Motif-pattern dependence of biomolecular phase separation driven by specific interactions

Eukaryotic cells partition a wide variety of important materials and processes into biomolecular condensates—phase-separated droplets that lack a membrane. In addition to nonspecific electrostatic or hydrophobic interactions, phase separation also depends on specific binding motifs that link together constituent molecules. Nevertheless, few rules have been established for how these ubiquitous specific, saturating, motif-motif interactions drive phase separation. By integrating Monte Carlo simulations of lattice-polymers with mean-field theory, we show that the sequence of heterotypic binding motifs strongly affects a polymer’s ability to phase separate, influencing both phase boundaries and condensate properties (e.g. viscosity and polymer diffusion). We find that sequences with large blocks of single motifs typically form more inter-polymer bonds, which promotes phase separation. Notably, the sequence of binding motifs influences phase separation primarily by determining the conformational entropy of self-bonding by single polymers. This contrasts with systems where the molecular architecture primarily affects the energy of the dense phase, providing a new entropy-based mechanism for the biological control of phase separation.Author summary: Cells need to concentrate biomolecules in the right place at the right time in order to function. Many important intracellular compartments are liquid droplets formed by phase separation, the same process that separates oil from vinegar. The properties of such “biomolecular condensates” depend on the component molecules, such as proteins and RNAs. These molecules are polymers made of many interacting monomers, often organized into “motifs,” and the sequence of motifs shapes the properties of the condensates. Recent work has revealed important principles governing phase separation when the motifs are charged and interact across long distances, but many phase-separating molecules form specific interactions that are short-range and one-to-one. How does the sequence of specifically-interacting motifs affect phase separation? Using a combination of simulations and theoretical calculations, we show that the sequence has profound effects on both the formation and properties of condensates. Sequences with large blocks of identical motifs are better at phase separating but more viscous and solid-like. Importantly, we find that sequence controls phase separation via the proclivity to form self-bonds instead of forming bonds with other polymers. Thus the sequence of specifically-interacting motifs provides a control point for the formation and properties of phase-separated intracellular compartments.