Affinage

EWSR1

RNA-binding protein EWS · UniProt Q01844

Length
656 aa
Mass
68.5 kDa
Annotated
2026-06-09
100 papers in source corpus 33 papers cited in narrative 33 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 5/6 claims corpus-supported (83%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

EWSR1 is a multifunctional RNA/DNA-binding protein whose N-terminal low-complexity, prion-like domain undergoes tyrosine-driven phase separation and mediates homo- and heterotypic protein assemblies, while its C-terminal RNA-binding region directs RNA-dependent self-association (PMID:14534527, PMID:38492239). In its wild-type role, EWSR1 supports genome integrity and mitotic fidelity: it suppresses DNA damage-induced R-loops to promote BRCA1-mediated homologous recombination (PMID:29513652), maintains centromere identity by binding CENP-A through its SYGQ2 prion-like region and centromeric R-loops via its RNA-recognition motif (PMID:37243594), and recruits Aurora B to the anaphase midzone through the R565 residue of its RGG3 domain to enable faithful division (PMID:25483190), consistent with the spindle and apoptotic defects seen upon its loss (PMID:17912356). EWSR1 acts in transcription and pre-mRNA splicing, cooperating with REST to repress neuronal genes (PMID:24069508) and controls miRNA biogenesis by modulating Drosha to govern target genes during development (PMID:24185621). It also sustains mitochondrial homeostasis by protecting PGC-1α from FBXW7-dependent proteasomal degradation (PMID:25918410), interacts with lamin A/C, and is required in vivo for B-cell development, meiosis, and suppression of premature senescence (PMID:17415412, PMID:21030557). The protein is heavily asymmetrically dimethylated on arginines within its RGG motifs, a modification controlling its abundance, nuclear/cell-surface localization (PMID:11278906, PMID:12915128). EWSR1 is recurrently the 5' partner in oncogenic fusions (EWS-FLI1, EWS-ATF1, EWSR1-NR4A3, EWS-WT1) where its intrinsically disordered N-terminal domain functions as a phase-separating transactivation module appended to a heterologous DNA-binding domain, reprogramming chromatin and 3D genome architecture and corrupting splicing (PMID:15491164, PMID:25453903, PMID:12923058, PMID:34035145, PMID:35477713).

Mechanistic history

Synthesis pass · year-by-year structured walk · 23 steps
  1. 1994 Medium

    Established that in the EWS-FLI1 fusion the heterologous ETS domain, not the EWS portion, dictates sequence-specific DNA binding, localizing the contribution of EWS to transactivation rather than recognition.

    Evidence in vitro binding-site selection and truncation DNA-binding assays with epitope-tagged protein

    PMID:7517940

    Open questions at the time
    • Did not address the function of wild-type EWSR1
    • Did not test fusion activity in chromatin context
  2. 1993 Medium

    Defined the EWS N-terminal domain as a transcriptional regulatory module that modulates the activity of the fused FLI1 domain, framing the fusion as a chimeric transcription factor.

    Evidence transcriptional reporter assays with deletion constructs in transfected cells

    PMID:7503813

    Open questions at the time
    • Mechanism of activation not biochemically defined
    • Wild-type EWSR1 transcriptional role not addressed
  3. 2001 Medium

    Connected EWSR1 to the splicing machinery and showed the fusion perturbs splice-site selection, opening a non-transcriptional axis of EWSR1 function.

    Evidence in vivo E1A splicing assay, mutational analysis and interaction assays with SF1/U1C

    PMID:11301318

    Open questions at the time
    • Direct RNA targets in vivo not mapped
    • Splicing role of wild-type EWSR1 incompletely resolved
  4. 2001 High

    Mapped extensive asymmetric arginine dimethylation across the RGG motifs and detected EWS at the cell surface, establishing a post-translational code and unexpected localization.

    Evidence cell-surface biotinylation, isoelectric focusing and tandem mass spectrometry site mapping

    PMID:11278906

    Open questions at the time
    • Responsible methyltransferase not identified
    • Functional consequence of each methylation site untested
  5. 2003 High

    Showed EWSR1 self-associates in an RNA-dependent manner via its C-terminal RNA-binding domain while the N-terminal domain mediates homo/heterotypic interactions, defining the basis for its higher-order assembly.

