Super-resolution single-molecule dynamics analysis of phase separation formation by the development of a drug-inducible system for intracellular phase separation

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K15755
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Ito Yuma 東京工業大学, 生命理工学院, 助教 (70803245)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Keywords: 1分子計測 / 生細胞イメージング / 相分離

Outline of Final Research Achievements

We have established an artificial system to induce intracellular droplet formation by adding drugs to investigate the phase separation mechanism essential for biological functions. This method enables us to measure molecular dynamics in droplets using single-molecule imaging in living cells. We obtained the characteristic dynamics in droplet formation as precise quantitative information at the single-molecule level for the intrinsically disordered regions of various phase-separated proteins. This method quantified the effect of the intracellular environment, molecular diffusion and exchange inside and outside the droplet, and the amino acid composition that contributes to phase separation. This study provides a tool to investigate the physical and biological properties of phase separation, which could contribute widely to life science research.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

細胞内の相分離は遺伝子発現やタンパク質分解などの広範な生体機能を制御しており、がんや神経変性疾患などにも関与している。本研究で開発した相分離誘導系は、液滴形成における特徴的な動態を顕在化し、その基盤となる物理的な性質や分子生物学的な機構を定量的かつ高精細に計測できるものである。さらに生化学等の研究手法にも広く適用できると考えられ、相分離が関与する広範な生命科学研究へ貢献すると期待できる。