The low-complexity domain of the FUS RNA binding protein self-assembles via the mutually exclusive use of two distinct cross-β cores

Abstract

Significance Single amino acid changes causative of neurologic disease often map to the cross-β forming regions of low-complexity (LC) domains. All such mutations studied to date lead to enhanced avidity of cross-β interactions. The LC domain of the fused in sarcoma (FUS) RNA binding protein contains three different regions that are capable of forming labile cross-β interactions. Here we describe the perplexing effect of amyotrophic lateral sclerosis (ALS)-causing mutations localized to the LC domain of FUS to substantially weaken its ability to form one of its three cross-β interactions. An understanding of how these mutations abet uncontrolled polymerization of the FUS LC domain may represent an important clue as to how LC domains achieve their proper biological function. The low-complexity (LC) domain of the fused in sarcoma (FUS) RNA binding protein self-associates in a manner causing phase separation from an aqueous environment. Incubation of the FUS LC domain under physiologically normal conditions of salt and pH leads to rapid formation of liquid-like droplets that mature into a gel-like state. Both examples of phase separation have enabled reductionist biochemical assays allowing discovery of an N-terminal region of 57 residues that assembles into a labile, cross-β structure. Here we provide evidence of a nonoverlapping, C-terminal region of the FUS LC domain that also forms specific cross-β interactions. We propose that biologic function of the FUS LC domain may operate via the mutually exclusive use of these N- and C-terminal cross-β cores. Neurodegenerative disease–causing mutations in the FUS LC domain are shown to imbalance the two cross-β cores, offering an unanticipated concept of LC domain function and dysfunction.