    Evidence FRET, mammalian two-hybrid, GST pull-down, Co-IP with RNaseA sensitivity

    PMID:14534527

    Open questions at the time
    • Stoichiometry of assemblies not determined
    • Link to phase separation not yet drawn
  6. 2003 Medium

    Demonstrated that arginine methylation controls EWS abundance and partitioning between nucleus and cell surface, with mitogenic signals tuning surface exposure.

    Evidence methylation-inhibitor treatment, cell-surface biotinylation, mitogenic stimulation of T cells

    PMID:12915128

    Open questions at the time
    • Pharmacological inhibition is not site-specific
    • Mechanism linking methylation to localization unresolved
  7. 2004 Medium

    Established that the fusion protein is intrinsically disordered yet retains DNA binding and transcriptional activity, anchoring the concept that EWSR1's disordered domain is functionally active without folding.

    Evidence recombinant protein purification, circular dichroism, fluorescence spectroscopy, in vitro DNA-binding/transcription assays

    PMID:15491164

    Open questions at the time
    • Behavior of full-length protein in cells not addressed
    • Condensate behavior not yet examined
  8. 2006 High

    Identified RNA helicase A as a direct cofactor that co-occupies fusion target promoters and amplifies oncogenic transcription and transformation.

    Evidence phage display, GST pull-down, ELISA, reciprocal Co-IP, ChIP, reporter and anchorage-independent growth assays

    PMID:16740692

    Open questions at the time
    • Whether wild-type EWSR1 uses the same interaction surface untested
    • Genome-wide co-occupancy not mapped
  9. 2007 High

    Defined essential in vivo roles for EWSR1 in B-cell development, meiosis, and senescence suppression and identified lamin A/C as a partner, establishing physiological functions beyond cancer.

    Evidence Ews knockout mice, lymphocyte/meiosis analysis, MEF senescence and irradiation assays, Co-IP

    PMID:17415412

    Open questions at the time
    • Molecular basis of meiotic and DNA-damage phenotypes not resolved
    • Direct vs indirect lamin A/C effects unclear
  10. 2007 Medium

    Showed EWSR1 is required for mitotic spindle integrity across zebrafish and human cells, linking its loss to chromosomal instability and apoptosis.

    Evidence morpholino knockdown in zebrafish, siRNA in HeLa, mitotic spindle imaging

    PMID:17912356

    Open questions at the time
    • Molecular mechanism at the spindle not defined here
    • Knockdown specificity not exhaustively controlled
  11. 2010 Medium

    Established EWSR1 as a regulator of hematopoietic stem cell quiescence whose loss triggers p16-driven senescence.

    Evidence Ews knockout mice, HSPC flow cytometry, β-galactosidase and p16INK4a assays

    PMID:21030557

    Open questions at the time
    • Direct molecular targets in HSCs not identified
    • Relationship to senescence pathways in other tissues unclear
  12. 2013 Medium

    Showed EWSR1 controls miRNA biogenesis by restraining Drosha, defining a post-transcriptional output that shapes developmental gene expression.

    Evidence Ews-null MEFs, miRNA qPCR, Drosha knockdown rescue, target immunoblotting

    PMID:24185621

    Open questions at the time
    • Mechanism of Drosha regulation not resolved
    • Direct vs indirect control of Drosha expression unclear
  13. 2013 Medium

    Identified a wild-type EWSR1–REST complex on chromatin that cooperatively silences neuronal genes and restrains transformation.

    Evidence Co-IP, ChIP-seq, RNA-seq after RNAi, transformation assays

    PMID:24069508

    Open questions at the time
    • Direct DNA-binding role of EWSR1 within the complex unclear
    • Single-lab ChIP-seq, no reciprocal validation
  14. 2014 High

    Resolved two divergent gene-regulatory mechanisms of EWS-FLI1 — de novo enhancer creation at GGAA microsatellites and inactivation of native ETS enhancers — explaining its dual activation/repression output.

