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 sizes and show that NELF protein cluster formation can be explained as non-classical nucleation with a surprisingly flat free-energy landscape for a wide range of sizes and an inhibition of condensation in unstressed cells

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• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch
• Notes: Nature Communications. - 14, 1 (2023) , 4831, ISSN: 2041-1723

Contributors, Places and Time

• Event: Veröffentlichung
• (where): Freiburg
• (who): Universität
• (when): 2023
• Creator: Lan, Chenyang ; Kim, Juhyeong ; Ulferts, Svenja ; Aprile-Garcia, Fernando ; Weyrauch, Sophie ; Anandamurugan, Abhinaya ; Grosse, Robert ; Sawarkar, Ritwick ; Reinhardt, Aleks ; Hugel, Thorsten

Further information

• DOI: 10.1038/s41467-023-40540-2
• URN: urn:nbn:de:bsz:25-freidok-2389261
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.