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Direct single-molecule observations of DNA unwinding by SV40 large tumor antigen under a negative DNA supercoil state

Research Project

Project/Area Number 15K06946
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Molecular biology
Research InstitutionGunma University

Principal Investigator

Oshige Masahiko  群馬大学, 大学院理工学府, 准教授 (00451716)

Project Period (FY) 2015-10-21 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
KeywordsDNA / DNA複製 / 高次構造 / 超らせん / SV40 / 1分子観察 / 1分子解析 / 複製起点 / 複製開始 / DNAポリメラーゼ / DNA合成反応 / T4 DNAリガーゼ / 1分子計測 / 分子計測 / 分子操作 / 可視化技術 / 酵素反応
Outline of Final Research Achievements

Superhelices, which are induced by the twisting and coiling of double-helical DNA in chromosomes, are thought to affect DNA metabolic processes. In this study, I report the effects of negative supercoiling on the unwinding activity of simian virus 40 large tumor antigen (SV40 TAg) at a single-molecular level. The supercoiling density of linear DNA templates was controlled using magnetic tweezers and monitored using a fluorescent microscope in a flow cell. SV40 TAg-mediated DNA unwinding under relaxed and negative supercoil states was analyzed by the direct observation of both single- and double-stranded regions of single DNA molecules. Increased negative superhelicity stimulated SV40 TAg-mediated DNA unwinding more strongly than a relaxed state; furthermore, negative superhelicity was associated with an increased probability of SV40 TAg-mediated DNA unwinding. These results suggest that negative superhelicity helps to regulate the initiation of DNA replication.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (10 results)

All 2017 2016 2015

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (9 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Journal Article] Direct single-molecule observations of DNA unwinding by SV40 large tumor antigen under a negative DNA superhelical state.2017

    • Author(s)
      Takahashi S, Motooka S, Kawasaki S, Kurita H, Mizuno T, Matsuura SI, Hanaoka F, Mizuno A, Oshige M, Katsura S
    • Journal Title

      J Biomol Struct Dyn

      Volume: 5 Issue: 1 Pages: 1-13

    • DOI

      10.1080/07391102.2016.1269689

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 生体分子を操作する2017

    • Author(s)
      大重真彦
    • Organizer
      静電気学会2017年度シンポジウム
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] SSB-YFPを用いたDNAポリメラ-ゼの1分子解析2017

    • Author(s)
      勝見匡、栗本雅之、高橋俊介、川崎祥平、栗田弘史、松浦俊一、水野彰、大重真彦、桂進司
    • Organizer
      2017年度生命科学系学会合同年次大会(ConBio2017)
    • Related Report
      2017 Annual Research Report
  • [Presentation] 1分子レベルの蛍光観察による負の超らせんがSV40DNA複製反応の開始に与える影響の評価2016

    • Author(s)
      高橋俊介、本岡伸也、川崎祥平、栗田弘史、水野武、松浦俊一、花岡文雄、水野彰、大重真彦、桂進司
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-12-02
    • Related Report
      2016 Research-status Report
  • [Presentation] ヒトDNAポリメラーゼδ触媒サブユニットの1分子観察の試み2016

    • Author(s)
      石川裕一、高橋俊介、岸川健太、川崎祥平、栗田弘史、水野武、松浦俊一、水野彰、大重真彦、桂進司
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-12-02
    • Related Report
      2016 Research-status Report
  • [Presentation] 蛍光T4 DNA Ligaseの1分子解析の試み2016

    • Author(s)
      小和瀬聡実、碓井智大、高橋俊介、川崎祥平、栗田弘史、水野彰、大重真彦、桂進司
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-12-02
    • Related Report
      2016 Research-status Report
  • [Presentation] 大腸菌由来DNAポリメラ-ゼによるDNA合成反応の1分子解析2016

    • Author(s)
      柳基成、高橋俊介、川崎祥平、栗田弘史、松浦俊一、水野彰、大重真彦、桂進司
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-12-02
    • Related Report
      2016 Research-status Report
  • [Presentation] DNAの負の超らせん導入下におけるSV40ラージT抗原によるDNA解鎖反応の1分子直接観察2015

    • Author(s)
      高橋俊介、本岡伸也、川崎祥平、栗田弘史、水野武、松浦俊一、水野彰、大重真彦、桂進司
    • Organizer
      BMB2015(第38回分子生物学会年会・第88回生化学会大会合同大会)
    • Place of Presentation
      神戸ポートアイランド(兵庫県・神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] 蛍光複製タンパク質を用いた1本鎖DNA標識によるDNA合成反応の1分子観察2015

    • Author(s)
      高橋俊介、石川裕一、柳基成、小和瀬聡実、川崎祥平、栗田弘史、水野武、松浦俊一、水野彰、大重真彦、桂 進司
    • Organizer
      BMB2015(第38回分子生物学会年会・第88回生化学会大会合同大会)
    • Place of Presentation
      神戸ポートアイランド(兵庫県・神戸市)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] Direct single-molecule observations of DNA synthesis and digestion reactions.2015

    • Author(s)
      Shunsuke Takahashi, Shohei Kawasaki, Hirofumi Kurita, Takeshi Mizuno, Shun-ichi Matsuura, Akira Mizuno, Masahiko Oshige, Shinji Katsura
    • Organizer
      The 11th International Workshop on Radiation Effects on Semiconductor Devices for Space Applications (11th RASEDA) and The 7th International Conference on Advanced Micro-Device Engineering (7th AMDE)
    • Place of Presentation
      桐生市市民文化会館(群馬県・桐生市)
    • Year and Date
      2015-11-12
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research

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Published: 2015-10-21   Modified: 2019-03-29  

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