Application of an effective post-labeling method using RNA methyltransferases and synthetic AdoMet-derivatives.
| Project/Area Number |
21603018
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemical biology
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
BESSHO Yoshitaka 独立行政法人理化学研究所, 機能解析第2研究チーム, チームリーダー (70242815)
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| Research Collaborator |
KAWANO Yoshiaki 独立行政法人理化学研究所, 生命系放射光利用システム開発ユニット, 専任技師 (70321750)
SAULIUS Klimasauskas Institute of Biotechnology, Vilnius University, LITHUANIA, Professor
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2009: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 生物・生体工学 / バイオテクノロジー / 酵素反応 / タンパク質 / メチル基転位酵素 / tRNA / 人工補酵素 / S-アデノシルメチオニン |
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| Research Abstract |
We utilized a chemical biology approach to develop a site-specific RNA labeling method, in which nucleic acids can be covalently modified at nucleotide precision using engineered methyltransferases and synthetic analogs of their cofactor, AdoMet. By conjugating fluorescent probes to the extended group of the synthetic AdoMet, we successfully developed the core technologies to incorporate fluorescent labels into microcrystals of complexes of tRNA and its maturation enzymes, which cannot be observed by visible light, and the instrument to detect the fluorescent label incorporated into the sample crystal. A microcrystal X-ray diffraction method was developed, using the fluorescent microcrystals of the complex of the post-labeled tRNA and its maturation enzymes and the co-axial microscope installed on the micro-focus beamline BL32XU of the SPring-8 synchrotron.
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Report
(4 results)
Research Products
(24 results)
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[Journal Article] Crystal structure of Methanocaldococcus jannaschii Trm4 complexed with sinefungin2010
Author(s)
Kuratani M, Hirano M, Goto-Ito S, Itoh Y, Hikida Y, Nishimoto M, Sekine S, Bessho Y, Ito T, Grosjean H, and Yokoyama S
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Journal Title
J Mol Biol
Volume: 401(3)
Pages: 323-333
Related Report
Peer Reviewed
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[Presentation] Multisite specific tRNA methyltransferase Trm1 from a hyper-thermophilic eubacterium, Aquifex aeolicus2011
Author(s)
Takako Awai, Anna Ochi, Ihsanawati, Toru Sengoku, Satoshi Kimura, Takashi Yokogawa, Akira Hirata, Yoshitaka Bessho, Tsutomu Suzuki, Shigeyuki Yokoyama, Hiroyuki Hori
Organizer
Thermophiles 2011
Place of Presentation
Montana USA
Year and Date
2011-09-11
Related Report
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