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Biochemical study of the dicer-related helicases in C. elegans for anti-nematode drug discovery

Research Project

Project/Area Number 16K08229
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biological pharmacy
Research InstitutionToyohashi University of Technology

Principal Investigator

Eki Toshihiko  豊橋技術科学大学, 工学(系)研究科(研究院), 教授 (40192512)

Research Collaborator HIROSE yuu  
KOBAYASHI taishi  
MURAKAMI takuro  
KIMURA kosei  
SHIRAKI hayato  
SUGINO misato  
IIDA seiya  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2016: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords線虫 / ダイサー関連ヘリカーゼ / タンパク質間相互作用 / 抗線虫薬 / 核酸依存性ATPase / RNA干渉 / 創薬
Outline of Final Research Achievements

This study has aimed to clarify the biochemical properties of two RNAi factors DRH-3 (Dicer Related Helicase-3) and its paralog DRH-1 by purifying two DRH proteins. The fluorescence-based ATPase assay system was prepared and the double-stranded RNA-dependent ATPase activity of the purified DRH-3 was successfully found in this assay. I used this assay system to try the screening for potential DRH-3 inhibitors.
I prepared 14 GST-tagged mutated E1 proteins, four RNAi-related proteins and His- or His-SUMO-tagged DRH-3 to examine the protein-protein interaction between E1 (a novel protein with two Tudor domains) and DRH-3 by the pull-down method. The results suggested the Tudor domain/methylated residue-independent interaction between E1 and DRH-3 as well as the potential role of the N-terminal region of DRH-3 in the binding to E1.

Academic Significance and Societal Importance of the Research Achievements

本研究成果として、線虫の繁殖に必須なDRH-3が二本鎖RNA依存性ATPase活性を持つことを明らかにした。未だ生化学的解析のなされていないDRH-1の精製法を確立したことや、DRH-3とE1との相互作用解析からTudorドメインやDRH-3のN末領域に関して新規な知見が得られたことは、線虫DRHの機能を分子レベルで解明するための重要な手がかりとなり、RNA干渉研究における学術的意義がある。本研究を基にDRH-3のATPase活性阻害剤を探索することで、神経系を標的とする既存薬とは作用機序の異なる新規抗線虫薬を開発できる可能性があり、今後、社会的意義のある成果に結びつくことが期待される。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (9 results)

All 2019 2018 2017 2016

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (7 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Yeast-based genotoxicity tests for assessing DNA alterations and DNA stress responses: a 40-year overview2018

    • Author(s)
      Toshihiko Eki
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 102 Issue: 6 Pages: 2493-2507

    • DOI

      10.1007/s00253-018-8783-1

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Development and evaluation of yeast-based GFP and luciferase reporter assays for chemical-induced genotoxicity and oxidative damage2017

    • Author(s)
      Hajime Suzuki, Takahiro Sakabe, Yuu Hirose, and Toshihiko Eki
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 101 Issue: 2 Pages: 659-671

    • DOI

      10.1007/s00253-016-7911-z

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Presentation] 線虫ダイサー関連ヘリカーゼDRHの生化学的機能解析2019

    • Author(s)
      村上匠蔵,小林大士,木村康誠,広瀬 侑,浴 俊彦
    • Organizer
      第83回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] 線虫ダイサー関連ヘリカーゼDRH-1およびDRH-3の生化学的解析2018

    • Author(s)
      小林大士,木村康誠,白木 隼,広瀬 侑, 浴 俊彦
    • Organizer
      第82回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2018 Annual Research Report 2017 Research-status Report
  • [Presentation] 線虫のダイサー関連ヘリカーゼDRH-1とDRH-3の生化学的研究2018

    • Author(s)
      小林大士、村上匠蔵、木村康誠、白木 隼、広瀬 侑、浴 俊彦
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 2つの線虫ダイサー関連ヘリカーゼDRHの発現・精製と生化学的解析2018

    • Author(s)
      浴 俊彦、小林大士、木村康誠、白木 隼、杉野美里、広瀬 侑
    • Organizer
      日本薬学会第138年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 線虫ダイサー関連ヘリカーゼDRH-1およびDRH-3の生化学的解析2017

    • Author(s)
      浴 俊彦、小林大士、木村康誠、白木 隼、杉野美里、飯田誠也、広瀬 侑
    • Organizer
      2017年度生命科学系学会合同年次大会(ConBio2017)
    • Related Report
      2017 Research-status Report
  • [Presentation] Development of the yeast-based reporter assay for sensing genotoxicity and oxidative toxicity and improvement of yeast strains by genome editing2016

    • Author(s)
      Toshihiko Eki, Hajime Suzuki, Shogo Matsunobu, Yuki Miura, Takahiro Sakabe, and Yuu Hirose
    • Organizer
      The Irago conference 2016
    • Place of Presentation
      Chofu (Tokyo, Japan)
    • Year and Date
      2016-11-01
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Development and characterization of the reporter gene assays in yeast for detecting oxidative stress and DNA damage2016

    • Author(s)
      Toshihiko Eki, Hajime Suzuki, Takahiro Sakabe, Yuu Hirose
    • Organizer
      6th International Conference on Plasma Medicine (ICPM-6)
    • Place of Presentation
      Bratislava, Slovakia
    • Year and Date
      2016-09-04
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research

URL: 

Published: 2016-04-21   Modified: 2020-03-30  

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