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Development of a new functional analysis using mutational profiling that reveals RNA-protein interaction

Research Project

Project/Area Number 21K20638
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeMulti-year Fund
Review Section 0701:Biology at molecular to cellular levels, and related fields
Research InstitutionEhime University

Principal Investigator

Yamagami Ryota  愛媛大学, 理工学研究科(工学系), 助教 (70767227)

Project Period (FY) 2021-08-30 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
KeywordsRNA / tRNA / tRNA修飾 / tRNA修飾酵素 / 網羅解析 / 変異プロファイリング法 / RNAメチル化酵素 / 次世代シーケンス / 変異プロファイリング / RNAシーケンス / 核酸関連タンパク質
Outline of Research at the Start

近年の研究より、RNAの配列や構造因子は、多くの遺伝子の発現を調節することが明らかになっています。RNAは、転写後にプロセシングやRNA修飾などの様々な成熟過程を経て、その機能性を獲得します。その一方で、「RNAの配列と構造がどのようにしてRNA成熟化酵素と相互作用し、どのようにRNA成熟化機構の制御に関わるのか」に関するメカニズムの全貌は、未解明です。本研究では、RNA修飾酵素やプロセシング酵素と基質RNAとの相互作用をハイスループットに解析する技術を開発します。本技術を用いて、RNAの配列や構造を介したRNA成熟化反応の制御機構とそれによる遺伝子発現調節機構を探ります。

Outline of Final Research Achievements

The objective of this project is to develop a new high-throughput technique for conducting functional analyses of RNA modification enzymes. The findings have been reported as follows: (1) We have developed tRNA-MaP, a technique that selectively detects substrate tRNAs from a mixture of substrate and non-substrate tRNAs. (2) By using tRNA-MaP, we were able to reveal the substrate tRNA recognition mechanism of tRNA m1A22 methyltransferase (TrmK) from Geobacillus stearothermophilus, a thermophilic bacteria. (3) In combination with protein orthology analysis, tRNA-MaP was used to predict the tRNA modification enzyme genes encoded in the G. stearothermophilus genome.

Academic Significance and Societal Importance of the Research Achievements

本研究は、RNAとRNA修飾酵素の相互作用機構とその機能の理解を深めることを目的としています。本研究によって開発されたtRNA-MaP法は、本目的の達成を加速させるものであり、幅広い分野での応用が期待できます。
This project aims to enhance our understanding of RNA modification enzymes and their functions, and the developed tRNA-MaP technique has potential implications for further studies in this field.

Report

(3 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • Research Products

    (7 results)

All 2023 2022 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 1 results) Presentation (1 results) Remarks (4 results)

  • [Journal Article] Application of mutational profiling: New functional analyses reveal the tRNA recognition mechanism of tRNA m1A22 methyltransferase2023

    • Author(s)
      Ryota Yamagami, Hiroyuki Hori
    • Journal Title

      Journal of Biological Chemistry

      Volume: 299 Issue: 1 Pages: 102759-102776

    • DOI

      10.1016/j.jbc.2022.102759

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Functional analysis of tRNA modification enzymes using mutational profiling2023

    • Author(s)
      Ryota Yamagami, Hiroyuki Hori
    • Journal Title

      Methods in Enzymology

      Volume: - Pages: 69-101

    • DOI

      10.1016/bs.mie.2023.02.021

    • ISBN
      9780443239762
    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Presentation] Mutational Profiling Reveals The tRNA Recognition Mechanism of tRNA m1A Methyltransferase (TrmK)2022

    • Author(s)
      Ryota Yamagami, Hiroyuki Hori
    • Organizer
      第23回日本RNA学会年会
    • Related Report
      2022 Annual Research Report
  • [Remarks] tRNA-MaP: functional analyses of RNA-related

    • URL

      https://www.eurekalert.org/news-releases/975353

    • Related Report
      2022 Annual Research Report
  • [Remarks] tRNA-MaP: functional analyses of RNA-related

    • URL

      https://www.alphagalileo.org/en-gb/Item-Display/ItemId/228980?returnurl=https://www.alphagalileo.org/en-gb/Item-Display/ItemId/228980

    • Related Report
      2022 Annual Research Report
  • [Remarks] tRNA-MaP: functional analyses of RNA-related

    • URL

      https://www.asiaresearchnews.com/content/trna-map-functional-analyses-rna-related-enzymes-using-next-generation-dna-sequencer

    • Related Report
      2022 Annual Research Report
  • [Remarks] 次世代DNAシーケンサを併用した新しいRNA関連酵素の解析法(tRNA-MaP)を開発

    • URL

      https://research.ehime-u.ac.jp/post-ja/

    • Related Report
      2022 Annual Research Report

URL: 

Published: 2021-10-22   Modified: 2024-01-30  

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