The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization

Rhoads, Shannon N; Monahan, Zachary; Yee, Debra; Leung, A.Y.T.; Newcombe, Cameron G.; O’Meally, Robert N.; Cole, Robert N.; Shewmaker, Frank

doi:10.1091/mbc.e17-12-0735

Cited by 44 publications

(88 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One example is the neurodegeneration-linked protein FUS, which is phosphorylated at multiple Ser/Thr residues by DNAdependent protein kinase (DNA-PK) (75)(76)(77). Phosphorylation with DNA-PK interferes with phase separation of the FUS LCD (30, 76, 78), reduces binding to FUS-LCD hydrogels (79), and abrogates phase separation of multivalent interacting proteins JBC REVIEWS: Control of phase separation and RNP granule dynamics by PTMs tethered to FUS-LCD (80).…”

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

“…Under which conditions FUS is phosphorylated in vivo is still poorly understood; so far, only the treatment of cells with DNA damage-inducing drugs, e.g. calicheamicin or calyculin-A, was found to transiently induce FUS phosphorylation on multiple Ser/Thr residues (75,77). It has been proposed that transient phosphorylation of FUS after DNA damage may occur in FTD patients with FUS aggregates, as post-mortem brains of these patients show elevated levels of ␥-H2AX (75), a marker for DNA double-strand breaks.…”

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

“…It has been proposed that transient phosphorylation of FUS after DNA damage may occur in FTD patients with FUS aggregates, as post-mortem brains of these patients show elevated levels of ␥-H2AX (75), a marker for DNA double-strand breaks. Whether phosphorylation of FUS (and the other members of the FET protein family, EWS and TAF15) indeed occurs in disease conditions or as a physiological stress response, and how it affects other functional properties, such as protein interactions or subcellular localization, are still matters of debate (22,75,77) and remain to be clarified by future studies.…”

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

See 2 more Smart Citations

Friend or foe—Post-translational modifications as regulators of phase separation and RNP granule dynamics

Hofweber¹,

Dormann

2019

Journal of Biological Chemistry

306

294

View full text Add to dashboard Cite

Edited by Paul E. FraserRibonucleoprotein (RNP) granules are membrane-less organelles consisting of RNA-binding proteins (RBPs) and RNA. RNA granules form through liquid-liquid phase separation (LLPS), whereby weak promiscuous interactions among RBPs and/or RNAs create a dense network of interacting macromolecules and drive the phase separation. Post-translational modifications (PTMs) of RBPs have emerged as important regulators of LLPS and RNP granule dynamics, as they can directly weaken or enhance the multivalent interactions between phase-separating macromolecules or can recruit or exclude certain macromolecules into or from condensates. Here, we review recent insights into how PTMs regulate phase separation and RNP granule dynamics, in particular arginine (Arg)-methylation and phosphorylation. We discuss how these PTMs regulate the phase behavior of prototypical RBPs and how, as "friend or foe," they might influence the assembly, disassembly, or material properties of cellular RNP granules, such as stress granules or amyloidlike condensates. We particularly highlight how PTMs control the phase separation and aggregation behavior of disease-linked RBPs. We also review how disruptions of PTMs might be involved in aberrant phase transitions and the formation of amyloid-like protein aggregates as observed in neurodegenerative diseases.

show abstract

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

Section: Phosphorylation Regulates Llps and Aggregation Of Disease-limentioning

confidence: 99%

See 1 more Smart Citation

Friend or foe—Post-translational modifications as regulators of phase separation and RNP granule dynamics

Hofweber¹,

Dormann

2019

Journal of Biological Chemistry

306

294

View full text Add to dashboard Cite

show abstract

“…Using solid-state NMR, Murray and coworkers identified a core region of FUS’s PrLD (amino acids 39–95) that forms amyloid with parallel in-register β sheet structure [ 35 ]. Importantly, this region overlaps with multiple putative sites of phosphorylation ( Table 1 ) [ 33 ]; this is predicted to have strong influence on FUS’s capacity to form solid aggregates (discussed below).…”

Section: Fus Structure and Functionmentioning

confidence: 99%

The Role of Post-Translational Modifications on Prion-Like Aggregation and Liquid-Phase Separation of FUS

