Development of intelligent ribozyme and aptamer whose activities switch on in response to environment

• Project/Area Number: 26650014
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Kyoto University
• Principal Investigator: Katahira Masato 京都大学, エネルギー理工学研究所, 教授 (70211844)
• Research Collaborator: NAGATA Takashi 京都大学, エネルギー理工学研究所, 准教授 (10415250) ; MASHIMA Tsukasa 京都大学, エネルギー理工学研究所, 助教 (20707426) ; YAMAOKI Yuudai 京都大学, エネルギー理工学研究所, 博士研究員 (00778095)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: RNA酵素 / カリウムイオン / Ｉｎ ｃｅｌｌ ＮＭＲ / 立体構造 / 4重鎖 / RNAアプタマー / Tatタンパク質 / In cell NMR / リボザイム / ４重鎖 / アプタマー

Outline of Final Research Achievements

Two subunits of ribozyme were joined to each end of rGGAGGAGGAGGA(R12). Under low potassium conditions, R12 takes on an extended structure and thus two subunits are located separately, resulting in no enzymatic activity. Under physiological potassium concentrations, R12 forms a compact quadruplex and thus two subunits are closely located, resulting in exertion of the enzymatic activity. By introducing the complementary strand to ribozyme, the ratio of the activity with/without potassium ions was drastically improved. We also succeeded to directly observe NMR signals of nucleic acids in human cells for the first time.

Research Products

• [Journal Article] K+-responsive off-to-on switching of hammerhead ribozyme through dual G-quadruplex formation requiring no heating and cooling treatment (2015)
  • Author(s): Yamaoki Y, Nagata T, Mashima T, Katahira M
  • Journal Title: Biochemical and biophysical research communications Volume: 468(1-2) Issue: 1-2 Pages: 27-31
  • DOI: 10.1016/j.bbrc.2015.10.173
  • NAID: 120005712951
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Boosting of activity enhancement of K+-responsive quadruplex hammerhead ribozyme (2015)
  • Author(s): Yudai Yamaoki, Tsukasa Mashima, Takashi Nagata and Masato Katahira
  • Journal Title: Chemical Communications Volume: 51 Issue: 27 Pages: 5898-5901
  • DOI: 10.1039/c5cc00961h
  • Peer Reviewed / Open Access
• [Journal Article] RNA-directed amino acid coupling as a model reaction for primitive coded translation (2014)
  • Author(s): Harada, K., Aoyama, S., Matsugami, A., Kumar, P.K.R., Katahira, M., Kato, N. and Ohkanda, J.
  • Journal Title: ChemBioChem Volume: 未定 Issue: 6 Pages: 794-798
  • DOI: 10.1002/cbic.201400029
  • Peer Reviewed
• [Presentation] Development of intelligent ribozyme/aptamer that sense K＋ and switch on their activities (2015)
  • Author(s): 片平正人
  • Organizer: 第５３回日本生物物理学会年会
  • Place of Presentation: 金沢大学（金沢市）
  • Year and Date: 2015-09-13
• [Presentation] Real-time monitoring of enzymatic reaction and switching of ribozyme/aptamer activities through sensing of K+ (2015)
  • Author(s): Katahira, M.
  • Organizer: The 6th Asia-Pacific NMR Symposium
  • Place of Presentation: Hong Kong Univ. Sci. Tech.（中国・香港）
  • Year and Date: 2015-08-13
  • Int'l Joint Research / Invited
• [Presentation] Switching the activity of functional RNA in response to K+ and quantitative NMR (2014)
  • Author(s): Katahira, M.
  • Organizer: The 11th Japan-Korea Bilateral Symposium on Biological NMR
  • Place of Presentation: 大阪大学（吹田市）
  • Year and Date: 2014-12-19
  • Invited
• [Remarks] 片平研究室
  • URL: http://www.iae.kyoto-u.ac.jp/bio/