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Study on a switching mechanism in RNA recognition by 3'-5' RNA polymerase

Research Project

Project/Area Number 16K18511
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionNational Institute of Advanced Industrial Science and Technology

Principal Investigator

NAKAMURA Akiyoshi  国立研究開発法人産業技術総合研究所, 生命工学領域, 研究員 (10583891)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
KeywordsX線結晶構造解析 / tRNA修飾 / アミノアシルtRNA合成酵素 / DNA/RNAポリメラーゼ / 核酸 / タンパク質 / 鋳型依存伸長反応 / 酵素 / タンパク質-核酸相互作用 / tRNA
Outline of Final Research Achievements

The tRNAHis guanylyltransferase (Thg1) is a unique enzyme which catalyzes a reverse (3'-5' direction) polymerization. It is expected that the reverse polymerization activity of Thg1 is applied to a novel nucleotide modification or elongation technologies. In this study, we successfully constructed a two-stranded tRNA molecule composed of a primer and a template strand by dividing the parent tRNAHis. It allows Thg1 to incorporate a nucleotide triphosphate onto the 5′-end of single-stranded RNA in the 3′-5′ direction. Furthermore, we demonstrated that human Thg1 catalyzes the nucleotide addition reaction for both human cytosolic tRNAHis and mitochondrial tRNAHis.

Academic Significance and Societal Importance of the Research Achievements

核酸を伸長する酵素(ポリメラーゼ)は、生体外においても容易に核酸の複製や修飾ができることから、遺伝子の解析や組換えなどの技術に幅広く活用されており、生命科学の発展に欠かせないものとなっている。ポリメラーゼの1種であるThg1は自然界で唯一他のポリメラーゼとは逆方向に核酸を伸長する能力があることから、これまでにない遺伝子解析や組み換え技術開発への応用が期待されている。本研究ではThg1の機能解析から、Thg1をより多様な核酸に適用できる可能性を見出した。本研究成果を元に、今後Thg1を利用した新たな核酸合成・修飾技術開発が期待される。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (6 results)

All 2019 2018 2017 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (3 results) Remarks (1 results)

  • [Journal Article] Biochemical analysis of human tRNAHis guanylyltransferase in mitochondrial tRNAHis maturation2018

    • Author(s)
      Nakamura Akiyoshi、Wang Daole、Komatsu Yasuo
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 503 Issue: 3 Pages: 2015-2021

    • DOI

      10.1016/j.bbrc.2018.07.150

    • NAID

      120006718707

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Molecular mechanism of substrate recognition and specificity of tRNAHis guanylyltransferase during nucleotide addition in the 3′-5′ direction2018

    • Author(s)
      Nakamura Akiyoshi、Wang Daole、Komatsu Yasuo
    • Journal Title

      RNA

      Volume: 24 Issue: 11 Pages: 1583-1593

    • DOI

      10.1261/rna.067330.118

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] ヒト由来tRNAHis guanylyltransferaseの立体構造解析2019

    • Author(s)
      中村彰良
    • Organizer
      2018年度量子ビームサイエンスフェスタ
    • Related Report
      2018 Annual Research Report
  • [Presentation] Human tRNAHis guanylyltransferase の機能解析2018

    • Author(s)
      中村彰良, 汪道楽, 小松康雄
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 3′-5′方向塩基伸長を触媒するtRNA修飾酵素の基質特異性解析2017

    • Author(s)
      中村彰良、汪道楽、小松康雄
    • Organizer
      日本薬学会第137年会
    • Place of Presentation
      東北大学川内キャンパス (仙台市)
    • Year and Date
      2017-03-24
    • Related Report
      2016 Research-status Report
  • [Remarks] 産業技術総合研究所 生物プロセス研究部門 生体分子工学研究グループホームページ

    • URL

      https://unit.aist.go.jp/bpri/bpri-bimo/index.html

    • Related Report
      2018 Annual Research Report

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Published: 2016-04-21   Modified: 2020-03-30  

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