Solutes unmask differences in clustering versus phase separation of FET proteins.
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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.
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Acknowledgements: We are grateful to Andrei Pozniakovsky for the DNA constructs of all proteins, to Régis Lemaitre and Barbara Borgonovo for technical support with protein expression, purification, and characterization at MPI-CBG, and to Ralf Kühnemuth and Oleg Opanasyuk for technical assistance. We thank Eric Geertsma, Timothy Lohman, and Min Kyung Shinn for helpful discussions. This work was funded by a direct grant from the Max Planck Society (to A.A.H.), a grant from the NOMIS foundation (to A.A.H.), the Wellcome trust (209194/Z/17/Z to A.A.H.), the Deutsche Forschungsgemeinschaft (DFG) under Germany’s Excellence Strategy—EXC-2068—390729961- Cluster of Excellence Physics of Life of TU Dresden (to A.A.H.), the European Research Council through the ERC grant PhysProt (to T.P.J.K., agreement no. 337969), the Wellcome Trust and the Frances and Augustus Newman foundation (to T.P.J.K.), SPP2191 from the Deutsche Forschungsgemeinschaft (to C.A.M.S. and R.V.P.), the US National Institutes of Health (R01NS121114 to R.V.P.), the US National Science Foundation (MCB-2227268 to R.V.P.), and the St. Jude Children’s Research Hospital collaborative research consortium on the Biology and Biophysics of RNP Granules (to R.V.P.).
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2041-1723
