cells.2018

    • Author(s)
      Kodai Machida, Kuru Kanzawa, Tomoaki Shigeta, Yuki Yamamoto, Kanta Tsumoto, and Hiroaki Imataka
    • Journal Title

      ACS synthetic Biology

      Volume: 7 Issue: 2 Pages: 377-383

    • DOI

      10.1021/acssynbio.7b00372

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Large-scale aggregation analysis of eukaryotic proteins reveals an involvement of intrinsically disordered regions in protein folding.2018

    • Author(s)
      Uemura E, Niwa T, Minami S, Takemoto K, Fukuchi S, Machida K, Imataka H, Ueda T, Ota M, Taguchi H.
    • Journal Title

      Sci Rep.

      Volume: 8 Issue: 1 Pages: 678-678

    • DOI

      10.1038/s41598-017-18977-5

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] H+ emission under room temperature and non-vacuum atmosphere from a sol?gel-derived nanoporous emitter2017

    • Author(s)
      Daiko Yusuke、Mizutani Satoshi、Machida Kodai、Imataka Hiroaki、Honda Sawao、Iwamoto Yuji
    • Journal Title

      Journal of Sol-Gel Science and Technology

      Volume: 83 Issue: 2 Pages: 252-258

    • DOI

      10.1007/s10971-017-4430-z

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Presentation] 試験管内翻訳系:合成生物学および創薬への応用2020

    • Author(s)
      今高寛晃、町田幸大
    • Organizer
      第43回 日本分子生物学会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] ヒト由来無細胞系を用いたアクチン新生鎖のフォールディング解析2020

    • Author(s)
      白子太紀、町田幸大、今高寛晃
    • Organizer
      第43回 日本分子生物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 無細胞タンパク質翻訳系を用いたRAN翻訳の解析2020

    • Author(s)
      鈴木智明、小黒明広、町田幸大、今高寛晃
    • Organizer
      第43回 日本分子生物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] In vitro actin biogenesis using a human reconstituted translation/folding coupled system2020

    • Author(s)
      宮脇翔馬、町田幸大、今高寛晃
    • Organizer
      第43回 日本分子生物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 翻訳開始因子eIF4BおよびeIF4HによるIRES依存的翻訳開始機構の制御2020

    • Author(s)
      友廣 拓生、深尾 亜喜良、町田 幸大、船上 仁範、今高 寛晃、藤原 俊伸
    • Organizer
      第43回 日本分子生物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] アクチン生合成の試験管内再現2019

    • Author(s)
      町田幸大、神澤空流、白子太紀、井寄真仁、今高寛晃
    • Organizer
      第19回 日本蛋白質科学会年会・第71回日本細胞生物学会大会 合同年次大
    • Related Report
      2019 Annual Research Report
  • [Presentation] 試験管内アクチン生合成2019

    • Author(s)
      町田 幸大, 神澤 空流, 白子 太紀, 宮脇 翔馬, 今高 寛晃
    • Organizer
      大阪大学蛋白質研究所セミナー:化学によるタンパク質修飾の機能解析を目指して
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 再構成型タンパク質合成・フォールディング共役システムを利用したin vitroアクチン生合成2019

    • Author(s)
      町田幸大、白子太紀、井寄真仁、今高寛晃
    • Organizer
      第92回 日本生化学会年会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 再構成型タンパク質合成・フォールディング共役システムを利用した in vitroアクチン生合成2019

    • Author(s)
      町田 幸大, 宮脇 翔馬, 白子 太紀, 今高 寛晃
    • Organizer
      第14回 無細胞生命科学研究会
    • Related Report
      2019 Annual Research Report
  • [Presentation] アクチン遺伝病の試験管内解析2019

    • Author(s)
      町田幸大、神澤空流、白子太紀、今高寛晃
    • Organizer
      第42回 日本分子生物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] アクチン生合成の試験管内再現2019

    • Author(s)
      町田幸大、神澤空流、井寄真仁、白子太紀、今高寛晃
    • Organizer
      第4回 デザイン生命工学研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] アクチン遺伝病の試験管内解析2018

    • Author(s)
      町田幸大、神澤空流、白子太紀、今高寛晃
    • Organizer
      第41回 日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] アクチン遺伝病の試験管内解析2018

    • Author(s)
      町田幸大、神澤空流、白子太紀、今高寛晃
    • Organizer
      第13回無細胞生命科学研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] アクチン遺伝病の試験管内解析2018

    • Author(s)
      町田幸大、神澤空流、白子太紀、今高寛晃
    • Organizer
      細胞を創る研究会 11.0
    • Related Report
      2018 Annual Research Report
  • [Presentation] アクチン遺伝病の試験管内解析2018

    • Author(s)
      町田幸大、神澤空流、白子太紀、今高寛晃
    • Organizer
      第91回日本生化学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] In vitro analysis of actin fibrillization associated with developmental disorders2018

    • Author(s)
      Kodai Machida, Kuru Kanzawa, Taiki Hakushi, Hiroaki Imataka
    • Organizer
      International symposium on proteins; from the cradle to the grave
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 再構成型細胞様GUVを利用したポリグルタミンタンパク質の凝集と凝集抑制の解析2017

    • Author(s)
      町田 幸大, 神澤空流, 山本悠貴, 井寄真仁, 岡留壮志, 今高 寛晃
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] 再構成型ヒト細胞を利用したハンチンチン凝集解析システムの開発2017

    • Author(s)
      町田 幸大, 神澤空流, 山本悠貴, 井寄真仁, 今高 寛晃
    • Organizer
      第12回無細胞生命科学研究会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 再構成型人工細胞を利用したポリグルタミンタンパク質凝集の解析2017

    • Author(s)
      町田幸大
    • Organizer
      「新生鎖の生物学」平成29年度第1回班会議
    • Related Report
      2017 Annual Research Report
  • [Presentation] 人工細胞を用いた神経変性疾患の解析2017

    • Author(s)
      町田幸大、神澤空流、山本悠貴、井寄真仁、岡留壮志、今高寛晃
    • Organizer
      「細胞を創る」研究会 10.0
    • Related Report
      2017 Annual Research Report
  • [Presentation] PolyQ aggregation in GUV2017

    • Author(s)
      町田 幸大, 神澤空流, 今高 寛晃
    • Organizer
      「新生鎖の生物学」第4回 若手ワークショップ
    • Related Report
      2017 Annual Research Report
  • [Presentation] Polyglutamine-dependent protein aggregation in artificial cells2017

    • Author(s)
      Kodai Machida, Kuru Kanzawa, Yuki Yamamoto, Hiroaki Imataka
    • Organizer
      International Symposium on Protein Quality Control
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research

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Published: 2017-04-28   Modified: 2022-01-27  

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