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Study of regulation of DNA replication by controlling superhelicities of single DNA molecules

Research Project

Project/Area Number 25650002
Research Category

Grant-in-Aid for Challenging Exploratory Research

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

Principal Investigator

Shinji Katsura  群馬大学, 大学院理工学府, 教授 (10260598)

Co-Investigator(Kenkyū-buntansha) Oshige Masahiko  群馬大学, 大学院理工学府, 准教授 (00451716)
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords1分子観察 / DNA2次構造 / 超らせん / マグネチックツィーザー / 磁気ピンセット / 微細流路 / 1本鎖DNA / DNA複製制御 / DNA2次構造 / DNA複製 / DNA形態制御 / DNA超らせん
Outline of Final Research Achievements

We developed an apparatus based on magnetic tweezers to introduce supercoil on a single DNA molecule, and studied the effects of negative superhelicities on the local DNA denaturation in λDNA containing the SV40 origin of DNA replication. Furthermore, we also studied the effects of superhelicities on the local denaturation in the presence of SV40 Large T antigen(SV40TAg), which is an initiation protein of SV40 DNA replication,
Even in the absence of SV40TAg, the local denaturation of SV40ori-λDNA under a negative supercoil state was observed around the replication origin and the probability of occurrence of the local denaturation increased with negative superhelicity. Furthermore, addition of SV40TAg enhanced the probability of occurrence of the local denaturation and extended the single-stranded region under higher negative superhelicities.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (13 results)

All 2015 2014 2013

All Journal Article (4 results) (of which Peer Reviewed: 4 results,  Acknowledgement Compliant: 1 results) Presentation (9 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Direct Single-Molecule Observations of Local Denaturation of a DNA Double Helix under a Negative Supercoil State.2015

    • Author(s)
      Takahashi S, Motooka S, Usui T, Kawasaki S, Miyata H, Kurita H, Mizuno T, Matsuura S, Mizuno A, Oshige M, Katsura S.
    • Journal Title

      Analytical Chemistry

      Volume: 87 Issue: 6 Pages: 3490-3497

    • DOI

      10.1021/acs.analchem.5b00044

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Real-time single-molecule observations of T7 Exonuclease activity in a microflow channel2014

    • Author(s)
      S. Takahashi, T. Usui, S. Kawasaki, H. Miyata, H. Kurita, S.-i. Matsuura, A. Mizuno, M. Oshige, and S. Katsura
    • Journal Title

      Analytical Biochemistry

      Volume: - Pages: 24-30

    • DOI

      10.1016/j.ab.2014.04.012

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] A New Direct Single-Molecule Observation Method for DNA Synthesis Reaction Using Fluorescent Replication Protein A2014

    • Author(s)
      S. Takahashi, S. Kawasaki, H. Miyata, H. Kurita, T. Mizuno, S.-i. Matsuura, A. Mizuno, M. Oshige, and S. Katsura
    • Journal Title

      Sensors

      Volume: 14 Issue: 3 Pages: 5174-5182

    • DOI

      10.3390/s140305174

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Direct observation of fluorescently labeled single-stranded λDNA molecules in a micro-flow channel2013

    • Author(s)
      S. Takahashi, S. Kawasaki, K. Yamaguchi, H. Miyata, H. Kurita, T. Mizuno, S. Matsuura, A. Mizuno, M. Oshige, and S. Katsura
    • Journal Title

      Journal of Fluorescence

      Volume: - Issue: 4 Pages: 635-640

    • DOI

      10.1007/s10895-013-1210-1

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [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回日本分子生物学会年会・第88回日本生化学会大会 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 Annual Research Report
  • [Presentation] DNAおよびDNA代謝酵素の1分子蛍光観察による動態解析2015

    • Author(s)
      高橋俊介、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
      化学工学会群馬大会2015
    • Place of Presentation
      桐生市民文化会館(群馬県桐生市)
    • Year and Date
      2015-11-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] Direct Single-Molecule Observations of DNA Synthesis and Digestion Reactions2015

    • Author(s)
      Takahashi, S., Kawasaki, S., Kurita, H., Mizuno, T., Matsuura, S-I., Mizuno, A., Oshige, M., Katsura, S.
    • Organizer
      The 11th International Workshop on Radiation Effects on Semiconductor Devices for Space Applications & The 7th International Conference on Advanced Micro-Device Engineering
    • Place of Presentation
      City Performing Arts Center (Kiryu, Gunma, Japan)
    • Year and Date
      2015-11-11
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 負の超らせん状態でのSV40ラージT抗原によるDNA解鎖の1分子観察2015

    • Author(s)
      高橋俊介、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23回DNA複製・組換え・修復ワークショップ
    • Place of Presentation
      焼津グランドホテル(静岡県焼津市)
    • Year and Date
      2015-10-19
    • Related Report
      2015 Annual Research Report
  • [Presentation] 負の超らせんDNAによる二重らせんの局所的な開裂の1分子観察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
      2015年度生物工学若手研究者の集い(若手会)夏のセミナー
    • Place of Presentation
      タナベ名古屋研修センター(愛知県北名古屋市)
    • Year and Date
      2015-07-11
    • Related Report
      2015 Annual Research Report
  • [Presentation] Real-time single-molecule observations of T7 Exonuclease activity in a microflow channel,2014

    • Author(s)
      S. Takahashi, T. Usui, S. Kawasaki, H. Miyata, H. Kurita, S. Matsuura, A. Mizuno, M. Oshige, S. Katsura
    • Organizer
      1st International Symposium of Gunma University Medical Innovation and 6th International Conference on Advanced Micro-Device Engineering (GUMI&AMDE2014)
    • Place of Presentation
      Kiryu, Gunma, JAPAN
    • Year and Date
      2014-12-05
    • Related Report
      2014 Research-status Report
  • [Presentation] 負の超らせん導入による二重らせんの局所的な開裂の1分子直接観測2014

    • Author(s)
      高橋俊介, 本岡伸也, 川崎祥平, 宮田英史, 栗田弘史, 松浦俊一, 水野武, 水野彰, 大重真彦, 桂進司
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      横浜、神奈川
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Direct observation of fluorescently labeled single-stranded λDNA molecules in a micro-flow channel2013

    • Author(s)
      S. Takahashi, S. Kawasaki, K. Yamaguchi, H. Miyata, H. Kurita, T. Mizuno, S. Matsuura, A. Mizuno, M. Oshige and S, Katsura
    • Organizer
      5th International Conference on Advanced Micro-Device Engineering
    • Place of Presentation
      Kiryu, Gunma, Japan
    • Related Report
      2013 Research-status Report
  • [Presentation] 蛍光複製タンパク質A を用いた1本鎖DNA 標識によるDNA 合成反応のリアルタイム1分子観察2013

    • Author(s)
      高橋 俊介,川崎 祥平,宮田 英史,栗田 弘史,水野 武,松浦 俊一,水野 彰,大重 真彦,桂 進司
    • Organizer
      第36回日本分子生物学会
    • Place of Presentation
      神戸ポートアイランド
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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