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Structural study of the higher order chromatin structure formed by HP1BP3

Research Project

Project/Area Number 15H06692
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Structural biochemistry
Research InstitutionWaseda University

Principal Investigator

Koyama Masako  早稲田大学, 理工学術院, 次席研究員(研究院助教) (40755097)

Project Period (FY) 2015-08-28 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsクロマチン / ヌクレオソーム / 分裂酵母 / リンカーヒストン / ヒストン / HP1BP3 / X線結晶解析
Outline of Final Research Achievements

We have aimed to determine the structure of the fundamental unit of higher order chromatin formed by human HP1BP3, the recently identified linker histone in human. However, it was difficult to purify the HP1BP3 protein at high yield necessary for the structural studies. Therefore, we selected the functional homolog of HP1BP3 in S. pombe. The protein was expressed as a soluble recombinant protein in E. coli, and purified in high quality. Furthermore, we established the method to reconstitute S. pombe nucleosomes in vitro. We purified the four canonical core histones in S. pombe (H2A, H2B, H3, and H4, which share 79%, 68%, 92%, and 91% amino acid sequence homology with their human counterparts). Using S. pombe histones and a 146-base-pair DNA, we successfully reconstituted nucleosomes in vitro. We confirmed that the HP1BP3 homolog in S. pombe bind the nucleosome.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Annual Research Report
  • Research Products

    (5 results)

All 2017 2016

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Acknowledgement Compliant: 1 results) Presentation (4 results)

  • [Journal Article] In vitro reconstitution and biochemical analyses of theSchizosaccharomyces pombe nucleosome.2017

    • Author(s)
      Masako Koyama, Wataru Nagakura, Hiroki Tanaka, Tomoya Kujirai, Yuji Chikashige, Tokuko Haraguchi, Yasushi Hiraoka, Hitoshi Kurumizaka.
    • Journal Title

      Biochemical and biophysical research communications.

      Volume: 482 Issue: 4 Pages: 896-901

    • DOI

      10.1016/j.bbrc.2016.11.130

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Oct4 が標的ヌクレオソームを特異的に認識する機構の解明2017

    • Author(s)
      小山昌子、水上優夏、胡桃坂仁志
    • Organizer
      第34回染色体ワークショップ・第15回核ダイナミクス研究会
    • Place of Presentation
      千葉県木更津市
    • Year and Date
      2017-01-11
    • Related Report
      2016 Annual Research Report
  • [Presentation] Biochemical and biophysical analyses of the S. pombe nucleosome2017

    • Author(s)
      Masako Koyama
    • Organizer
      Japan-Swiss Symposium "Chromatin Structure and Dynamics"
    • Place of Presentation
      スイス、バーゼル
    • Related Report
      2016 Annual Research Report
  • [Presentation] 細胞初期化に重要なパイオニア転写因子による 標的ヌクレオソーム認識機構の解明2016

    • Author(s)
      小山昌子、胡桃坂仁志
    • Organizer
      第39回 日本分子生物学会年会
    • Place of Presentation
      神奈川県横浜市
    • Year and Date
      2016-11-30
    • Related Report
      2016 Annual Research Report
  • [Presentation] パイオニア転写因子Oct4による標的ヌクレオソーム認識機構の解明2016

    • Author(s)
      小山昌子、胡桃坂仁志
    • Organizer
      新学術領域研究班会議「クロマチン動構造ワークショップ」
    • Place of Presentation
      北海道虻田郡
    • Year and Date
      2016-07-07
    • Related Report
      2016 Annual Research Report

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Published: 2015-08-26   Modified: 2018-03-22  

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