Rhoads

Monahan

Yee

et al. 2018

IJMS

101

View full text Add to dashboard Cite

Subcellular mislocalization and aggregation of the human FUS protein occurs in neurons of patients with subtypes of amyotrophic lateral sclerosis and frontotemporal dementia. FUS is one of several RNA-binding proteins that can functionally self-associate into distinct liquid-phase droplet structures. It is postulated that aberrant interactions within the dense phase-separated state can potentiate FUS's transition into solid prion-like aggregates that cause disease. FUS is post-translationally modified at numerous positions, which affect both its localization and aggregation propensity. These modifications may influence FUS-linked pathology and serve as therapeutic targets. Keywords: FUS; ALS; FTLD; prion; amyloid; LLPS The Link between FUS and Neurodegenerative DiseaseFUS (fused in sarcoma) gets its name from forming oncogenic fusion proteins with specific transcription factors following chromosomal rearrangements [1]. Such rearrangement is common in liposarcomas, thus FUS also goes by the name TLS (translocated in liposarcoma). Since 2009, FUS has received more attention for its connection to neurodegenerative disease after missense mutations were discovered to cause a small percentage of cases of amyotrophic lateral sclerosis (ALS-FUS) [2,3]. In the motor neurons of these patients, the normally nuclear FUS protein was found in cytoplasmic proteinaceous inclusions. Since then, non-mutant FUS has been identified in cytoplasmic inclusions in cortical neurons of a subset of patients with frontotemporal dementia (FTD; the neuropathological diagnosis is termed frontotemporal lobar degeneration (FTLD-FUS)) [4]. Both ALS and FTD are incurable and their clinical and pathological overlap suggests that they are part of a disease continuum.Mutations in FUS are autosomal dominant causes of familial ALS. Most mutations alter the C-terminal nuclear localization signal, resulting in excess cytoplasmic FUS that can form inclusions with gain-of-function toxicity [5]. Whether ALS-FUS or FTLD-FUS, it is the accumulation of FUS into cytoplasmic aggregates that appears to cause neuronal loss. The clinical presentation likely depends on the specific type of neurons that are affected. Biophysical and histological analysis suggest that FUS cytoplasmic aggregation may spread across anatomical networks through a prion-like mechanism [6]. Therefore, future drugs may target FUS's ability to cytoplasmically localize and/or form proteinaceous aggregates. Extensive post-translational methylation and phosphorylation of FUS have been shown to influence localization and aggregation, respectively. Here we review the post-translational modifications (PTMs) of FUS in the context of how they affect function, self-association and pathology.

show abstract

“…Recently, PARG activity has been also associated with an increased presence of FUS in the cytoplasm of neurons after stress (Naumann et al, 2018). However, phosphorylation of FUS residues in the LCD, notably by DNA-dependent protein kinase (DNA-PK) (Deng et al, 2014;Monahan et al, 2017), and methylation of the RGG domains (Kaneb et al, 2012) may also participate in the nucleocytoplasmic shuttling, even if this is still a debated issue (Rhoads et al, 2018). The PARGdependent translocation of FUS provides an interesting link between DNA repair and neurodegenerative diseases (Naumann et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

PARP-1 Activation Directs FUS to DNA Damage Sites to Form PARG-Reversible Compartments Enriched in Damaged DNA

Singatulina¹,

Hamon²,

Sukhanova³

et al. 2019

Cell Reports

167

181

View full text Add to dashboard Cite

Graphical Abstract Highlights d FUS is directed to DNA damage sites by binding PAR synthesized by PARP-1 d FUS then forms large compartments in which damaged DNA is concentrated d PARG dissociates damaged DNA compartments by hydrolyzing PAR d Then FUS shuttles from the nucleus to the cytoplasm Correspondence lavrik@niboch.nsc.ru (O.I.L.), david.pastre@univ-evry.fr (D.P.) In Brief Using a single-molecule approach, Singatulina et al. reconstitute the DNA repair system leading to the specific recruitment of FUS, an mRNA-binding protein related to liquid-liquid phase separation biology, to DNA damage sites, thus revealing the capacity of FUS to form dynamic compartments in which damaged DNA is concentrated. SUMMARY PARP-1 synthesizes long poly(ADP-ribose) chains (PAR) at DNA damage sites to recruit DNA repair factors. Among proteins relocated on damaged DNA, the RNA-binding protein FUS is one of the most abundant, raising the issue about its involvement in DNA repair. Here, we reconstituted the PARP-1/ PAR/DNA system in vitro and analyzed at the single-molecule level the role of FUS. We demonstrate successively the dissociation of FUS from mRNA, its recruitment at DNA damage sites through its binding to PAR, and the assembly of damaged DNA-rich compartments. PARG, an enzyme family that hydrolyzes PAR, is sufficient to dissociate damaged DNArich compartments in vitro and initiates the nucleocytoplasmic shuttling of FUS in cells. We anticipate that, consistent with previous models, FUS facilitates DNA repair through the transient compartmentalization of DNA damage sites. The nucleocytoplasmic shuttling of FUS after the PARG-mediated compartment dissociation may participate in the formation of cytoplasmic FUS aggregates.

show abstract

The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization

Cited by 44 publications

References 56 publications

Friend or foe—Post-translational modifications as regulators of phase separation and RNP granule dynamics

Friend or foe—Post-translational modifications as regulators of phase separation and RNP granule dynamics

The Role of Post-Translational Modifications on Prion-Like Aggregation and Liquid-Phase Separation of FUS

PARP-1 Activation Directs FUS to DNA Damage Sites to Form PARG-Reversible Compartments Enriched in Damaged DNA

Contact Info

Product

Resources

About