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Elucidation of chromosome structure by DNA multi-color imaging using super-resolution microscopy in living cells

Research Project

Project/Area Number 16H06212
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Applied molecular and cellular biology
Research InstitutionNational Institute of Advanced Industrial Science and Technology

Principal Investigator

Hideaki Takata  国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (20455207)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥16,770,000 (Direct Cost: ¥12,900,000、Indirect Cost: ¥3,870,000)
Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2016: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Keywords染色体 / クロマチン / イメージング / カルシウム / リン酸化 / CRISPR imaging / ゲノム編集 / CRISPR/Cas9 / 細胞分裂 / 生細胞イメージング / DNA相互作用
Outline of Final Research Achievements

To elucidate the mechanism of chromosome condensation during mitosis, chromatin imaging techniques including FLIM-FRET, super-resolution microscopy, and CRISPR/dCas9 system were employed. As a result, the function of a divalent cation, calcium ion, was revealed as a booster of chromosome condensation after nuclear envelope breakdown. In addition to the calcium ion, KIF4A which is localized at chromosome scaffold, an axial structure observed in the center of sister chromatids, is required for chromosome condensation during early mitosis. The phosphorylation site of KIF4A by Cdk1 was identified, and the phosphorylation was found to be required for the interaction with condensin I complex, which is also localized at chromosome scaffold and induces chromosome condensation by chromatin loop formation. The changes in chromatin structure in cancer cells and senescence cells were also detected using a CRISPR/dCas9 system.

Academic Significance and Societal Importance of the Research Achievements

本研究により、分裂期における染色体凝縮には蛋白質だけでなくカルシウムイオンの働きも必要であることを示した。このため、細胞内カルシウムイオン制御に関わる遺伝子異常が染色体異常につながる可能性が考えられる。また、KIF4AのCdk1によるリン酸化が染色体凝縮を開始するカギの一つであることを示し、分裂期初期の染色体凝縮機構の解明が進んだ。さらに、本研究で構築したCRISP/dCas9システムを活用することで、細胞状態変化に伴うクロマチン構造変化と遺伝子発現変化との関連を調べることで、将来的に生命現象の理解や疾患モニタリングへの応用等が期待できる。

Report

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

    (14 results)

All 2018 2017 2016 Other

All Journal Article (4 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 4 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Presentation (8 results) (of which Int'l Joint Research: 4 results,  Invited: 4 results) Remarks (2 results)

  • [Journal Article] Cdk1-dependent phosphorylation of KIF4A at S1186 triggers lateral chromosome compaction during early mitosis2018

    • Author(s)
      Hideaki Takata, Marliza Madung, Kaoru Katoh, Kiichi Fukui
    • Journal Title

      PLoS ONE

      Volume: 13(12) Issue: 12 Pages: e0209614-e0209614

    • DOI

      10.1371/journal.pone.0209614

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Calcium depletion destabilises kinetochore fibres by the removal of CENP-F from the kinetochore2017

    • Author(s)
      Phengchat Rinyaporn、Takata Hideaki、Uchiyama Susumu、Fukui Kiichi
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 7355-7355

    • DOI

      10.1038/s41598-017-07777-6

    • NAID

      120006995023

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Interdependency and phosphorylation of KIF4 and condensin I are essential for organization of chromosome scaffold2017

    • Author(s)
      Poonperm Rawin、Takata Hideaki、Uchiyama Susumu、Fukui Kiichi
    • Journal Title

      PLOS ONE

      Volume: 12 Issue: 8 Pages: e0183298-e0183298

    • DOI

      10.1371/journal.pone.0183298

    • NAID

      120006995026

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Calcium ions function as a booster of chromosome condensation.2016

    • Author(s)
      Phengchat, R., Takata, H., Morii, K., Inada, N., Murakoshi, H., Uchiyama, S., and Fukui, K.
    • Journal Title

      Scientific Reports

      Volume: 6 Issue: 1 Pages: 38281-38281

    • DOI

      10.1038/srep38281

    • NAID

      120006995007

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Presentation] Chromosome structural analysis using imaging techniques2018

    • Author(s)
      Hideaki Takata
    • Organizer
      13th India-Japan Bilateral Conference
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Chromosomal proteins required for authentic chromosome morphology.2018

    • Author(s)
      Hideaki Takata
    • Organizer
      2nd e-ASIA Symposium on Functional Nano-Biology
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 超解像イメージングにより明らかとなった新たな染色体構造2018

    • Author(s)
      高田英昭, Rawin Poonperm, 加藤薫, 松田厚志, 平岡泰, 福井希一
    • Organizer
      第27回日本バイオイメージング学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Studies on the condensation mechanism of mitotic chromosomes and its biological significance2018

    • Author(s)
      Hideaki Takata
    • Organizer
      6th Asia-Pacific Chromosome Colloquium
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Imaging of chromatin condensation by FLIM-FRET reveals new calcium functions during mitosis2018

    • Author(s)
      Hideaki Takata
    • Organizer
      Workshop in The First RCB Bioimaging School
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 高感度DNA凝縮評価システムにより明らかとなったカルシウムイオンの新たな機能2018

    • Author(s)
      Hideaki Takata
    • Organizer
      関西バイオ医療研究会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 分裂期染色体におけるカルシウムイオンの機能解析2017

    • Author(s)
      Hideaki Takata
    • Organizer
      染色体学会第68回年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] FLIM-FRETを用いた染色体凝縮における二価陽イオンの機能解析2017

    • Author(s)
      Hideaki Takata
    • Organizer
      第69回日本細胞生物学会大会
    • Related Report
      2017 Annual Research Report
  • [Remarks] カルシウムイオンの欠乏が染色体異常を引き起こす原因を解明

    • URL

      http://www.aist.go.jp/aist_j/press_release/pr2017/pr20170807/pr20170807.html

    • Related Report
      2017 Annual Research Report
  • [Remarks] カルシウムイオンの欠乏が細胞分裂異常を誘導する

    • URL

      http://resou.osaka-u.ac.jp/ja/research/2016/20161202_1

    • Related Report
      2016 Annual Research Report

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

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