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

Zachary Monahan, Veronica H. Ryan, Abigail M. Janke, Kathleen A. Burke, Shannon N. Rhoads, Gül H. Zerze, Robert O'Meally, Gregory L. Dignon, Alexander E. Conicella, Wenwei Zheng, Robert B. Best, Robert N. Cole, Jeetain Mittal, Frank Shewmaker, Nicolas L. Fawzi

Research output: Contribution to journalArticle

131 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2951-2967
Number of pages17
JournalEMBO Journal
Volume36
Issue number20
DOIs
StatePublished - Oct 16 2017
Externally publishedYes

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Keywords

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

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Monahan, Z., Ryan, V. H., Janke, A. M., Burke, K. A., Rhoads, S. N., Zerze, G. H., O'Meally, R., Dignon, G. L., Conicella, A. E., Zheng, W., Best, R. B., Cole, R. N., Mittal, J., Shewmaker, F., & Fawzi, N. L. (2017). Phosphorylation of the FUS low-complexity domain disrupts phase separation, aggregation, and toxicity. EMBO Journal, 36(20), 2951-2967. https://doi.org/10.15252/embj.201696394