    Evidence ChIP-seq, ATAC/DNaseI-seq, chromatin conformation capture, knockdown/rescue, reporter assays

    PMID:25453903

    Open questions at the time
    • Role of EWS phase separation in enhancer formation not addressed here
    • Wild-type EWSR1 enhancer behavior not compared
  15. 2014 High

    Demonstrated that EWSR1 fusions to other DNA-binding partners (WT1, ATF1) generate novel binding specificities and target programs, generalizing the chimeric-transactivator model across sarcomas.

    Evidence in vitro/in vivo binding assays, ChIP, mutagenesis, transgenic and inducible expression models

    PMID:12923058 PMID:23281395 PMID:24934812

    Open questions at the time
    • How the EWS domain alters partner specificity mechanistically not resolved
    • Direct target sets incompletely catalogued
  16. 2014 Medium

    Mapped EWSR1 recruitment of Aurora B to the midzone via the RGG3 R565 residue, providing a molecular basis for its role in cytokinesis and ploidy control.

    Evidence siRNA knockdown, midzone imaging, Co-IP, domain/point mutation and rescue

    PMID:25483190

    Open questions at the time
    • Whether methylation of R565 regulates the interaction untested
    • Reciprocal validation of Aurora B interaction limited
  17. 2015 High

    Defined EWS-FLI1 as a splicing regulatory hub binding nascent RNA and spliceosomal factors, and showed RHA helicase inhibition by the fusion, with both reversible by YK-4-279.

    Evidence CLIP-seq, exon array/RNA-seq, Co-IP with DDX5/hnRNP K/PRPF6, in vitro helicase/ATPase assays, enantiomer-specific inhibition

    PMID:25564528 PMID:25737553

    Open questions at the time
    • Splicing role of wild-type EWSR1 not directly compared
    • In vivo significance of altered isoforms incompletely tested
  18. 2018 High

    Established that EWSR1 suppresses R-loops to enable homologous recombination, and that the fusion's transcriptional excess drives R-loop-associated replication stress and HR defects.

    Evidence S9.6 R-loop immunofluorescence, BRCA1 Co-IP with transcription machinery, knockdown/rescue, DNA damage and replication stress assays

    PMID:29513652

    Open questions at the time
    • Direct RNA/DNA species bound by EWSR1 at R-loops not fully defined
    • How EWSR1 dislodges BRCA1 from elongation machinery unresolved
  19. 2015 High

    Showed EWSR1 stabilizes PGC-1α by limiting FBXW7-mediated degradation, linking the protein to mitochondrial biogenesis in multiple tissues.

    Evidence Ews knockout mice, ubiquitination assay, proteasome and Fbxw7 knockdown rescue, mitochondrial measurements

    PMID:25918410

    Open questions at the time
    • Direct molecular step by which EWSR1 limits FBXW7 unknown
    • Whether EWSR1 acts on PGC-1α transcription or protein directly not fully separated
  20. 2019 High

    Identified deubiquitinase- and ligase-based control of fusion protein abundance, establishing EWS-FLI1 stability as a druggable dependency.

    Evidence siRNA/CRISPR screens, Co-IP, ubiquitination and phosphorylation assays, xenografts (USP19, TRIM8, SPOP/OTUD7A)

    PMID:30700749 PMID:34060252 PMID:34329586

    Open questions at the time
    • Whether these enzymes also regulate wild-type EWSR1 partly unresolved
    • Interplay among the competing ligases/DUBs not integrated
  21. 2021 Medium

    Linked the low-complexity domain to formation of cellular protein assemblies required for the fusion's broad partner network including RNA Pol II, bridging biochemistry to gene regulation.

    Evidence cross-linking assembly assay, domain deletion, siRNA knockdown, RNA-seq

    PMID:34035145

    Open questions at the time
    • Whether assemblies are bona fide phase-separated condensates not proven here
    • Composition of assemblies not exhaustively defined
  22. 2023 Medium

    Defined EWSR1 as a centromere identity factor acting through CENP-A binding via SYGQ2 and R-loop binding via its RRM, connecting its disordered and RNA-binding modules to chromosome segregation.

