Intrinsically disordered linkers determine the interplay between phase separation and gelation in multivalent proteins

Abstract

Phase transitions of linear multivalent proteins control the reversible formation of many intracellular membraneless bodies. Specific non-covalent crosslinks involving domains/motifs lead to system-spanning networks referred to as gels. Gelation transitions can occur with or without phase separation. In gelation driven by phase separation multivalent proteins and their ligands condense into dense droplets, and gels form within droplets. System spanning networks can also form without a condensation or demixing of proteins into droplets. Gelation driven by phase separation requires lower protein concentrations, and seems to be the biologically preferred mechanism for forming membraneless bodies. Here, we use coarse-grained computer simulations and the theory of associative polymers to uncover the physical properties of intrinsically disordered linkers that determine the extent to which gelation of linear multivalent proteins is driven by phase separation. Our findings are relevant for understanding how sequence-encoded information in disordered linkers influences phase transitions of multivalent proteins.

eLife digest

Our cells contain a variety of structures called organelles that perform specific roles within a cell. Some organelles are surrounded by a membrane, while others float inside the cell as spherical droplets made of proteins. These proteins contain several sticky regions, which are connected by flexible linker proteins. It is thought that the level of stickiness and the number of sticky regions, or domains, determine whether a protein will form a membraneless organelle. Often, proteins with similar sticky domains have different linkers, and until now, it was assumed that the linkers do not have any other purpose than stringing the domains together.

To test this further, Harmon et al. used a combination of computer simulations and physics-based theory. In these simulations, the domains were kept the same, but the properties of linkers were changed to see if this would influence how the membraneless organelles are formed.

The results showed that depending on the physical properties of the linkers, the proteins could huddle together and form dense spherical gel-like droplets similar to the membraneless organelles, or form open non-spherical gels. When the linkers were short, the proteins do not easily form droplets. Linkers that were sufficiently long but too bulky, lead to non-spherical gels. Compact linkers, however, enabled proteins to huddle and form spherical gels. The spherical droplet-spanning gels required much less protein compared to the open non-spherical gels.

This suggests that proteins important for forming membraneless organelles can be distinguished from those that are not based on the properties of their linkers – even when their domains are similar.

These findings further scientists’ knowledge of how specific types of proteins form membraneless organelles and will help to understand how membraneless organelles control many key aspects of how a cell works.