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Development of novel labeling technique for macro molecule structure analysis

Research Project

Project/Area Number 24651151
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Nanomaterials/Nanobioscience
Research InstitutionKyushu University

Principal Investigator

MAYANAGI Kouta  九州大学, 生体防御医学研究所, 助教 (50418571)

Co-Investigator(Renkei-kenkyūsha) ISHINO Yoshizumi  九州大学, (連合)農学研究科(研究院), 教授 (30346837)
Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywords電子顕微鏡 / ナノバイオ / 可視化 / 生物物理 / 蛋白質 / DNA / 分子標識 / 画像解析
Outline of Final Research Achievements

Molecular labeling is a key technique for mapping individual components in the 3D map of supermolecules, obtained by single particle analysis. We developed novel methods of labeling DNA in the DNA-protein complexes, which are biologically very important. We used streptavidin in our previous work, which was too large (about 60k for tetramer) for more detailed analysis.
In this project we tried mainly two methods. The first uses the nano structures of DNA for labeling. Using the pinpoint shape of the nano-structure, we could develop a label, which could point the DNA terminal more precisely.
Another label is a small bulky structure, the size of which is comparable to the resolution of the single particle analysis. As the second method seemed to be more promising, we applied the later method to the protein-DNA complexes labeling experiment, and could successfully visualize the label in the total map of the complex.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (12 results)

All 2014 2013 2012

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (11 results) (of which Invited: 3 results)

  • [Journal Article] The centromeric nucleosome-like CENP-T-W-S-X complex induces positive supercoils into DNA.2013

    • Author(s)
      Kozo Takeuchi, Tatsuya Nishino, Kouta Mayanagi, Naoki Horikoshi, Akihisa Osakabe, Hiroaki Tachiwana, Tetsuya Hori, Hitoshi Kurumizaka and Tatsuo Fukagawa.
    • Journal Title

      Nucleic Acids Research

      Volume: 42 Issue: 3 Pages: 1644-1655

    • DOI

      10.1093/nar/gkt1124

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] DNA複製フォーク複合体の分子構築及び切換え機構の研究2014

    • Author(s)
      真柳浩太
    • Organizer
      タンパク質研究所セミナー
    • Place of Presentation
      大阪
    • Year and Date
      2014-02-07 – 2014-02-08
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Divergently evolved activation mechanism of the replicative helicase in the thermophilic archaeon, Thermoplasma acidophilum2013

    • Author(s)
      Hiromi Ogino, Sonoko Ishino, Kouta Mayanagi, Takuji Oyama, Tsuyoshi Shirai, Kosuke Morikawa, Gyri Teien Haugland, Nils-Kare Birkeland, Daisuke Kohda, Yoshizumi Ishino
    • Organizer
      第36回日本分子生物学会
    • Place of Presentation
      神戸
    • Year and Date
      2013-12-03 – 2013-12-06
    • Related Report
      2013 Annual Research Report
  • [Presentation] DNA 複製に関与する超分子複合体の単粒子解析2013

    • Author(s)
      真柳浩太
    • Organizer
      日本顕微鏡学会第57回シンポジウム
    • Place of Presentation
      名古屋
    • Year and Date
      2013-11-15 – 2013-11-16
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 好熱性アーキアThermoplasma acidophilumにおける複製ヘリカーゼ活性化機構の分岐進化2013

    • Author(s)
      尾木野 弘実, 石野 園子, 真柳 浩太, 大山 拓次, 白井 剛, 森川 耿右, Haugland Gyri Teien, Birkeland Nils-Kare, 神田 大輔, 石野 良純
    • Organizer
      第14回極限環境生物学会
    • Place of Presentation
      川崎
    • Year and Date
      2013-10-26 – 2013-10-27
    • Related Report
      2013 Annual Research Report
  • [Presentation] Electron microscopic structure analysis of protein-DNA complexes2013

    • Author(s)
      Kouta Mayanagi
    • Organizer
      The 3rd International conference on New Frontier of the Research on RecA-family recombinases and their accessory proteins
    • Place of Presentation
      Nantes, France
    • Year and Date
      2013-10-03 – 2013-10-05
    • Related Report
      2013 Annual Research Report
  • [Presentation] 組み替えhuman poly(ADP-ribose) polymerase 1の精製と予備的構造解析2013

    • Author(s)
      Kenichi Koyama, Kouta Mayanagi, Takayuki Eguchi, Hiroyuki Morita, Kazuo Kamemura, Yoshisuke Nishi, Masanao Miwa Tsuyoshi Shirai
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Year and Date
      2013-09-11 – 2013-09-13
    • Related Report
      2013 Annual Research Report
  • [Presentation] Activation mechanism of the replicative helicase in the thermophilic archaeon, Thermoplasma acidophilum2013

    • Author(s)
      Hiromi Ogino, Sonoko Ishino, Kouta Mayanagi, Takuji Oyama, Tsuyoshi Shirai, Kosuke Morikawa, Gyri Teien Haugland, Nils-Kare Birkeland, Daisuke Kohda, Yoshizumi Ishino
    • Organizer
      Thermophiles 2013
    • Place of Presentation
      Regensburg, Germany
    • Year and Date
      2013-09-08 – 2013-09-13
    • Related Report
      2013 Annual Research Report
  • [Presentation] Electron microscopic analysis of molecular architecture and switching mechanism of DNA replication fork complex2013

    • Author(s)
      Kouta Mayanagi, Shinichi Kiyonari, Hirokazu Nishida, Sonoko Ishino, Mihoko Saito, Daisuke Kohda, Yoshizumi Ishino, Tsuyoshi Shirai, Kosuke Morikawa
    • Organizer
      ICSG2013-SLS
    • Place of Presentation
      札幌
    • Year and Date
      2013-07-30 – 2013-08-01
    • Related Report
      2013 Annual Research Report
  • [Presentation] Molecular architecture and switching mechanism of DNA replication fork complex2013

    • Author(s)
      Kouta Mayanagi
    • Organizer
      Gordon Research Conference,Three Dimensional Electron Microscopy
    • Place of Presentation
      Colby-Sawyer College, NH, USA
    • Year and Date
      2013-06-23 – 2013-06-28
    • Related Report
      2013 Annual Research Report
  • [Presentation] 単粒子解析による複製関連複合体の制御機構の研究2012

    • Author(s)
      真柳浩太
    • Organizer
      遺伝研研究集会
    • Place of Presentation
      遺伝学研究所
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] Thermoplasma acidophilum由来Cdc6/Orc1とホモ4量体のGINSによるMCMとの相乗的な活性化2012

    • Author(s)
      尾木野弘実、石野園子、真柳浩太、大山拓次、白井剛、森川耿右、Gyri Teien Haugland、Nils-Kare Birkeland、石野良純
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場、マリンメッセ福岡
    • Related Report
      2012 Research-status Report

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Published: 2013-05-31   Modified: 2019-07-29  

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