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Super-resolved single-molecule imaging of nuclear proteins for quantification of phase separated behavior

Research Project

Project/Area Number 18K14661
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 43040:Biophysics-related
Research InstitutionTokyo Institute of Technology

Principal Investigator

Ito Yuma  東京工業大学, 生命理工学院, 助教 (70803245)

Project Period (FY) 2018-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords1分子計測 / 生細胞イメージング / 相分離 / 細胞核 / 1分子計測 / 生細胞1分子イメージング / 液相分離
Outline of Final Research Achievements

We established a method for high-resolution single-molecule imaging in living cell. This method enables us to quantify directly the phase separated behavior of nuclear proteins. We demonstrated the analysis of ribosome biogenesis dependent behavior of nucleolar proteins and chromocenter independent behavior of heterochromatin proteins. These results suggest that this method provides new quantitative insights into viscoelastic dynamics of nuclear proteins in the living cell that are difficult to achieve using previous approaches.

Academic Significance and Societal Importance of the Research Achievements

細胞内の相分離は様々な生物学的機能に関わる極めて重要な性質と考えられているが、その動態解析にはタンパク質の抽出精製等による試験管内での実験が一般的であった。本研究の高精細な1分子イメージング技術開発により、生きた細胞の小器官内で直接相分離挙動を定量解析することが可能になった。本研究の技術は、今まで未知であった生細胞内における生体分子の相分離動態の解明にむけて、強力な手法になると期待できる。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • Research Products

    (14 results)

All 2019 2018 Other

All Journal Article (3 results) (of which Peer Reviewed: 2 results,  Open Access: 1 results) Presentation (10 results) (of which Invited: 2 results) Remarks (1 results)

  • [Journal Article] 生細胞内の1分子動態と相互作用の定量解析2019

    • Author(s)
      伊藤由馬、徳永万喜洋
    • Journal Title

      パリティ

      Volume: 34 Pages: 72-74

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] 全反射照明蛍光顕微鏡 ― 対物レンズ型2019

    • Author(s)
      徳永万喜洋、伊藤由馬
    • Journal Title

      実験医学増刊

      Volume: 36 Pages: 36-37

    • Related Report
      2018 Research-status Report
  • [Journal Article] CLIP-170 is essential for MTOC repositioning during T cell activation by regulating dynein localisation on the cell surface2018

    • Author(s)
      Lim Wei Ming、Ito Yuma、Sakata-Sogawa Kumiko、Tokunaga Makio
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 17747-17747

    • DOI

      10.1038/s41598-018-35593-z

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 超解像1分子イメージングを用いたクロマチンタンパク質の動態と局在の定量解析2019

    • Author(s)
      伊藤 由馬、徳永 万喜洋
    • Organizer
      第19回日本蛋白質科学会年会・第71回日本細胞生物学大会 合同年次大会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Molecular localization and dynamics of Mediator regulating transcription elongation using single-molecule and super-resolution microscopy2019

    • Author(s)
      Yuma Ito, Shinnosuke Kunimi, Hidehisa Takahashi, Makio Tokunaga
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Single-molecule dynamics and localization of nucleolar proteins in phase-separated compartments of nucleolus2019

    • Author(s)
      Supanut Sirisukhodom, Daiki Matsumoto, Yuma Ito, Noriko Saitoh, Kumiko Sakata-Sogawa, Makio Tokunaga
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Dynamics and localization of Heterochromatin protein 1α involved in phase separation using single-molecule and super-resolution imaging2019

    • Author(s)
      Takahiro Maeda, Yuma Ito, Shin-Ya Isobe, Chikashi Obuse, Makio Tokunaga
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] A single-molecule localization approach to quantify the interaction between transcriptional machinery and chromatin structure2019

    • Author(s)
      Yuma Ito, Makio Tokunaga
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 蛍光1分子イメージングにおける動態解析法の開発と核内タンパク質挙動の定量解析2018

    • Author(s)
      伊藤由馬
    • Organizer
      日本生化学会 第91回大会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] CLIP-170 is essential for MTOC repositioning during T cell activation by recruiting proteins to the plus-end tracking on the cell surface2018

    • Author(s)
      Lim WM, Ito Y, Sakata-Sogawa K, Tokunaga M
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Dimerization dependent single-molecule dynamics of MafG transcription factor in living cell2018

    • Author(s)
      Ito Y, Maeda T, Sakata-Sogawa Kumiko, Shiina M, Tokunaga M
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Dynamics of Heterochromatin protein 1α in living cells using single-molecule imaging2018

    • Author(s)
      Maeda T, Ito Y, Isobe S, Obuse C, Tokunaga M
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Single-molecule imaging of post-translational modification using genetically encoded antibody probe2018

    • Author(s)
      Sato S, Ito Y, Sato Y, Kimura H, Tokunaga M
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Remarks] 東京工業大学 生命理工学院 徳永研究室

    • URL

      http://www.toku.bio.titech.ac.jp/

    • Related Report
      2019 Annual Research Report

URL: 

Published: 2018-04-23   Modified: 2021-02-19  

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