ATPase-Modulated Stress Granules Contain a Diverse Proteome and Substructure

Jain, Shuchi; Wheeler, Joshua; Walters, Robert W.; Agrawal, Anurag; Barsic, Anthony; Parker, Roy

doi:10.1016/j.cell.2015.12.038

Cited by 1,269 publications

(1,827 citation statements)

References 55 publications

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“…Further efforts are needed to determine how phase separation is governed in cells where a multitude of components make up physiological granules. FUS‐containing membraneless organelles are not homogeneous structures composed of one protein and one RNA (Kato et al , 2012; Jain et al , 2016). The addition of other granule components and molecular chaperones in vivo may decrease the dominant role of charge–charge repulsion in the effect on phase separation observed in vitro .…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2017

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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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2017

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“…Similar to yeast cells, liquid-like assemblies in mammalian cells are dissolved through hexanediol Similar to yeast P bodies, mammalian stress granules dissolve in a hexanedioldependent manner over a time course of 10 to 20 min [2]. This is due to the fact that stress granules in mammalian cells are much more liquid-like than stress granules in yeast cells [2] [14]. To further investigate this, we tested the concentration dependence of the hexanediol effect.…”

Section: Liquid-like But Not Solid-like Assemblies In Living Yeast Cementioning

confidence: 99%

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Kroschwald

Maharana

Alberti³

2017

Matters

277

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Membrane-less compartments, such as P bodies and stress granules, play major roles in cell biology and disease. The dynamics and physical properties of these compartments are very diverse, ranging from liquid-like to solid-like behavior. Importantly, changes in compartment dynamics have been linked to changes in compartment function or pathology. However, appropriate tools to investigate the physical properties of membrane-less compartments are still lacking. The aliphatic alcohol hexanediol has been proposed as a tool to differentiate between liquid-like and solid-like assemblies in living cells. We show that in vitro reconstituted compartments formed by the RNAbinding protein FUS are rapidly dissolved by hexanediol. In contrast, solid-like fibers of FUS, which have been linked to the disease amyotrophic lateral sclerosis (ALS), are resistant to hexanediol treatment, supporting the idea that hexanediol can differentiate between liquid-and solid-like assemblies. We further show that hexanediol can be used to examine the physical properties of membrane-less compartments in vivo. We find that hexanediol dissolves dynamic, liquid-like assemblies, such as P bodies, whereas solid-like assemblies, such as protein aggregates and cytoskeletal assemblies, are largely resistant to hexanediol. Finally, we report here that extended exposure of yeast and mammalian cells to hexanediol is cytotoxic and causes abnormal changes in cell morphology, which trigger the formation of aberrant assemblies. We therefore urge care in the use of hexanediol, especially when cells are exposed to hexanediol for extended times. In summary, we propose that hexanediol is a powerful tool to probe the physical properties of membrane-less compartments, but only if adequate controls are included and precautions are taken.

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“…In yeast and humans, SGs contain a diverse proteome (Jain et al, 2016). Some RBPs are essential for SG formation in these species, such as T-cell-restricted intracellular antigen-1 (TIA-1) and TIA-1-related (TIAR) proteins (Gilks et al, 2004), Ras-GTPase-activating protein SH3-domain-binding proteins (G3BPs; Aulas et al, 2015), and Tudor staphylococcal nucleases (TSNs; Gao et al, 2015).…”

Section: Mrna Sequestration In Sgs: a Transit Depot Between Translatimentioning

confidence: 99%

Polysomes, Stress Granules, and Processing Bodies: A Dynamic Triumvirate Controlling Cytoplasmic mRNA Fate and Function

2017

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ATPase-Modulated Stress Granules Contain a Diverse Proteome and Substructure

Cited by 1,269 publications

References 55 publications

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

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

Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments

Polysomes, Stress Granules, and Processing Bodies: A Dynamic Triumvirate Controlling Cytoplasmic mRNA Fate and Function

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