    Evidence Co-IP, CENP-A ChIP after depletion, in vitro R-loop binding, domain mutation analysis, immunofluorescence

    PMID:37243594

    Open questions at the time
    • Mechanism by which EWSR1 retains CENP-A not resolved
    • In vivo significance for chromosome segregation not fully tested
  23. 2024 Medium

    Provided molecular detail that tyrosine residues drive phase separation of the EWS low-complexity domain, defining the sequence determinants of its condensate behavior.

    Evidence PRE-NMR, microscopy, all-atom MD simulations, tyrosine mutational analysis

    PMID:38492239

    Open questions at the time
    • Cellular function of these condensates not directly tested
    • Link to transactivation strength of full-length protein not established

Open questions

Synthesis pass · forward-looking unresolved questions
  • How arginine methylation, tyrosine-driven phase separation, and the competing ubiquitination/deubiquitination network are integrated to coordinate EWSR1's transcription, splicing, centromere, R-loop, and mitochondrial functions remains unresolved.
  • No unified model linking modifications to specific functional outputs
  • Substrate/target specificity of wild-type vs fusion incompletely separated
  • Structural basis of higher-order assembly in cells undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0003723 RNA binding 4 GO:0140110 transcription regulator activity 3 GO:0003677 DNA binding 2 GO:0045182 translation regulator activity 2 GO:0060090 molecular adaptor activity 2
Localization
GO:0005634 nucleus 3 GO:0005856 cytoskeleton 2 GO:0005886 plasma membrane 2 GO:0005694 chromosome 1
Pathway
R-HSA-1640170 Cell Cycle 3 R-HSA-1643685 Disease 2 R-HSA-74160 Gene expression (Transcription) 2 R-HSA-8953854 Metabolism of RNA 2 R-HSA-73894 DNA Repair 1
Partners
AURKBDDX5DHX9FLI1HNRNPKLMNAPRPF6REST

Evidence

Reading pass · 33 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1993 EWS-FLI1 chimeric protein functions as a transcriptional activator; deletion analysis revealed that the EWS N-terminal domain (NTD-EWS) acts as a modulatory/regulatory domain for the transcriptional activation properties of the C-terminal FLI1 activation domain of EWS-FLI1, not as an autonomous activator. Transcriptional reporter assays and deletion analysis in cell transfection experiments Cancer research Medium 7503813
1994 EWS-FLI1 protein displays the same DNA-binding specificity and affinity as wild-type FLI1, with the consensus binding site ACCGGAAG/aT/c; the Ets domain is necessary and sufficient for the DNA-binding specificity of the fusion protein. Epitope-tagged protein binding site selection, truncation mutant DNA-binding assays in vitro The Journal of biological chemistry Medium 7517940
2001 EWS and EWS-FLI1 interact with SF1 and U1C, essential components of the splicing machinery; EWS-FLI1 (but not EWS alone) interferes with hnRNP A1-dependent 5'-splice site selection in an E1A in vivo splicing assay, and this splicing-altering activity coincides with transforming activity. In vivo splicing assay (E1A gene), mutational analysis, protein interaction assays The Journal of biological chemistry Medium 11301318
2001 EWS-FLI1 activates oncogenic pathways independent of its ETS DNA-binding domain (DBD); DBD point mutants and large deletions retain tumor acceleration in NIH 3T3 cells in vivo, while losing DNA binding in vitro, demonstrating DBD-independent oncogenic signaling. In vivo tumor assay in NIH 3T3 / SCID mice, dominant-negative FLI1 constructs, in vitro DNA-binding assays The Journal of biological chemistry Medium 11553628
2001 EWS protein is extensively and asymmetrically dimethylated on arginine residues within its RGG motifs (29 of 30 Arg-Gly sites at least partially methylated); the protein is also present on the cell surface in addition to the nucleus and cytosol. Cell-surface biotinylation, isoelectric focusing, avidin-agarose extraction, MALDI and nanoelectrospray tandem mass spectrometry of in-gel-digested peptides The Journal of biological chemistry High 11278906
2003 EWS protein self-associates through its C-terminal RNA-binding domain in an RNA-dependent manner (sensitive to RNaseA); EWS-FLI1 can also self-associate and interact with FLI1 via its C-terminal FLI1 domain in an RNA-independent manner; the EWS N-terminal domain mediates both homotypic and heterotypic interactions of EWS and EWS-FLI1. FRET, mammalian two-hybrid assay, GST pull-down, immunoprecipitation, RNaseA sensitivity assays Oncogene High 14534527
2003 Methylation process controls EWS protein expression and subcellular localization: inhibition of methylation with adenosine dialdehyde decreases EWS protein in both the nucleus and cell surface; mitogenic stimulation of normal T cells increases methylated EWS on the cell surface ~10-fold. Cell-surface biotinylation, immunoblotting after methylation inhibitor treatment, mitogenic stimulation of PBMC Experimental cell research Medium 12915128
2004 EWS-FLI1 purified as recombinant protein from E. coli adopts a largely unfolded conformation under native conditions, yet specifically binds DNA and activates transcription, confirming its intrinsically disordered nature and transcriptional activity. Recombinant protein purification, circular dichroism, fluorescence spectroscopy, in vitro DNA-binding and transcription assays Biochemistry Medium 15491164
2006 EWS-FLI1 directly binds RNA helicase A (RHA) at the region spanning amino acids 630–1020 of RHA; endogenous RHA co-immunoprecipitates with EWS-FLI1 in ESFT cell lines; RHA and EWS-FLI1 co-occupy target gene promoters (e.g., Id2) by ChIP; RHA stimulates EWS-FLI1 transcriptional activity and enhances anchorage-independent transformation. Phage display, GST pull-down, ELISA, reciprocal Co-IP, chromatin immunoprecipitation, luciferase reporter assay, anchorage-independent growth assay Cancer research High 16740692
2007 EWS is required for completion of B cell development and meiosis in vivo: Ews-null mice show cell-autonomous defects in pre-B lymphocyte development, spermatocyte XY bivalent formation failure, and massive apoptosis during meiosis. Loss of EWS also results in premature senescence of mouse embryonic fibroblasts and hypersensitivity to ionizing radiation. EWS interacts with lamin A/C and its loss reduces lamin A/C expression. Ews knockout mice generation, lymphocyte analysis, meiosis analysis, MEF senescence assays, ionizing radiation sensitivity assay, co-immunoprecipitation (EWS–lamin A/C interaction), immunoblotting The Journal of clinical investigation High 17415412
