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Detection of histone modification cross-talk using fluorescence cross-correlation spectroscopy

Research Project

Project/Area Number 24657003
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Genetics/Genome dynamics
Research InstitutionTokyo Institute of Technology (2014)
Osaka University (2012-2013)

Principal Investigator

HIROSHI Kimura  東京工業大学, 生命理工学研究科, 教授 (30241392)

Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsクロマチン / 染色体 / 蛍光相互相関分光法 / エピジェネティクス / 蛋白質翻訳後修飾 / 翻訳後修飾 / 抗体 / 蛍光相関分光法 / ヒストン
Outline of Final Research Achievements

Post-translational histone modification plays an important role in the regulation and maintenance of genome function, such as transcription and DNA repair. It remains unknown, however, how a combination of different modifications is regulated and functions specifically. This study aimed to develop a rapid and sensitive system to detect a combination of modifications using fluorescence cross-correlation spectroscopy. When Alexa488-labeled and Cy5-labeled antibodies are mixed with purified histone complexes, they tended to form aggregations, because the solubilization conditions of labeled antibodies and histones were different.This problem was partly solved by using antigen-binding fragments and nucleosomes, instead of IgG and histones. Additional experiments revealed that a combination of different modifications could be detected by measuring fluorescence resonance enery transfer.

Report

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

    (9 results)

All 2014 2012 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Remarks (7 results)

  • [Journal Article] Evaluation of chemical fluorescent dyes as a protein conjugation partner for live cell imaging.2014

    • Author(s)
      Hayashi-Takanaka Y, Stasevich TJ, Kurumizaka H, Nozaki N, Kimura H
    • Journal Title

      PLoS One

      Volume: 9 Issue: 9 Pages: e106271-e106271

    • DOI

      10.1371/journal.pone.0106271

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Monitoring Post-translational Modification on Endogenous Proteins in Living Cells2012

    • Author(s)
      木村 宏
    • Journal Title

      Seibutsu Butsuri

      Volume: 52 Issue: 5 Pages: 234-235

    • DOI

      10.2142/biophys.52.234

    • NAID

      10030877024

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Remarks] 東京工業大学 木村研究室

    • URL

      http://kimura-lab.bio.titech.ac.jp/

    • Related Report
      2014 Annual Research Report
  • [Remarks] 東京工業大学 生命理工学研究科 木村研究室

    • URL

      http://www.bio.titech.ac.jp/laboratory/kimura-kajikawa/kimura/index.html

    • Related Report
      2014 Annual Research Report
  • [Remarks] 木村グループ

    • URL

      http://www.fbs.osaka-u.ac.jp/labs/hiraoka/kimura/index.html

    • Related Report
      2013 Research-status Report
  • [Remarks] 大阪大学研究者総覧 木村 宏

    • URL

      http://www.dma.jim.osaka-u.ac.jp/view?l=ja&u=7721

    • Related Report
      2013 Research-status Report
  • [Remarks] Google Scholar Hiroshi Kimura

    • URL

      http://scholar.google.co.jp/citations?user=z3cSm10AAAAJ&hl=ja&oi=ao

    • Related Report
      2013 Research-status Report
  • [Remarks] 木村グループHP

    • URL

      http://www.fbs.osaka-u.ac.jp/labs/hiraoka/kimura/index.html

    • Related Report
      2012 Research-status Report
  • [Remarks] 大阪大学生命機能研究科教員基本情報 木村 宏

    • URL

      http://www.dma.jim.osaka-u.ac.jp/view?l=ja&u=7721

    • Related Report
      2012 Research-status Report

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

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