In-cell NMR analysis on the structural stability and interaction of nucleic acids in living human cells

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K16054
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Kyoto University
• Principal Investigator: Yamaoki Yudai 京都大学, エネルギー理工学研究所, 助教 (00778095)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: in-cell NMR / 核酸 / 三重鎖構造 / グアニン四重鎖構造 / ダイナミクス / 相互作用

Outline of Final Research Achievements

The intracellular environment is highly crowded with macromolecules and physicochemical properties inside the cells are different from those of in vitro. Therefore, it has long been thought that structures, dynamics, and interactions of nucleic acids can be different in in-cell and in vitro conditions. Here we evaluated the structural properties of nucleic acids in living human cells by in-cell NMR. In terms of intracellular structure, the direct evidence of the formation of parallel and antiparallel triplex structures were provided. Furthermore, base-pair opening dynamics of telomere G-quadruplex structure in living human cells was evaluated at base-pair resolution. We found that all G-G base-pairs open more frequently in living human cells than in vitro. We also detected the interaction of RNA aptamer to its target small molecule in living human cells. The potential of in-cell NMR for investigation of structural properties of nucleic acids in living human cells was demonstrated.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

本研究では、in-cell NMR法により、ヒト生細胞内において核酸の構造、ダイナミクス、相互作用を解析する手法を確立し、その有用性を実証した。核酸が実際に機能を発揮する細胞内において核酸の構造情報を高い空間分解能で解析可能にする本技術は、核酸が関わる多くの基本的な生命現象の理解に貢献すると期待される。