Phosphorylation in the Prion-Like Domain of Fused-in-Sarcoma: A Critical Mediator of Aggregation and Toxicity
Abstract
Neuronal cytoplasmic inclusions of aggregated RNA-binding protein fused in sarcoma (FUS) are hallmarks of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) subtypes. Intriguingly, FUS's nearly uncharged, aggregation-prone, yeast prion-like, low sequence-complexity domain (PrLD, also LC) is known to be targeted for phosphorylation. Here we map in vitro and in-cell phosphorylation sites across FUSPrLD. We show that both phosphorylation and phosphomimetic variants reduce its aggregation-prone/prion-like character, disrupting FUS phase separation in the presence of RNA or salt and reducing FUS propensity to aggregate. Nuclear magnetic resonance spectroscopy demonstrates the intrinsically disordered structure of FUS PrLD is preserved after phosphorylation; however, transient domain collapse and self-interaction are reduced by phosphomimetics. Moreover, we show that phosphomimetic FUS reduces aggregation in human and yeast cell models, and can ameliorate FUS-associated cytotoxicity. Hence, post-translational modification may be a mechanism by which cells control physiological assembly and prevent pathological protein aggregation, suggesting a potential treatment pathway amenable to pharmacologic modulation.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 13, 2018
- Accession Number
- AD1128512
Entities
People
- Zachary Monahan
Organizations
- Uniformed Services University of the Health Sciences