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EMBO J. 2017 Oct 16;36(20):2951-2967. doi: 10.15252/embj.201696394. Epub 2017 Aug 8.

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

Author information

1
Department of Pharmacology and Molecular Therapeutics, Uniformed Services University, Bethesda, MD, USA.
2
Neuroscience Graduate Program, Brown University, Providence, RI, USA.
3
Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI, USA.
4
Department of Chemical and Biomolecular Engineering, Lehigh University, Bethlehem, PA, USA.
5
Johns Hopkins Mass Spectrometry and Proteomic Facility, Johns Hopkins University, Baltimore, MD, USA.
6
Graduate Program in Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, RI, USA.
7
Laboratory of Chemical Physics, National Institutes of Health, Bethesda, MD, USA.
8
Department of Pharmacology and Molecular Therapeutics, Uniformed Services University, Bethesda, MD, USA fshewmaker@usuhs.edu nicolas_fawzi@brown.edu.
9
Neuroscience Graduate Program, Brown University, Providence, RI, USA fshewmaker@usuhs.edu nicolas_fawzi@brown.edu.

Abstract

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.

KEYWORDS:

amyotrophic lateral sclerosis; frontotemporal dementia; intrinsically disordered protein; prion; ribonucleoprotein granule

PMID:
28790177
PMCID:
PMC5641905
DOI:
10.15252/embj.201696394
[Indexed for MEDLINE]
Free PMC Article

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