Solutes unmask differences in clustering versus phase separation of FET proteins

Mrityunjoy Kar, Laura T. Vogel, Gaurav Chauhan, Suren Felekyan, Hannes Ausserwöger, Timothy J. Welsh, Furqan Dar, Anjana R. Kamath, Tuomas P. J. Knowles, Anthony A. Hyman (), Claus A. M. Seidel () and Rohit V. Pappu ()
Additional contact information
Mrityunjoy Kar: Max Planck Institute of Cell Biology and Genetics
Laura T. Vogel: Heinrich Heine University
Gaurav Chauhan: Washington University in St. Louis
Suren Felekyan: Heinrich Heine University
Hannes Ausserwöger: University of Cambridge
Timothy J. Welsh: University of Cambridge
Furqan Dar: Washington University in St. Louis
Anjana R. Kamath: Max Planck Institute of Cell Biology and Genetics
Tuomas P. J. Knowles: University of Cambridge
Anthony A. Hyman: Max Planck Institute of Cell Biology and Genetics
Claus A. M. Seidel: Heinrich Heine University
Rohit V. Pappu: Washington University in St. Louis

Nature Communications, 2024, vol. 15, issue 1, 1-21

Abstract: Abstract Phase separation and percolation contribute to phase transitions of multivalent macromolecules. Contributions of percolation are evident through the viscoelasticity of condensates and through the formation of heterogeneous distributions of nano- and mesoscale pre-percolation clusters in sub-saturated solutions. Here, we show that clusters formed in sub-saturated solutions of FET (FUS-EWSR1-TAF15) proteins are affected differently by glutamate versus chloride. These differences on the nanoscale, gleaned using a suite of methods deployed across a wide range of protein concentrations, are prevalent and can be unmasked even though the driving forces for phase separation remain unchanged in glutamate versus chloride. Strikingly, differences in anion-mediated interactions that drive clustering saturate on the micron-scale. Beyond this length scale the system separates into coexisting phases. Overall, we find that sequence-encoded interactions, mediated by solution components, make synergistic and distinct contributions to the formation of pre-percolation clusters in sub-saturated solutions, and to the driving forces for phase separation.

Date: 2024
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DOI: 10.1038/s41467-024-48775-3

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