Phosphorylation of the FUS low‐complexity domain disrupts phase separation, aggregation, and toxicity

• Zachary Monahan ^[4] ; Veronica H Ryan ^[1] ; Abigail M Janke ^[1] ; Kathleen A Burke ^[1] ; Shannon N Rhoads ^[4] ; Gül H Zerze ^[2] ; Robert O'Meally ^[3] ; Gregory L Dignon ^[2] ; Alexander E Conicella ^[1] ; Wenwei Zheng ^[5] ; Robert B Best ^[5] ; Robert N Cole ^[3] ; Jeetain Mittal ^[2] ; Frank Shewmaker ^[4] ; Nicolas L Fawzi ^[1]
  1. [1] Brown University

     Brown University

     City of Providence, Estados Unidos

     
  2. [2] Lehigh University

     Lehigh University

     City of Bethlehem, Estados Unidos

     
  3. [3] Johns Hopkins University

     Johns Hopkins University

     Estados Unidos

     
  4. [4] 1 Department of Pharmacology and Molecular Therapeutics Uniformed Services University Bethesda MD USA
  5. [5] 7 Laboratory of Chemical Physics National Institutes of Health Bethesda MD USA

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• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 36, Nº. 20, 2017, págs. 2951-2967
• Idioma: inglés
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  • Neuronal inclusions of aggregated RNA‐binding protein fused in sarcoma (FUS) are hallmarks of ALS and frontotemporal dementia subtypes. Intriguingly, FUS's nearly uncharged, aggregation‐prone, yeast prion‐like, low sequence‐complexity domain (LC) is known to be targeted for phosphorylation. Here we map in vitro and in‐cell phosphorylation sites across FUS LC. 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 LC 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.