The emerging concept of phase separation in cell biology postulates that, in concert with the classical structural view of stoichiometric quaternary complexes, ribonucleoproteins (RNPs) and nucleic acids are intricately involved in the formation of meso-scale membraneless compartments, commonly termed RNP granules. Elucidating the structural basis underlying such non-deterministic assemblies remains challenging due to their inherent compositional heterogeneity, conformational flexibility and biophysical dynamics. Techniques which allow characterization of their molecular architectures, ideally within the native cellular environment, are therefore required. We review advances in in situ methods that can provide data on the molecular organization in RNP granules from the level of amino-acid interactions, up to their complex cross-talk in cells. These data combined into integrative modeling and multi-scale simulation frameworks will provide an understanding of RNP granules on a continuum scale of spatial resolutions to elucidate their structure-dynamics-function relations.

细胞生物学中相分离的新兴概念假定，与化学计量四元复合物的经典结构观点一致，核糖核蛋白 (RNP) 和核酸错综复杂地参与了中尺度无膜区室的形成，通常称为 RNP 颗粒。由于其固有的组成异质性、构象灵活性和生物物理动力学，阐明这种非确定性组装背后的结构基础仍然具有挑战性。因此，需要能够表征其分子结构的技术，理想情况下在天然细胞环境中。我们就地审查进展可以提供从氨基酸相互作用水平到它们在细胞中的复杂串扰的 RNP 颗粒中分子组织数据的方法。这些数据结合到综合建模和多尺度模拟框架中，将在空间分辨率的连续尺度上提供对 RNP 颗粒的理解，以阐明它们的结构-动力学-功能关系。