Quantitative real-time in-cell imaging reveals heterogeneous clusters of proteins prior to condensation.

ABSTRACT: Our current understanding of biomolecular condensate formation is largely based on observing the final near-equilibrium condensate state. Despite expectations from classical nucleation theory, pre-critical protein clusters were recently shown to form under subsaturation conditions in vitro; if similar long-lived clusters comprising more than a few molecules are also present in cells, our understanding of the physical basis of biological phase separation may fundamentally change. Here, we combine fluorescence microscopy with photobleaching analysis to quantify the formation of clusters of NELF proteins in living, stressed cells. We categorise small and large clusters based on their dynamics and their response to p38 kinase inhibition. We find a broad distribution of pre-condensate cluster s

SUBMITTER: Lan C 

PROVIDER: S-EPMC10427612 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature