Cell culture conditions regulates histone modification.

• Project/Area Number: 25503002
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: epigenetics
• Research Institution: Tokyo Institute of Technology (2014-2015); Osaka University (2013)
• Principal Investigator: Hayashi-Takanaka Yoko 東京工業大学, 生命理工学研究科, 産学官連携研究員 (50397551)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ヒストン修飾 / 細胞培養 / メチル化 / アセチル化

Outline of Final Research Achievements

The levels of histone modifications change depending on the surrounding environments. To validate the effects of cell culture condition on histone modification, cells were cultured under various cell conditions and the levels of histone modifications were measured.
When dense cell cultured supernatant were added to the culture medium, the levels of histone acetylation, and methylation, increased and decreased, respectively. The levels of histone acetylation changed within 3 hr, which was relatively fast. On the other hands, the levels of methylation decreased after 24 hr, suggesting that the histone acetylation and methylation are regulated by different mechanism. Furthermore, we found that one of the factor which changed histone modifications in dense cell cultured supernatant was lactic acid.

Research Products

• [Journal Article] Histone H4 lysine 20 acetylation is associated with gene repression in human cells. (2016)
  • Author(s): Kaimori, J. Y., Maehara, K., Hayashi-Takanaka, Y., Harada, A., Fukuda, M., Yamamoto, S., Ichimaru, N., Umehara, T., Yokoyama, S., Matsuda, R., Ikura, T., Nagao, K., Obuse, C., Nozaki, N., Takahara, S., Takao, T., Ohkawa, Y., Kimura, H., and Isaka, Y.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 24318-24318
  • DOI: 10.1038/srep24318
  • NAID: 120005770760
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies. (2015)
  • Author(s): Hayashi-Takanaka Y., Maehara K., Harada A., Umehara T., Yokoyama S., Obuse C., Ohkawa Y., Nozaki N., Kimura H.
  • Journal Title: Chromosome Res. Volume: 23 Issue: 4 Pages: 753-766
  • DOI: 10.1007/s10577-015-9486-4
  • Peer Reviewed / Open Access
• [Journal Article] Visualizing posttranslational and epigenetic modifications of endogenous proteins in vivo. (2015)
  • Author(s): Kimura H, Hayashi-Takanaka Y, Stasevich TJ, Sato Y
  • Journal Title: Histochem Cell Biol Volume: 144 Issue: 2 Pages: 101-109
  • DOI: 10.1007/s00418-015-1344-0
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Early Development of Cloned Bovine Embryos Produced from Oocytes Enucleated by Fluorescence Metaphase II Imaging Using a Conventional Halogen-Lamp Microscope. (2015)
  • Author(s): .Iwamoto D, Yamagata K, Kishi M, Hayashi-Takanaka Y, Kimura H, Wakayama T, Saeki K.
  • Journal Title: Cell Reprogram Volume: 17 Issue: 2 Pages: 106-114
  • DOI: 10.1089/cell.2014.0086
  • Peer Reviewed
• [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] 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
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural polymorphism in the L1 loop regions of human H2A. Z. 1 and H2A. Z. 2. (2013)
  • Author(s): Naoki Horikoshi, Koichi Sato, Keisuke Shimada, Yasuhiro Arimura, Akihisa Osakabe, Hiroaki Tachiwana, Yoko Hayashi-Takanaka, Wakana Iwasaki, Wataru Kagawa, Masahiko Harata, Hiroshi Kimura and Hitoshi Kurumizaka.
  • Journal Title: Acta Crystallographica Section D Volume: 69 Issue: 12 Pages: 2431-2439
  • DOI: 10.1107/s090744491302252x
  • Peer Reviewed
• [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
• [Presentation] 特異的抗体によるヒストンH4修飾動態の単一細胞解析 (2015)
  • Author(s): 林陽子・野崎直仁・木村宏
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Distribution and its function of histone H4 modifications revealed by a panel of specific monoclonal antibodies (2015)
  • Author(s): Hayashi-Takanaka Yoko, Naohito Nozaki, Hiroshi Kimura
  • Organizer: 内藤カンファレンス(Epigenetics-From histone code to therapeutic strategy)
  • Place of Presentation: 札幌
  • Year and Date: 2015-09-16
  • Int'l Joint Research
• [Presentation] 多重免疫染色法を用いた細胞周期や代謝産物によるヒストン修飾動態の計測 (2014)
  • Author(s): 林陽子・野崎直仁・木村宏
  • Organizer: がん代謝研究会
  • Place of Presentation: 東京理科大学
  • Year and Date: 2014-07-10 – 2014-07-11
• [Presentation] 細胞周期におけるヒストン修飾動態の検出 (2013)
  • Author(s): 林陽子・野崎直仁・木村宏
  • Organizer: 定量生物学会
  • Place of Presentation: 大阪大学
• [Presentation] 生細胞における蛍光化学物質の評価 (2013)
  • Author(s): 林陽子・越阪部晃永・スタセビッチティモシー・加藤大貴・海沼嵩・胡桃坂仁志・野崎直仁・木村宏
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド