| 1997 |
TLS/FUS binds RNA in vivo (demonstrated by in vivo UV crosslinking) and undergoes rapid nucleocytoplasmic shuttling (demonstrated by interspecific heterokaryons and cytoplasmic antibody trapping). The RRM domain was found dispensable for in vivo RNA binding, suggesting predominantly non-sequence-specific interactions. Cellular fractionation showed TLS binds RNA in both nucleus and cytoplasm, consistent with an hnRNP-like chaperone function. |
In vivo UV crosslinking, interspecific heterokaryon assay, cytoplasmic antibody injection, cellular fractionation |
Journal of cell science |
High |
9264461
|
| 1994 |
The TLS/FUS protein binds RNA in vitro with preferential binding to poly-G, requiring both amino- and carboxy-terminal RNA-binding motifs. The TLS/FUS N-terminal fusion domain (TFD) in the TLS-ERG chimeric protein functions as a transcriptional activation domain, replacing the ERG N-terminal activation domain, as shown by mutational analysis. |
In vitro RNA binding assay, mutational analysis of fusion protein, transactivation reporter assays |
Oncogene |
Medium |
7970732
|
| 2000 |
TLS/FUS binds RNA polymerase II through its N-terminal domain and recruits serine-arginine (SR) splicing factors through its C-terminal domain. The TLS-ERG leukemia fusion protein retains RNA Pol II binding but loses SR protein recruitment due to replacement of the C-terminus, leading to inhibition of SR-mediated pre-mRNA splicing and altered CD44 splicing in stable K562 cell lines. |
Co-immunoprecipitation, transient transfection splicing assays, stable cell line expression, CD44 splicing analysis |
Molecular and cellular biology |
High |
10779324
|
| 2000 |
Expression of the FUS/TLS-CHOP fusion transgene in mice specifically induces liposarcomas with characteristic lipoblast morphology, intracellular lipid accumulation, induction of adipocyte-specific genes, and high PPARgamma expression, establishing FUS-CHOP overexpression as a key determinant of liposarcoma. |
Transgenic mouse model with EF1alpha-driven FUS-CHOP expression, histological and gene expression analysis |
Oncogene |
High |
10828883
|
| 2000 |
The FUS/TLS domain of FUS-CHOP is required for liposarcoma initiation: transgenic mice expressing CHOP alone (without FUS domain) do not develop tumors, while mice expressing inverted CHOP-FUS (FUS domain fused to C-terminus of CHOP) do develop liposarcomas, establishing a specific and critical role for the FUS domain in transformation. |
Transgenic mouse models comparing FUS-CHOP, CHOP alone, and inverted CHOP-FUS constructs |
Oncogene |
High |
11146553
|
| 2004 |
Structural analysis of human TLS/FUS by limited proteolysis, CD, and NMR revealed that the RRM and zinc finger-like domains form protease-resistant core structures while the RGG repeat regions are unstructured. NMR chemical shift perturbation showed that the zinc finger domain (not the RRM) binds GGUG-containing RNA with Kd ~10^-5 M, suggesting the zinc finger plays a predominant role in RNA recognition. |
Limited proteolysis, MALDI-TOF MS, circular dichroism, NMR spectroscopy (113Cd NMR, amide chemical shift perturbation) |
The Journal of biological chemistry |
High |
15299008
|
| 2008 |
FUS/TLS is phosphorylated at Ser42 in vivo primarily in response to DNA double-strand breaks, with this phosphorylation requiring ATM (not DNA-PK) as established by kinase-specific inhibition and in vitro PIKK phosphorylation assays. Phospho-specific antibodies confirmed in vivo DSB-induced phosphorylation. |
DNA-affinity chromatography, in vitro kinase assay, phospho-specific antibody generation and western blotting, ATM inhibitor experiments |
The Biochemical journal |
High |
18620545
|
| 2009 |
Mutations in FUS/TLS cause its abnormal accumulation in the cytoplasm of neurons (instead of the normal predominantly nuclear localization), identified in familial ALS patients with 13 distinct FUS/TLS mutations. |
Genetic sequencing of familial ALS patients, immunohistochemistry and cellular localization analysis of mutant vs. wild-type FUS in neurons |
Science (New York, N.Y.) |
High |
