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

Published version
Peer-reviewed

Repository DOI


Type

Article

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Authors

Lan, Chenyang 
Kim, Juhyeong 
Aprile-Garcia, Fernando  ORCID logo  https://orcid.org/0000-0002-5133-0857
Weyrauch, Sophie 

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.

Description

Keywords

Proteins, Cognition, Diagnostic Imaging

Journal Title

Nat Commun

Conference Name

Journal ISSN

2041-1723
2041-1723

Volume Title

Publisher

Springer Science and Business Media LLC
Sponsorship
This work was supported by the European Research Council (grant agreement No. 681891) and the Deutsche Forschungsgemeinschaft (DFG) under Germany's Excellence Strategy (CIBSS EXC-2189 Project ID 390939984) and the SFB1381 programme (Project ID 403222702).
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