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2022 Fiscal Year Final Research Report

Study on substrate tRNA recognition mechanisms of multiple subunit tRNA modification enzymes

Research Project

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Project/Area Number 20H03211
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43030:Functional biochemistry-related
Research InstitutionEhime University

Principal Investigator

Hori Hiroyuki  愛媛大学, 理工学研究科(工学系), 教授 (20256960)

Co-Investigator(Kenkyū-buntansha) 平田 章  徳島大学, 大学院社会産業理工学研究部(理工学域), 准教授 (60527381)
横川 隆志  岐阜大学, 工学部, 教授 (90242304)
Project Period (FY) 2020-04-01 – 2023-03-31
Keywords酵素 / 核酸 / タンパク質 / RNA
Outline of Final Research Achievements

The purpose of this study is the clarification of tRNA recognition mechanisms of multiple subunit tRNA modification enzymes. Eukaryotic tRNA methyltransferase complex (Trm11-Trm112 complex) methylates only 20 tRNA molecular species among 43 tRNA species. We have clarified the recognition sites in tRNA of Trm11-Trm112 complex, namely the G10-C25 base pair, CCA terminus, regular size (5 nt) valiable region, and phsphate-ribose backbone around position 38 in tRNA. Furthermore, we have developed a new 4-thiouridine detection system and a new analysis method for tRNA modification enzymes using a next generation DNA sequencer. Moreover, the procols for the later method have been described in Methods in Enxymology. In addition, the knowledges concering THUMP domain has been summarized in a review.

Free Research Field

機能生物化学

Academic Significance and Societal Importance of the Research Achievements

21世紀に入り、tRNAの修飾ヌクレオシドと修飾酵素の欠損が50種類以上のヒト遺伝病と関係することが判ってきた。したがって、当該領域は、遺伝病の病態の理解、新たな遺伝子診断法や遺伝子治療法の開発に直結する分野となっている。また、tRNA中の修飾ヌクレオシドや修飾酵素は、コレラ菌、チフス菌、レトロウイルスなどの感染必須因子である。ゆえに、tRNAの修飾ヌクレオシドと修飾酵素の研究は感染性微生物防除の観点からも重要である。複合tRNA修飾酵素は精製酵素による解析が難しく、未解明な問題が多い分野であったが、本研究では複合tRNAメチル化酵素の基質tRNA認識機構を明らかにすることができた。

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Published: 2024-01-30  

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