Program in Cellular Molecular Developmental Biology and Biophysics, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218, USA.
Program in Cellular Molecular Developmental Biology and Biophysics, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218, USA; Department of Biophysics, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218, USA.
Curr Opin Chem Biol. 2023 Oct;76:102371. doi: 10.1016/j.cbpa.2023.102371. Epub 2023 Jul 29.
Protein-protein and protein-RNA interactions are essential for cell function and survival. These interactions facilitate the formation of ribonucleoprotein complexes and biomolecular condensates via phase separation. Such assembly is involved in transcription, splicing, translation and stress response. When dysregulated, proteins and RNA can undergo irreversible aggregation which can be cytotoxic and pathogenic. Despite technical advances in investigating biomolecular condensates, achieving the necessary spatiotemporal resolution to deduce the parameters that govern their assembly and behavior has been challenging. Many laboratories have applied advanced microscopy methods for imaging condensates. For example, single molecule imaging methods have enabled the detection of RNA-protein interaction, protein-protein interaction, protein conformational dynamics, and diffusional motion of molecules that report on the intrinsic molecular interactions underlying liquid-liquid phase separation. This review will outline advances in both microscopy and spectroscopy techniques which allow single molecule detection and imaging, and how these techniques can be used to probe unique aspects of biomolecular condensates.
蛋白质-蛋白质和蛋白质-RNA 相互作用对于细胞功能和存活至关重要。这些相互作用通过相分离促进核糖核蛋白复合物和生物分子凝聚体的形成。这种组装涉及转录、剪接、翻译和应激反应。当失调时，蛋白质和 RNA 可能会发生不可逆转的聚集，这可能是细胞毒性和致病性的。尽管在研究生物分子凝聚体方面取得了技术进步，但要实现推断控制其组装和行为的参数所需的时空分辨率一直具有挑战性。许多实验室已经应用了先进的显微镜方法来对凝聚体进行成像。例如，单分子成像方法能够检测 RNA-蛋白质相互作用、蛋白质-蛋白质相互作用、蛋白质构象动力学以及报告液-液相分离下内在分子相互作用的分子扩散运动。本综述将概述允许单分子检测和成像的显微镜和光谱技术的进展，以及如何使用这些技术来探测生物分子凝聚体的独特方面。
