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Effect of modified bases on the three-dimensional structure and physiological activities of yeast tyrosine tRNA.

Research Project

Project/Area Number 22550148
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemistry related to living body
Research InstitutionGifu University

Principal Investigator

NISHIKAWA Kazuya  岐阜大学, 工学部, 教授 (60109262)

Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2010: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords生体認識・機能化学 / 修飾塩基 / 核酸・蛋白質・糖化学 / バイオテクノロジー / 核酸塩基 / 転写後修飾
Research Abstract

Originally, it was planned that two types of chimeric tRNA (T5N3, N5T3) were synthesized by exchanged re-union of 5’-and 3’-half fragments obtained through limited digestion of yeast tRNA^Tyr and tRNA^Tyr transcript and used for the analyses on the relationship between the modified base-content and the stability of three-dimensional structure of tRNA. However, it was turned out to be difficult by technical reasons. Therefore, similar experiments were performed by using the anticodon nuclease VapC which was recently developed in our laboratory and E. colitRNA^Leu. The result of Tm-measurement showed that the heat-stability of tRNAs increased in the order of tRNA^Leu transcript<T5N3<N5T3<native tRNA^Leu, indicating that the modified bases, especially those contained in the 5’-half fragment (N5) have the effectof stabilizing three-dimensional structure of tRNA. However, unexpected result was also obtained that the native form of tRNA^Leu is less stable than the transcript tRNA in the absence of Mg2+. It is interesting in a sense that the modified bases can contribute to both of the stability and instability of the higher-order structure of tRNA depending on the state of interaction with Mg2+.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (6 results)

All 2012 2010

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (2 results) Book (2 results)

  • [Journal Article] Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRNA^Tyr pair by the archaeal tyrosine pair in Escherichia coli for genetic code expansion.2010

    • Author(s)
      F. Iraha, K. Oki, T. Kobayashi, S. Ohno, T. Yokogawa, K. Nishikawa, S. Yokoyama and K. Sakamoto
    • Journal Title

      Nucleic Acids Res

      Volume: 38 Pages: 3682-3691

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRN A^<Tyr> pair by the archaeal tyrosine pair in Escherichia coil for genetic code expansion2010

    • Author(s)
      Fumie Iraha, et al.
    • Journal Title

      Nucleic Acids Research

      Volume: 38 Pages: 3682-3691

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] 好酸好熱古細菌 Sulfolobus tokodaii 由来トキシン-アンチトキシンVap BCのtRNA切断活性.2012

    • Author(s)
      関本文音,松浦由佳,大野敏,西川一八,横川隆志
    • Organizer
      第85回日本生化学会大会
    • Place of Presentation
      福岡国際会議場
    • Year and Date
      2012-12-15
    • Related Report
      2012 Final Research Report
  • [Presentation] 好酸好熱古細菌 Sulfolobus tokodaii 由来トキシン-アンチトキシンVapBCの tRNA切断活性2012

    • Author(s)
      関本文音,松浦由佳,大野敏,西川一八,(代表者:横川隆志)
    • Organizer
      第85回日本生化学会大会
    • Place of Presentation
      福岡国際会議場
    • Related Report
      2012 Annual Research Report
  • [Book] Chapter 19, Incorporation of 3-azidotyrosine into proteins through engineering yeast tyrosyl-tRNA synthetase and its application to site-selective protein modification, in "Cell-Free Protein Production" (Y. Endo et al. eds.)2010

    • Author(s)
      T. Yokogawa, S. Ohno and K. Nishikawa
    • Publisher
      Humana Press
    • Related Report
      2012 Final Research Report
  • [Book] Cell-Free Protein Production-Methods and protocols-, Chapter 192010

    • Author(s)
      Takashi Yokogawa, et al.
    • Publisher
      Humana Press
    • Related Report
      2010 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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