Real-time imaging of histone modification dynamics in the specific gene loci

• Project/Area Number: 25650009
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: Tokyo Institute of Technology (2014-2015); Osaka University (2013)
• Principal Investigator: Sato Yuko 東京工業大学, 生命理工学研究科, 科学研究費研究員 (70435882)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: エピジェネティクス / イメージング / クロマチン / ライブイメージング / 蛍光プローブ / ヒストン修飾 / 細胞内抗体 / scFv / 細胞核 / 遺伝子

Outline of Final Research Achievements

This project aimed to visualize the dynamics of histone modification in the specific gene loci in living cells. To visualize specific genome loci, we searched repetitive DNA sequence enriched in single loci and designed TALE protein. However, TALEs that are expected to bind to these sequences failed to label them in living cells. On the other hand, using a mintbody specific to phosphorylated RNA polymerase II combined with ANCHOR system that labeled the specific locus, active transcription at the single gene locus was visualized in human cells.

Research Products

• [Int'l Joint Research] トゥールーズ第三大学(フランス)
• [Journal Article] Histone Acetylation on <i>Drosophila</i> Polytene Chromosomes Visualized by Mintbody (2015)
  • Author(s): Sato Y, Mukai M, Kimura H
  • Journal Title: CYTOLOGIA Volume: 80 Issue: 4 Pages: 383-384
  • DOI: 10.1508/cytologia.80.383
  • NAID: 130005118546
  • ISSN: 0011-4545, 1348-7019
  • Open Access
• [Journal Article] Quantifying histone and RNA polymerase II post-translational modification dynamics in mother and daughter cells. (2014)
  • Author(s): Stasevich TJ, Sato Y, Nozaki N, Kimura H.
  • Journal Title: Methods Volume: 70 Issue: 2-3 Pages: 77-88
  • DOI: 10.1016/j.ymeth.2014.08.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Regulation of RNA polymerase II activation by histone acetylation in single living cells (2014)
  • Author(s): Stasevich TJ, Hayashi-Takanaka Y, Sato Y, Maehara K, Ohkawa Y, Sakata-Sogawa K, Tokunaga M, Nagase T, Nozaki N, McNally JG, Kimura H
  • Journal Title: Nature Volume: 516 Issue: 7530 Pages: 272-275
  • DOI: 10.1038/nature13714
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Genetically encoded system to track histone modification in vivo. (2013)
  • Author(s): Yuko Sato, Masanori Mukai, Jun Ueda, Michiko Muraki, Timothy J. Stasevich, Naoki Horikoshi, Tomoya Kujirai, Hiroaki Kita, Taisuke Kimura, Seiji Hira, Yasushi Okada, Yoko Hayashi-Takanaka, Chikashi Obuse, Hitoshi Kurumizaka, Atsuo Kawahara, Kazuo Yamagata, Naohito Nozaki and Hiroshi Kimura
  • Journal Title: Scientific Reports Volume: 3 Issue: 1 Pages: 2436-2436
  • DOI: 10.1038/srep02436
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis of a nucleosome containing histone H2A. B/H2A. Bbd that transiently associates with reorganized chromatin. (2013)
  • Author(s): Arimura Y., Kimura H., Oda T., Sato K., Osakabe A., Tachiwana H., Sato Y., Kinugasa Y., Ikura T., Sugiyama M., Sato M., Kurumizaka H.
  • Journal Title: Sci Rep. Volume: 3 Issue: 1 Pages: 3510-3510
  • DOI: 10.1038/srep03510
  • Peer Reviewed / Open Access
• [Presentation] Visualizing the inactive X chromosome in vivo by using H4K20me1-specific intracellular antibody probe (2015)
  • Author(s): Sato Y, Kimura H
  • Organizer: EMBL Conference “Nuclear structure and dynamics”
  • Place of Presentation: Isle sur Sorgue （フランス）
  • Year and Date: 2015-10-07
  • Int'l Joint Research
• [Presentation] H4K20me1特異的細胞内抗体を用いた不活性X染色体動態のin vivo解析 (2014)
  • Author(s): 佐藤優子、木村宏
  • Organizer: 第37回 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-12-25 – 2014-12-27
• [Presentation] Visualization of histone modification dynamics during zygotic genome activation in Zebrafish (2014)
  • Author(s): Sato Y and Kimura H
  • Organizer: 第20回 小型魚類研究会
  • Place of Presentation: 慶應義塾大学 薬学部（芝共立キャンパス）
  • Year and Date: 2014-09-20 – 2014-09-21
• [Presentation] Intracellular antibody probes for visualizing histone modifications in vivo (2014)
  • Author(s): Sato Y and Kimura H
  • Organizer: 第37回 内藤コンファレンス
  • Place of Presentation: 北海道 ヒルトンニセコビレッジ
  • Year and Date: 2014-07-15 – 2014-07-18
• [Presentation] 細胞内抗体プローブを用いたヒストン修飾動態の生体イメージング (2014)
  • Author(s): 佐藤優子、木村宏
  • Organizer: 第8回 日本エピジェネティクス研究会年会
  • Place of Presentation: 東京大学 伊藤国際学術研究センター
  • Year and Date: 2014-05-25 – 2014-05-27
• [Presentation] Genetically encoded system to track histone acetylation in vivo
  • Author(s): 佐藤 優子, 向 正則, Timothy J. Stasevich，木村 宏
  • Organizer: EMBO Conference Series Chromatin and Epigenetics
  • Place of Presentation: EMBL Heidelberg, Germany
• [Presentation] 細胞内抗体プローブを用いたヒストンアセチル化動態のin vivoイメー
  • Author(s): 佐藤優子、向 正則、Timothy J. Stasevich、木村 宏
  • Organizer: 第７回日本エピジェネティクス研究会年会
  • Place of Presentation: 奈良県新公会堂
• [Presentation] ゼブラフィッシュ初期過程におけるヒストンアセチル化動態のin vivoイメージング
  • Author(s): 佐藤 優子, 村木 倫子, 川原 敦雄, 木村宏
  • Organizer: 第36回 日本分子生物学会年会
  • Place of Presentation: 神戸国際展示場
• [Book] Optical Probes in Biology (Jin Zhang, Sohum Mehta, Carsten Schultz ed) (2014)
  • Author(s): Kimura H and Sato Y
  • Total Pages: 511
  • Publisher: CRC Press, London
• [Book] エピジェネティクス-基礎研究から産業応用への展望- (2014)
  • Author(s): 佐藤 優子, 木村 宏
  • Total Pages: 9
  • Publisher: シーエムシー
• [Book] エピジェネティクスと病気 (2013)
  • Author(s): 木村 宏, 佐藤 優子
  • Total Pages: 7
  • Publisher: メディカルドゥ
• [Remarks] 大阪大学ホームページ
  • URL: http://www.fbs.osaka-u.ac.jp/labs/hiraoka/kimura/index.html
• [Remarks] 大阪大学生命機能研究科の研究成果紹介のページ
  • URL: http://www.fbs.osaka-u.ac.jp/jpn/events/achievement/sato-kimura-20130812/
• [Remarks] 大阪大学の研究成果紹介ページ
  • URL: http://resou.osaka-u.ac.jp/ja/research/2013/20130814_1