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Reconstitution of mitotic chromatids with a minimum set of purified proteins

Research Project

Project/Area Number 24770172
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Molecular biology
Research InstitutionThe Institute of Physical and Chemical Research

Principal Investigator

KEISHI Shintomi  独立行政法人理化学研究所, 平野染色体ダイナミクス研究室, 研究員 (60462694)

Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Keywords染色体 / コンデンシン / トポイソメラーゼⅡ / ヒストン / ヒストンシャペロン / 再構成 / 分裂期 / 細胞周期 / トポⅡ / トポII / リン酸化
Outline of Final Research Achievements

Mitotic chromatids assembly helps guarantee error-free transmission of the genome during cell division. To understand mechanistically how chromatids might be assembled, I have aimed to reconstitute them with as few purified proteins as possible in a test tube. Intensive surveys on ingredients of the assay, which are based on the hypothesis that proper targeting of major chromosomal proteins to sperm DNA is the key for the reconstitution, enable me to recapitulate the chromatid assembly process using only six protein factors (core histones, topoisomerase II, condensin I, nucleoplasmin, Nap1, FACT) and ATP. These results indicate that chromatid assembly during mitosis hinges on much simpler mechanisms than previously thought. Equally important, the chromatid reconstitution system established in this study will undoubtedly open a new avenue of dissecting specific functions of individual factors as well as their cooperation in the future.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • Research Products

    (8 results)

All 2015 2014 Other

All Presentation (8 results) (of which Invited: 2 results)

  • [Presentation] Reconstitution of mitotic chromatids with a minimum set of purified factors2015

    • Author(s)
      Keishi Shintomi
    • Organizer
      EMBO workshop - SMC proteins
    • Place of Presentation
      Vienna, Austria
    • Year and Date
      2015-05-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] Reconstitution of mitotic chromatids with a minimum set of purified components2014

    • Author(s)
      Keishi Shintomi
    • Organizer
      The 9th 3R symposium
    • Place of Presentation
      Gotemba, Japan
    • Year and Date
      2014-11-19
    • Related Report
      2014 Annual Research Report
  • [Presentation] Can we reconstitute a mitotic chromosome from purified components?

    • Author(s)
      Keishi Shintomi
    • Organizer
      熊本大学リエゾンラボ研究会・HIGO プログラム最先端研究セミナー
    • Place of Presentation
      熊本県熊本市
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] 受精卵における父性ゲノム動態の試験管内再構成

    • Author(s)
      新冨圭史、平野達也
    • Organizer
      第65回日本細胞生物学会
    • Place of Presentation
      愛知県名古屋市
    • Related Report
      2013 Research-status Report
  • [Presentation] 精製タンパク質を用いた分裂期染色体の再構成

    • Author(s)
      新冨圭史、平野達也
    • Organizer
      第31回染色体ワークショップ
    • Place of Presentation
      神奈川県箱根町
    • Related Report
      2013 Research-status Report
  • [Presentation] 精製タンパク質を用いて染色体を作ることができるか?

    • Author(s)
      新冨圭史、平野達也
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場(福岡)
    • Related Report
      2012 Research-status Report
  • [Presentation] Can we reconstitute a chromosome from purified components?

    • Author(s)
      新冨圭史
    • Organizer
      熊本大学リエゾンラボ研究会リーディングプログラム:HIGO最先端研究セミナー
    • Place of Presentation
      熊本大学発生医学研究所(熊本)
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] 受精卵における父性ゲノム動態の試験管内再構成

    • Author(s)
      新冨圭史、平野達也
    • Organizer
      第65回日本細胞生物学会大会
    • Place of Presentation
      ウインクあいち(名古屋)
    • Related Report
      2012 Research-status Report

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

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