Mutant FUS causes DNA ligation defects to inhibit oxidative damage repair in Amyotrophic Lateral Sclerosis

Weng, Haibo; Guo, Wenting; Mitra, Joy; Hegde, Pavana M.; Vandoorne, Tijs; Eckelmann, Bradley; Mitra, Sankar; Tomkinson, Alan E.; Bosch, Ludo Van Den; Hegde, Muralidhar L.

doi:10.1038/s41467-018-06111-6

Cited by 152 publications

(190 citation statements)

References 68 publications

Supporting

Mentioning

185

Contrasting

Order By: Relevance

“…In light of the results presented here, we may also ask whether pathogenic FUS mutations, notably in the RGG repeats, impair DNA repair in damaged DNA-rich compartments and/or the nucleocytoplasmic shuttling of FUS. These issues deserve to be addressed to increase our understanding of the link between DNA repair and neurodegenerative diseases (Hoch et al, 2017;Naumann et al, 2018;Wang et al, 2018a).…”

Section: Discussionmentioning

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

154

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

Section: Discussionmentioning

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

154

181

View full text Add to dashboard Cite

show abstract

“…FUS DNA repair functions have been deduced from its PARP-dependent recruitment to sites of microirradiation; its coimmunoprecipitation with repair proteins; and the modest chromosome instability and DSB repair defects of FUS-deficient cells (Deng et al, 2014;Hicks et al, 2000;Martinez-Macias et al, 2019;Mastrocola et al, 2013;Rulten et al, 2014;Singatulina et al, 2019;Wang et al, 2018;Wang et al, 2013). Despite these results, a unifying role for FUS in genome proteciotn has yet to emerge.…”

Section: Discussionmentioning

confidence: 99%

Fused in sarcoma regulates DNA replication timing and progression

Jia

Scalf

Tonzi

et al. 2020

Preprint

View full text Add to dashboard Cite

Fused in sarcoma (FUS) encodes a low complexity RNA-binding protein with diverse roles in transcriptional activation and RNA processing.While oncogenic fusions of FUS and transcription factor DNA-binding domains are associated with soft tissue sarcomas, dominant mutations in FUS cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). FUS has also been implicated in DNA double-strand break repair (DSBR) and genome maintenance. However, the underlying mechanisms are unknown. Here we employed quantitative proteomics, trancriptomics, and DNA copy number analysis (Sort-Seq), in conjunction with FUS -/cells to ascertain roles of FUS in genome protection. FUS-deficient cells exhibited alterations in the recruitment and retention of DSBR factors BRCA1 and 53BP1 but were not overtly sensitive to genotoxins. FUS-deficient cells exhibited reduced proliferative potential that correlated with reduced replication fork speed, diminished loading of prereplication complexes, and attenuated expression of S-phase associated genes. FUS interacted with replisome components, including lagging strand synthesis factors, but did not translocate with active replication forks. Finally, we show that FUS contributes to genome-wide control of DNA replication timing. Alterations in DNA replication initiation and timing may contribute to genome instability and functional defects in mitotically active cells harboring disease-associated FUS fusions FUS regulates pre-replication complex (pre-RC) loading and associates with DNA replication factors. Given their reduced DNA replication rate, we investigated whether FUS -/cells exhibited defects in the chromatin loading of replication licensing factors, including the origin recognition complex (ORC), CDC6, and CDT,and the MCM replicative helicase (Fragkos et al., 2015)). Mitotically arrested FUS +/+ , FUS -/-, and FUS -/-:FUS cells were released into early G 1 phase and soluble and chromatin fractions analyzed by immunoblotting. FUS-deficient cells showed normal cell progression from G 2 /M to G 1 phase and unchanged ORC loading onto chrmatin in G 1 (Fig. 5A and B). By contrast, recruitment of CDC6 and CDT1 was significantly decreased in FUS -/cells and rescued by FUS reexpression (Fig. 5B). As expected, CDC6 and CDT1-dependent loading of MCM complex was also reduced in FUS -/cells (Sup. Fig. 6). Collectively, these results revealed that FUS facilitates ORC-dependent recruitment of pre-RC factors CDC6 and CDT1 to replication origins.

show abstract

“…Preventing the assembly of DNA repair foci via PARP1 inhibition leads to neurodegenerative phenotypes, such as decreased organelle transport, in ALS‐patient‐derived motor neurons . Moreover, ALS‐associated mutations of a DNA repair foci component FUS decreases its recruitment to DNA damage sites, reducing the recruitment of other DNA repair proteins, such as the damage‐sensing kinase ATM and the double‐strand break recognition factor NBS1 . How defects in DNA repair lead to neurodegeneration remains an open question, but a prolonged stress response triggered by unresolved DNA damage and subsequent cellular senescence likely exacerbates disease phenotypes …”

Section: Biomolecular Condensates In Neurodegenerationmentioning

confidence: 99%

“…The deletion of FUS and EWS has been linked to increased genome instability and radiation sensitivity in mice . In cultured human cells, FUS depletion leads to sensitivity to single‐ and double‐strand breaks and oxidative damage, as well as reduced efficiency of homologous recombination and nonhomologous end joining . Based on existing knowledge of FET protein functions in DNA repair, the early DNA repair foci may “hold” broken DNA together (pairing of homologous DNA strands in vitro), facilitate DNA ligation (interacting with DNA ligase III‐XRCC1 complex and enhancing their in vitro ligase activity) or augment DNA damage signaling (interacting with HDAC1) (Figure C).…”

Section: Biomolecular Condensates In Cancermentioning

confidence: 99%

Biomolecular condensates in neurodegeneration and cancer

Spannl

Tereshchenko

Mastromarco

et al. 2019

Traffic

View full text Add to dashboard Cite

The intracellular environment is partitioned into functionally distinct compartments containing specific sets of molecules and reactions. Biomolecular condensates, also referred to as membrane‐less organelles, are diverse and abundant cellular compartments that lack membranous enclosures. Molecules assemble into condensates by phase separation; multivalent weak interactions drive molecules to separate from their surroundings and concentrate in discrete locations. Biomolecular condensates exist in all eukaryotes and in some prokaryotes, and participate in various essential house‐keeping, stress‐response and cell type‐specific processes. An increasing number of recent studies link abnormal condensate formation, composition and material properties to a number of disease states. In this review, we discuss current knowledge and models describing the regulation of condensates and how they become dysregulated in neurodegeneration and cancer. Further research on the regulation of biomolecular phase separation will help us to better understand their role in cell physiology and disease.

show abstract

Mutant FUS causes DNA ligation defects to inhibit oxidative damage repair in Amyotrophic Lateral Sclerosis

Cited by 152 publications

References 68 publications

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

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

Fused in sarcoma regulates DNA replication timing and progression

Biomolecular condensates in neurodegeneration and cancer

Contact Info

Product

Resources

About