2007 EWSR1 maintains mitotic integrity: morpholino knockdown of zebrafish ewsr1a/ewsr1b causes multipolar or abnormal mitotic spindles followed by p53-mediated apoptosis in the CNS; siRNA silencing of EWSR1 in HeLa cells causes mitotic defects and apoptosis, confirming conservation of this function. Morpholino knockdown in zebrafish, siRNA knockdown in HeLa cells, mitotic spindle imaging PloS one Medium 17912356
2007 EWS-ATF1 fusion protein, when expressed from the EWS promoter in mice, directly induces FOS expression in an ERK-independent manner; EWS/ATF1 expression is sufficient to transform neural crest-derived cells and produce CCS-like sarcomas in vivo. Inducible transgenic mouse model, lineage-tracing experiments, siRNA knockdown of FOS, promoter analysis The Journal of clinical investigation Medium 23281395
2009 The EWSR1/NR4A3 fusion protein directly activates the PPARG promoter through a specific DNA response element; band-shift experiments confirm EWSR1/NR4A3 binding to this element, and a truncated native NR4A3 isoform can negatively regulate the fusion protein's activity on this promoter. Expression microarray, western blot/IHC validation, band-shift (EMSA) assays, transient transfection reporter assays, bioinformatic promoter analysis The Journal of pathology Medium 18855877
2010 Endogenous EWS is required for hematopoietic stem cell quiescence; Ews-deficient hematopoietic stem/progenitor cells undergo early senescence with increased p16INK4a and senescence-associated β-galactosidase activity. Ews knockout mice, flow cytometry of HSPCs, β-galactosidase senescence assay, p16INK4a immunoblotting Blood Medium 21030557
2012 Acetylation of the C-terminal FLI1 DNA-binding domain of EWS-FLI1 increases its DNA-binding activity in vitro; overexpression of PCAF or treatment with HDAC inhibitors increases EWS-FLI1 transcriptional activity in co-transfection assays. In vitro acetylation assay, EMSA, luciferase reporter assay, PCAF overexpression, HDAC inhibitor treatment Frontiers in oncology Medium 22973553
2013 EWS regulates expression of Drosha (a miRNA microprocessor): EWS deficiency increases Drosha expression, elevating miR-29b and miR-18b levels, which post-transcriptionally repress Col4a1 and CTGF, contributing to dermal developmental defects. Knockdown of Drosha rescues miRNA-dependent downregulation of these targets. Ews knockout mouse embryonic fibroblasts, qPCR for miRNA levels, Drosha knockdown rescue experiment, immunoblotting for Col4a1 and CTGF Cell death and differentiation Medium 24185621
2013 Wild-type EWS interacts directly with REST (RE1-Silencing Transcription Factor) by co-immunoprecipitation; genome-wide ChIP shows EWS binds chromatin at/near NRSE (neuron-restrictive silencer element) sites; EWS and REST cooperatively suppress neuronal gene expression and oncogenic transformation in Ewing sarcoma cells. Co-immunoprecipitation, ChIP-seq, RNA-seq after RNAi, functional transformation assays Genes & cancer Medium 24069508
2014 EWS-FLI1 reprograms gene regulatory circuits by two divergent mechanisms: (1) at GGAA repeat elements EWS-FLI1 multimers induce chromatin opening and create de novo enhancers that physically interact with target promoters; (2) at conserved ETS-motif enhancers EWS-FLI1 displaces wild-type ETS factors to inactivate them. ChIP-seq, ATAC-seq/DNaseI-seq, chromatin conformation capture, EWS-FLI1 knockdown/rescue, luciferase reporter assays Cancer cell High 25453903
2014 EWS-WT1(+KTS) fusion protein directly binds the sequence 5'-GGAGG(A/G)-3' upstream of the LRRC15 gene and transactivates it; this binding site differs from known WT1 consensus sites, demonstrating that the +KTS insertion abrogates canonical WT1 binding but confers a new specificity. cDNA subtractive hybridization, in vitro and in vivo DNA binding assays, ChIP, mutagenesis of binding element, inducible expression system Genes & development High 12923058
2014 EWS-WT1 directly binds the proximal promoter of ASCL1 through multiple WT1-responsive elements and activates ASCL1 transcription, inducing a neural gene expression program and partial neural differentiation in DSRCT cells. Transgenic mouse model (EWS-WT1 under native Ews promoter), promoter-binding assays, ASCL1 knockdown, inducible expression in primary fibroblasts Cancer research Medium 24934812
2014 EWS interacts with Aurora B kinase via the R565 residue in its RGG3 domain, and EWS is required for recruiting Aurora B to the midzone during anaphase; loss of EWS or EWS-FLI1 expression causes midzone defects and aneuploidy; ectopic EWS expression rescues midzone defects in Ewing sarcoma cells. siRNA knockdown, immunofluorescence/live imaging of midzone, co-immunoprecipitation, domain deletion/point mutation analysis, rescue experiments Cell cycle Medium 25483190