19251627
|
| 2010 |
TLS/FUS represses RNA polymerase III transcription from all three classes of RNAP III promoters in vitro and associates with RNAP III genes in vivo, possibly via direct interaction with TBP. siRNA depletion of TLS increased RNAP III transcript levels and RNAP III/TBP occupancy at target genes; overexpression decreased RNAP III transcripts. |
In vitro transcription assay, ChIP, siRNA knockdown with RT-qPCR, overexpression studies |
Molecular and cellular biology |
High |
19841068
|
| 2012 |
FUS associates with the SMN complex, mediated by U1 snRNP and by direct protein-protein interactions between FUS and SMN. FUS is required for Gems (Cajal body-related nuclear structures) formation in HeLa cells; ALS-causing FUS mutation R495X also results in Gem loss. Reduction in Gems was observed in ALS patient fibroblasts expressing mutant FUS or TDP-43. |
Co-immunoprecipitation, direct interaction assays, immunofluorescence/Gem counting, patient fibroblast analysis |
Cell reports |
High |
23022481
|
| 2012 |
HITS-CLIP analysis in mouse cerebrum revealed that FUS binding sites tend to form stable secondary structures, that FUS binds scattered sites around alternatively spliced exons (including MAPT, CAMK2A, FMR1), and that FUS binding to promoter antisense strands downregulates coding-strand transcription in a position-dependent manner. |
HITS-CLIP, exon arrays in mouse cortical neurons, bioinformatic analysis |
Scientific reports |
Medium |
22829983
|
| 2013 |
FUS interacts directly with HDAC1, and this interaction is required for FUS recruitment to DNA double-strand break sites and proper DDR signaling. ALS-mutant FUS proteins show diminished interaction with HDAC1 and are defective in DDR and DNA repair. |
Co-immunoprecipitation (FUS-HDAC1), live-cell imaging of FUS recruitment to DSB sites, DNA damage repair assays, patient tissue analysis |
Nature neuroscience |
High |
24036913
|
| 2013 |
RNA binding nucleates the formation of higher-order FUS ribonucleoprotein assemblies that bind the CTD of RNA polymerase II in an RNA-dependent manner, affecting Ser2 phosphorylation and transcription. Both the low-complexity domain and the RGG-rich domain contribute to assembly. The assemblies appear fibrous by electron microscopy with characteristics of β-zipper structures. |
Biochemical assembly assays, electron microscopy, RNA Pol II CTD binding assays, phosphorylation analysis |
Cell reports |
High |
24268778
|
| 2013 |
ALS-linked mutant FUS alters stress granule dynamics: it delays stress granule assembly but once formed, stress granules containing mutant FUS are larger, more dynamic, and more abundant. The RGG domains of FUS are required for its incorporation into stress granules. Arginine methylation within RGG domains does not modulate FUS incorporation into stress granules. |
Live-cell fluorescence imaging, stress granule assembly/disassembly kinetics, domain deletion analysis, methyltransferase inhibitor treatment |
Molecular neurodegeneration |
Medium |
24090136
|
| 2013 |
FUS autoregulates its own protein levels by binding to exon 7 and flanking introns of its own pre-mRNA, repressing exon 7 splicing and promoting nonsense-mediated decay of the exon 7-skipped isoform. ALS mutations causing cytoplasmic FUS mislocalization (R521G, R522G, ΔExon15) show progressively impaired exon 7 repression and autoregulation, correlating with degree of cytoplasmic mislocalization. |
FUS CLIP-seq, splicing reporter assays, siRNA knockdown/rescue, overexpression, antisense oligonucleotides |
PLoS genetics |
High |
24204307
|
| 2013 |
FUS is a prosurvival factor during hyperosmolar stress. Endogenous FUS redistributes from nucleus to cytoplasm and incorporates into stress granules specifically in response to sorbitol (not other stressors like arsenite, H2O2, thapsigargin, or heat shock). This cytoplasmic redistribution is modulated by methyltransferase activity but methyltransferase inhibition does not affect SG incorporation. FUS-depleted cells show reduced viability under hyperosmolar stress. |
