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標的DNA検索機構の理解に向けた核内環境のライブセルイメージング

Research Project

Project/Area Number 16J07205
Research Category

Grant-in-Aid for JSPS Fellows

Allocation TypeSingle-year Grants
Section国内
Research Field Molecular biology
Research InstitutionNational Institute of Genetics

Principal Investigator

野﨑 慎  国立遺伝学研究所, 遺伝メカニズム研究系, 特別研究員(PD) (40780185)

Project Period (FY) 2016-04-22 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsクロマチン / 転写因子 / ライブセルイメージング / 1分子イメージング / 減数分裂 / ヌクレオソーム / 核 / 超解像イメージング / 標的DNA検索
Outline of Annual Research Achievements

本研究の目的は,細胞内において標的DNAがどのように検索されるかという機構の理解を進めることである.2016年度と2017年度には,生細胞クロマチンの超解像イメージングと,転写因子1分子イメージングの開発を進め,クロマチンとタンパク質によるDNA情報検索に関する理解を深めてきた.それらの観察の結果,クロマチンの密な構造(クロマチンドメイン)が,核空間における転写因子による検索可能領域の限定に重要であることが示唆された.2018年度は,さらに踏み込んだ標的DNA検索の研究を行うため,ハーバード大学のNancy Klecknerラボに滞在し,減数分裂期における相同染色体検索の研究を行ってきた.①減数分裂期で,「いつ」「どのように」相同染色体が相互作用し,②「どのように」相同配列を検索するかについて明らかにすることを解くべき問題として設定した.この問題にアプローチするために,出芽酵母において,減数分裂期におけるプログラムDSB(DNA二本鎖切断)が比較的高い頻度で起きるホットスポット周辺のクロマチンを蛍光タンパク質によって染色し,その領域がお互いをどのように検索しているのかについて2色ライブセルイメージを行なった.様々な試行錯誤を繰り返した結果,減数分裂期中のDSBホットスポットの動きを追跡することが可能となった.その結果,相同染色体間の減数分裂前後における相互作用を観察することができた.

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Report

(3 results)
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • Research Products

    (12 results)

All 2019 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (6 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 5 results,  Open Access: 4 results,  Acknowledgement Compliant: 2 results) Presentation (5 results) (of which Int'l Joint Research: 4 results)

  • [Int'l Joint Research] ハーバード大学(米国)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Single nucleosome imaging reveals loose genome chromatin networks via active RNA polymerase II2019

    • Author(s)
      R Nagashima, K Hibino, SS Ashwin, M Babokhov, S Fujishiro, R Imai, T Nozaki, S Tamura, T Tani, H Kimura, M Shribak, MT Kanemaki, M Sasai, K Maeshima
    • Journal Title

      Journal of Cell Biology

      Volume: 218 Issue: 5 Pages: 1511-1530

    • DOI

      10.1083/jcb.201811090

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Dynamic chromatin folding in the cell2018

    • Author(s)
      Tadasu Nozaki, Damien F. Hudson, Sachiko Tamura, Kazuhiro Maeshima
    • Journal Title

      Nuclear Architecture and Dynamics

      Volume: 1 Pages: 101-122

    • DOI

      10.1016/b978-0-12-803480-4.00004-1

    • ISBN
      9780128034804
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Journal Article] Dynamic organization of chromatin domains revealed by super-resolution live-cell imaging.2017

    • Author(s)
      Nozaki, T., Imai, R., Tanbo, M., Nagashima, R., Tamura, S., Tani, T., Joti, Y., Tomita, M., Hibino, K., Wendt, K.S., Okada, Y., Nagai, T., Maeshima, K.
    • Journal Title

      Molecular Cell.

      Volume: 67 Issue: 2 Pages: 282-293

    • DOI

      10.1016/j.molcel.2017.06.018

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Density imaging of heterochromatin in live cells using orientation-independent-DIC microscopy.2017

    • Author(s)
      Imai, R., Nozaki, T., Tani, T., Kaizu, K., Hibino, K., Ide, S., Tamura, S., Takahashi, K., Shribak, M., and Maeshima, K.
    • Journal Title

      Molecular Biology of the Cell.

      Volume: 28 Issue: 23 Pages: 3349-3359

    • DOI

      10.1091/mbc.e17-06-0359

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Bridging the dynamics and organization of chromatin domains by mathematical modeling2017

    • Author(s)
      Soya Shinkai, Tadasu Nozaki, Kazuhiro Maeshima, Yuichi Togashi
    • Journal Title

      Nucleus

      Volume: 印刷中

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Dynamic Nucleosome Movement Provides Structural Information of Topological Chromatin Domains in Living Human Cells2016

    • Author(s)
      Shinkai S, Nozaki T, Maeshima K, Togashi Y
    • Journal Title

      PLoS Comput Biol

      Volume: 12 Issue: 10 Pages: e1005136-e1005136

    • DOI

      10.1371/journal.pcbi.1005136

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 4D live-cell imaging of DSB independent pairing between homologous chromosomes in budding yeast2018

    • Author(s)
      Tadasu Nozaki and Nancy Kleckner
    • Organizer
      Cold Spring Harbor meeting: Single Biomolecules
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Live-cell imaging of pre-meiotic pairing between homologous chromosomes2018

    • Author(s)
      Tadasu Nozaki and Nancy Kleckner
    • Organizer
      Meiosis Conference GRC
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Dynamic organization of chromatin domains revealed by super resolution live-cell imaging2017

    • Author(s)
      Tadasu Nozak, Kazuhiro Maeshima
    • Organizer
      EMBO Conference on Meiosis 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Dynamic organization of chromatin domains revealed by super-resolution imaging2016

    • Author(s)
      Tadasu Nozaki and Kazuhiro Maeshima
    • Organizer
      2016 Workshop on DNA Reactions and DNA/Chromosome Dynamics
    • Place of Presentation
      Woods Hole (USA)
    • Year and Date
      2016-09-11
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Dynamic chromatin domains revealed by super-resolution live-cell imaging2016

    • Author(s)
      野崎慎, 前島一博
    • Organizer
      第54回日本生物物理学会年会
    • Place of Presentation
      筑波
    • Related Report
      2016 Annual Research Report

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Published: 2016-05-17   Modified: 2024-03-26  

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