Single molecule analysis in the generation of oscillatory circuit for cAMP signaling

• Project/Area Number: 15K14491
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: The University of Tokushima
• Principal Investigator: HORIKAWA Kazuki 徳島大学, 大学院医歯薬学研究部(医学系), 教授 (70420247)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: イメージング / 生物リズム / 自己組織化 / cAMP / 離散系 / 興奮系 / バイオメージング / 一分子計測 / シグナル伝達

Outline of Final Research Achievements

To ask the mechanisms in the construction of oscillatory signaling circuit, we performed single molecule quantification for essential components in the cAMP signaling system. We developed essential technologies including the efficient generation of GFP-tagged cell lines(BMC Biotech, 2016) and ultrasensitive cAMP indicators (ACS Chem Biol, 2016). By utilizing these tools, we performed a simultaneous imaging of cellular function (cAMP) and molecular quantification and identified that self-organized construction of signaling circuit is regulated by few proteins characterized with significantly low level expression.

Research Products

• [Journal Article] Nontrivial Effect of the Color-Exchange of a Donor/Acceptor Pair in the Engineering of Forster Resonance Energy Transfer (FRET)-Based Indicators. (2016)
  • Author(s): Yusaku Ohta, Takanori Kamagata, Asuka Mukai, Shinji Takada, Takeharu Nagai, and Kazuki Horikawa
  • Journal Title: ACS Chemical Biology Volume: 11(7) Issue: 7 Pages: 1816-1822
  • DOI: 10.1021/acschembio.6b00221
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Reliable handling of highly A/T-rich genomic DNA for efficient generation of knockin strains of Dictyostelium discoideum (2016)
  • Author(s): Asuka Mukai, Aya Ichiraku, Kzuki Horikawa
  • Journal Title: BMC Biotechnology Volume: なし Issue: 1 Pages: 37-37
  • DOI: 10.1186/s12896-016-0267-8
  • NAID: 120006814108
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Effective impairment of myeloma cells and their progenitors by blockade of monocarboxylate transportation (2015)
  • Author(s): 6. Hanson DJ, Nakamura S, Amachi R, Hiasa M, Oda A, Tsuji D, Itoh K, Harada T, Horikawa K, Teramachi J, Miki H, Matsumoto T, Abe M.
  • Journal Title: Oncotarget Volume: 6 Issue: 32 Pages: 33568-33586
  • DOI: 10.18632/oncotarget.5598
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Minority control of synchronized dynamics in biological oscillator (2015)
  • Author(s): 堀川一樹
  • Organizer: 第38回日本分子生物学会
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
  • Year and Date: 2015-12-01
• [Presentation] 自発的発振回路の新規構築原理 (2015)
  • Author(s): 堀川一樹
  • Organizer: 日本顕微鏡学会第71回学術講演会
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2015-05-13
• [Book] 少数性生物学 (2017)
  • Author(s): 永井健治、村越秀治、大場雄介、富樫祐一、小松﨑民樹、城口克之、野地博行、前島一博、粟津暁紀、鈴木宏明、茅 元司、矢島潤一郎、谷口雄一、小嶋誠司、今田勝巳、石島秋彦、福岡 創、蔡 栄淑、堀川一樹、大出晃士
  • Total Pages: 178
  • Publisher: 日本評論社