Immunofluorescence localization, methyltransferase inhibitors, siRNA knockdown, cell viability assays |
Journal of cellular physiology |
Medium |
23625794
|
| 2014 |
DNA damage (double-strand breaks induced by calicheamicin γ1) causes cytoplasmic translocation of FUS mediated by phosphorylation of its N-terminus by DNA-dependent protein kinase (DNA-PK). This mechanism is distinct from ATM-mediated phosphorylation at Ser42. Cytoplasmic translocation of FUS after DNA damage also involves TAF15, EWS, and Transportin-1. |
Drug treatment (calicheamicin γ1), immunofluorescence, phosphorylation mapping, DNA-PK inhibitor experiments, primary human neurons and astrocytes |
The Journal of neuroscience |
High |
24899704
|
| 2014 |
Nuclear FUS binds active chromatin, and this binding is required for FUS transcription activation (but not alternative splicing regulation). The N-terminal QGSY-rich region (aa 1-164) mediates FUS self-assembly in the mammalian nucleus, which is essential for chromatin binding and transcription activation. RNA binding is also required for FUS self-assembly and chromatin binding. ALS mutations dramatically decrease chromatin binding ability. |
Chromatin fractionation, ChIP, domain deletion mutants, transcription reporter assays, self-assembly assays, RNA-binding mutants |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
25453086
|
| 2014 |
FUS is a co-activator of androgen receptor (AR) in prostate cancer cells. Endogenous FUS co-immunoprecipitates with AR in LNCaP cells, FUS is recruited to the ARE III of the PSA gene enhancer by ChIP, and FUS overexpression enhances while FUS knockdown reduces AR transcriptional activity and androgen-dependent cell proliferation. |
Co-immunoprecipitation of endogenous proteins, ChIP, GAL4 transactivation assay, overexpression and siRNA knockdown |
PloS one |
Medium |
21909421
|
| 2014 |
Activity-dependent regulation of FUS: activation of metabotropic glutamate receptors 1/5 in neocortical slices and synaptoneurosomes increases endogenous mouse FUS and FUS-WT protein levels but decreases FUS-R521G mutant protein, providing a biochemical basis for differential dendritic spine effects between WT and mutant FUS. |
Synaptoneurosomes preparation, mGluR1/5 pharmacological stimulation, western blotting |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
25324524
|
| 2015 |
FUS binds diverse RNA sequences without strong sequence specificity: all five previously proposed binding motifs bind with Kd values spanning only 10-fold, and some RNAs lacking these motifs bind with similar affinity. FUS binds RNA in a length-dependent manner consistent with a substantial non-specific component. FUS binds single-stranded DNA with ~3-fold lower affinity than ssRNA, and double-stranded nucleic acids bind more weakly. |
Quantitative in vitro RNA/DNA binding assays (filter binding, fluorescence polarization), systematic comparison of binding motifs |
Nucleic acids research |
High |
26150427
|
| 2016 |
PINK1 and Parkin are genetic modifiers of FUS-induced neurodegeneration in Drosophila. Downregulating PINK1 or Parkin expression ameliorated FUS-induced neurodegeneration phenotypes. FUS overexpression elevated PINK1 and Parkin protein levels and increased ubiquitinylation of Miro1 (a Parkin E3 ligase substrate). FUS expression reduced mitochondrial axonal transport motility and processivity in motor neurons. |
Drosophila genetic modifier screen, western blotting, mitochondrial transport imaging, ubiquitination assays |
Human molecular genetics |
Medium |
27794540
|
| 2017 |
FUS inclusions lead to mislocalization of specific RNAs from fibroblast cell protrusions and neuronal axons by sequestering kinesin-1 mRNA and protein within inclusions, causing loss of detyrosinated glutamate (Glu)-microtubules and failure of RNA localization. Dissolution of FUS inclusions with engineered Hsp104 disaggregases or kinesin-1 overexpression reverses these effects. Kinesin-1 affects MT detyrosination by targeting tubulin carboxypeptidase enzyme to specific MTs. |
Fluorescence microscopy of RNA localization, kinesin-1 co-recruitment analysis, Hsp104 disaggregase treatment, tubulin modification analysis, rescue experiments |
