Analysis of the regulatory system for tRNA gene expression to establish a proper tRNA repertoire.

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06490
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43010:Molecular biology-related
• Research Institution: University of Hyogo
• Principal Investigator: Yoshihisa Tohru 兵庫県立大学, 理学研究科, 教授 (60212312)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: tRNA / 転写制御 / 転写因子 / tRNA検出因子 / tRNAレパートリー

Outline of Final Research Achievements

Accumulating pieces of evidence revealed that tRNAs, essential for translation, is under transcriptional regulation, but their regulatory molecular mechanism has been obscure. To analyze this problem using yeast molecular biological technique, we tried to construct ways for analyze promoter activity of tRNA genes, or tDNAs. First, we adopted RT-qPCR to measure pre-tRNAs with an intron, a primary approximation of transcriptional activity of the corresponding tDNA. Second, we constructed RNAi system where an shRNA against EGFP mRNA is expressed from the tDNA promoter, and tested whether extent of suppression of EGFP expression is correlated with tRNA expression. However, all of our constructs so strongly suppressed EGFP expression that promoter activity of various tDNAs could not be compared.

Free Research Field

分子生物学、細胞生物学

Academic Significance and Societal Importance of the Research Achievements

本研究は今まで知られていなかった個別tRNA遺伝子の発現制御を解析するための研究基盤の構築を試みた。学術的には、生理条件に応じた個別のtRNA遺伝子の発現制御を検出することはできたが、本来の目的である遺伝学的なスクリーニングに耐えるプロモータ活性の測定計の構築には至らなかった。RNAiやCRISPRiに用いられる低分子non-coding RNAの発現をベースとした検出系では、レポーターmRNAの発現量をかなり正確に調製する必要がある。今後は、tDNAプロモータで転写される機能性RNAが直接検出できる系での検討が必要であろう。