Ion binding with charge inversion combined with screening modulates DEAD box helicase phase transitions

Summary

Membraneless organelles, or biomolecular condensates, enable cells to compartmentalize material and processes into unique biochemical environments. While specific, attractive molecular interactions are known to stabilize biomolecular condensates, repulsive interactions, and the balance between these opposing forces, are largely unexplored. Here, we demonstrate that repulsive and attractive electrostatic interactions regulate condensate stability, internal mobility, interfaces, and selective partitioning of molecules both in vitro and in cells. We find that signaling ions, such as calcium, alter repulsions between model Ddx3 and Ddx4 condensate proteins by directly binding to negatively charged amino acid sidechains and effectively inverting their charge, in a manner fundamentally dissimilar to electrostatic screening. Using a polymerization model combined with generalized stickers and spacers, we accurately quantify and predict condensate stability over a wide range of pH, salt concentrations, and amino acid sequences. Our model provides a general quantitative treatment for understanding how charge and ions reversibly control condensate stability.

Graphical abstract

Highlights

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  Multivalent cations promote Ddx4 phase transitions

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  Ca ²⁺ binds with low affinity to negatively charged residues in disordered regions

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  Ca ²⁺ binding lowers Ddx4 protein net charge, favoring phase separation

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  Ca ²⁺ binding alters condensate stability, dynamics, interfaces, and partitioning

Abstract

Analyzing a family of model biomolecular condensates derived from Ddx4 and Ddx3 proteins over a range of solution conditions, Crabtree et al. show how a balance of repulsive and attractive electrostatic interactions regulate emergent condensate properties, and how these interactions can be tuned by signaling ions such as Ca ²⁺.