The Journal of cell biology |
High |
28298410
|
| 2018 |
FUS interacts with the core miRISC component AGO2 and is required for optimal microRNA-mediated gene silencing. FUS promotes silencing by binding to microRNA and mRNA targets (demonstrated for miR-200c and its target ZEB1). The ALS truncation mutant R495X impairs microRNA-mediated gene silencing. The C. elegans homolog fust-1 shares this conserved function. |
Co-immunoprecipitation (FUS-AGO2), RNA binding assays, reporter gene silencing assays, ALS mutant comparison, C. elegans genetic experiments |
Molecular cell |
High |
29499134
|
| 2019 |
Wild-type FUS protein binds to aberrantly retained introns within SFPQ transcripts that are exported from the nucleus into the cytoplasm, providing a mechanism for FUS nuclear-to-cytoplasmic mislocalization in ALS independent of FUS mutations. |
RNA immunoprecipitation, iPSC-derived motor neurons, transgenic mouse models, post-mortem ALS spinal cord analysis |
Brain : a journal of neurology |
Medium |
31368485
|
| 2019 |
Wild-type FUS preferentially binds introns in human motor neuron pre-mRNAs, while ALS mutation (studied in RNA interactome) causes a shift toward 3' UTR binding. ELAVL4 protein levels are increased in ALS-mutant FUS motor neurons; ELAVL4 and mutant FUS interact and co-localize in cytoplasmic speckles with altered biomechanical properties and in stress granules under oxidative stress. |
RNA interactome analysis (iCLIP), immunoprecipitation (FUS-ELAVL4), immunofluorescence, iPSC-derived motor neurons, post-mortem ALS spinal cord |
Cell reports |
Medium |
31242416
|
| 2020 |
FUS acetylation at K510 (in the nuclear localization sequence) disrupts its interaction with Transportin-1, causing cytoplasmic mislocalization and stress granule-like inclusion formation. Acetylation at K315/K316 (in the RNA recognition motif) reduces RNA binding and decreases cytoplasmic inclusion formation. CREB-binding protein/p300 acetylates FUS, while both sirtuins and HDACs contribute to FUS deacetylation. ALS patient fibroblasts show higher FUS K510 acetylation than controls. |
Acetylation site mapping, co-immunoprecipitation (FUS-Transportin-1), site-directed mutagenesis, immunofluorescence, HDAC inhibitor treatment, patient fibroblast analysis |
Human molecular genetics |
High |
32691043
|
| 2020 |
FUS associates with stalled polyribosomes in an mTOR-dependent manner: this association is increased by Torin1 (mTOR kinase inhibitor) or nutrient deprivation but not rapamycin. FUS is required for efficient translational stalling — FUS-deficient cells are refractory to mTOR inhibition-induced translation repression. ALS-linked mutants R521G and P525L associate abundantly with polyribosomes and decrease global protein synthesis; this effect requires RNA-binding by FUS. |
Polyribosome fractionation, Torin1/rapamycin treatment, FUS knockdown/knockout cells, metabolic labeling for translation, RNA-binding mutants |
The Journal of biological chemistry |
Medium |
33082139
|
| 2020 |
FUS ALS-causative mutations induce widespread loss-of-function on splicing, specifically altering intron retention levels in RNA-binding proteins. An intron retention event in FUS itself is associated with its autoregulation and is altered by FUS mutations. This autoregulatory intron retention is also observed in other ALS-linked mutations (TDP-43, VCP, SOD1), suggesting a shared regulatory RNA network. |
High-depth RNA-seq on FUS knockin and knockout models, splicing analysis, comparison across ALS genetic models |
Nucleic acids research |
High |
32479602
|
| 2020 |
EGF receptor (EGFR) phosphorylates FUS at specific tyrosine sites, promoting FUS nuclear translocation. Nuclear FUS binds to the collagen IV promoter, activating transcription. Integrin α1β1 prevents this pathway by inhibiting EGFR. A cell-penetrating peptide that inhibits FUS nuclear translocation reduces collagen IV transcription. |
Tyrosine phosphorylation analysis, EGFR inhibitor treatment, ChIP (FUS at collagen IV promoter), FUS mutagenesis, cell-penetrating peptide, Itga1-null cell comparison |
The Journal of cell biology |
Medium |
32678881
|
| 2021 |
ALS-mutant FUS condenses in axons and sequesters FMRP, promoting FMRP phase separation. This leads to repression of translation in mouse and human FUS-ALS motor neurons. FUS and FMRP co-partition in vitro and repress translation together. Translation of FMRP-bound RNAs is reduced in vivo in FUS-ALS motor neurons. |
Mouse and human iPSC-derived motor neuron models with endogenous knockin mutations, in vitro phase separation assays, translation assays, FMRP interaction studies |
Science advances |
High |
34290090
|
| 2021 |
The FUS gene is dual-coding: an altORF nested in the FUS CDS encodes a conserved 170 amino acid protein (altFUS). altFUS (not FUS itself) is responsible for autophagy inhibition and is pivotal in mitochondrial potential loss and cytoplasmic aggregate accumulation induced by FUS overexpression. Suppression of altFUS in Drosophila protects against FUS-induced neurodegeneration. |
Ribosome profiling, altORF expression constructs, autophagy assays, mitochondrial membrane potential measurements, Drosophila genetics |
EMBO reports |
Medium |
33226175
|
| 2021 |
Crystal structure of the Kapβ2 (Karyopherin-β2/Transportin-1)·FUS(P525L) PY-NLS complex revealed fewer contacts at the mutation site, explaining decreased Kapβ2 affinity. FUS(R495X), despite missing the PY-NLS, binds Kapβ2 via its RGG2 and RGG3 tandem regions competing at the PY-NLS binding site, enabling nuclear localization when arginine methylation is inhibited. Kapβ2 binding also suppresses FUS LLPS. |
Crystal structure determination, biochemical binding assays, cell-based nuclear localization experiments, methylation inhibitor treatment |
Scientific reports |
High |
33580145
|
| 2022 |
Poly(ADP-ribose) (PAR) triggers FUS condensation at 1 nM concentration (1000-fold lower than RNA), through a transient (not stable) interaction with FUS that drives FUS oligomerization. Unlike RNA which stably associates with FUS, PAR interacts transiently. PAR and RNA co-condense with FUS via distinct mechanisms. Inhibition of PARP5a diminishes FUS condensation in cells. |
Single-molecule fluorescence assays, condensate formation assays, PARP5a inhibitor treatment, biochemical binding kinetics |
Molecular cell |
High |
35182479
|
| 2022 |
The FUS::DDIT3 (FUS-CHOP) fusion oncoprotein in myxoid liposarcoma inhibits BAF (mSWI/SNF) complex-mediated chromatin remodeling at adipogenic enhancer sites by sequestering the adipogenic transcription factor CEBPB from the genome, generating a BAF complex loss-of-function phenotype without deleterious BAF subunit mutations. |
ChIP-seq, ATAC-seq, gene expression analysis, BAF ATPase inhibitor treatment, CEBPB co-IP, primary tumor genomics |
Molecular cell |
High |
35390276
|
| 2024 |
Endogenous FUS localizes to mitochondria and interacts with mitochondrial DNA Ligase IIIα (mtLig3), recruiting it to DNA damage sites within mitochondria to maintain mtDNA integrity. ALS-mutant FUS impairs this interaction, hindering mtLig3's repair role and causing increased mtDNA damage and mutations. Targeted introduction of human DNA Ligase 1 restores repair mechanisms and mitochondrial activity in FUS mutant cells. |
Co-immunoprecipitation (FUS-mtLig3), mtDNA damage assays, ALS patient iPSC-derived cells, transgenic mouse model, autopsy samples, DNA Ligase 1 rescue expression |
Nature communications |
High |
38461154
|
| 2024 |
Reduction of m6A RNA modification (via METTL3 inhibitor STM-2457) decreases the number and accelerates dissolution of FUS-containing stress granules/cytoplasmic inclusions in neuronal cells, iPSC-derived motor neurons, and patient-derived fibroblasts expressing mutant FUS. Cells expressing mutant FUS show higher m6A levels, suggesting m6A homeostasis influences pathological FUS aggregate formation. |
METTL3 inhibitor treatment (STM-2457), stress granule imaging, m6A quantification, iPSC-derived motor neurons, patient fibroblasts, transcriptome analysis |
Nature communications |
Medium |
38866783
|