2015 EWS-FLI1 acts as a splicing regulatory hub: it binds RNA at intron-exon boundaries (CLIP-seq), interacts with spliceosomal proteins DDX5, hnRNP K, and PRPF6, and produces alternative splicing of CLK1, CASP3, PPFIBP1, and TERT isoforms; the small molecule YK-4-279 disrupts EWS-FLI1 interactions with DDX5 and RHA, reversing splicing alterations. CLIP-seq, exon array, RNA-seq, Co-IP with splicing factors, YK-4-279 inhibitor treatment, telomerase activity assay Proceedings of the National Academy of Sciences of the United States of America High 25737553
2015 EWS-FLI1 inhibits RNA helicase A (RHA) helicase activity in a dose-dependent manner without affecting ATPase activity; EWS-FLI1 has RNA-binding activity and alters the RNA-binding profile of RHA; the (S)-enantiomer of YK-4-279 specifically reverses EWS-FLI1 inhibition of RHA helicase activity. In vitro helicase activity assay, ATPase assay, RNA-binding assay, separated enantiomer treatment Nucleic acids research Medium 25564528
2015 Loss of EWS leads to rapid proteasomal degradation of PGC-1α via increased FBXW7 E3 ubiquitin ligase expression; EWS inactivation causes significant reduction in mitochondrial abundance and activity in MEFs, brown fat, and skeletal muscle; complementation of EWS restores PGC-1α and mitochondrial abundance; depletion of Fbxw7 in Ews-null cells restores PGC-1α. Ews knockout mice, ubiquitination assay, proteasome inhibitor rescue, Fbxw7 knockdown rescue, mitochondrial abundance/activity measurement, immunoblotting Proceedings of the National Academy of Sciences of the United States of America High 25918410
2018 EWSR1 suppresses R-loops and promotes homologous recombination in the transcriptional response to DNA damage; in Ewing sarcoma, EWS-FLI1 increases transcription causing R-loop accumulation and replication stress, and impairs BRCA1-mediated homologous recombination by enriching BRCA1 interactions with the elongating transcription machinery. R-loop immunofluorescence (S9.6 antibody), BRCA1 Co-IP with transcription machinery components, EWSR1 knockdown/rescue, DNA damage assays, replication stress assays Nature High 29513652
2019 EWS-FLI1 modulates ARID1A pre-mRNA splicing to preferentially produce the ARID1A-L isoform; ARID1A-L directly interacts with EWS-FLI1 protein; ARID1A-L promotes Ewing sarcoma growth and reciprocally stabilizes EWS-FLI1 protein, forming a feed-forward oncogenic loop. Co-immunoprecipitation (EWS-FLI1 / ARID1A-L), RNA-seq for isoform analysis, shRNA knockdown, rescue experiments, protein stability assays Nucleic acids research Medium 31392992
2021 TRIM8 is an E3 ubiquitin ligase that ubiquitinates and degrades EWS-FLI1; TRIM8 knockout leads to increased EWS-FLI1 protein levels that is not tolerated by Ewing sarcoma cells; EWS-FLI1 acts as a neomorphic substrate for TRIM8, defining a selective dependency. CRISPR-Cas9 screen, ubiquitination assay, TRIM8 KO/rescue, protein level analysis, selective dependency validation across >700 cancer cell lines Cancer cell High 34329586
2021 SPOP E3 ubiquitin ligase and OTUD7A deubiquitinase control EWS-FLI1 protein stability: casein kinase 1-mediated phosphorylation of the VTSSS degron in the FLI1 domain enhances SPOP-mediated degradation; OTUD7A deubiquitinates and stabilizes EWS-FLI1; depletion of OTUD7A reduces EWS-FLI1 levels and impedes tumor growth in vitro and in vivo. siRNA/shRNA depletion, ubiquitination assays, Co-IP, phosphorylation assays, xenograft mouse models, AI-based drug screening Advanced science Medium 34060252
2019 USP19 deubiquitinase binds to the N-terminal EWS region of EWS-FLI1 and stabilizes the fusion protein; depletion of USP19 reduces EWS-FLI1 protein levels, decreases cell growth, and delays tumor growth in vivo; stabilization is specific for the fusion protein (not EWSR1 or FLI1 alone). siRNA screening, Co-IP, ubiquitination assays, stable shRNA depletion, xenograft tumor assay Scientific reports Medium 30700749
2021 EWS-FLI1 is incorporated into a protein granule/assembly in cells via its low-complexity (LC) domain; the LC domain is required for EWS-FLI1 to form these assemblies and interact with its broad network of protein partners including RNA Pol II; EWSR1 knockdown affects a larger than expected set of transcripts, including many EWS-FLI1-regulated genes. Cross-linking-based protein assembly assay, siRNA-mediated knockdown, RNA-seq, domain deletion analysis RNA Medium 34035145
2022 EWSR1-ATF1 displays distinct DNA binding that requires the EWSR1 domain and promotes ATF1 retargeting to new distal chromatin sites, activating a 3D connectivity network controlling oncogenic and differentiation programs in Clear Cell Sarcoma; EWSR1-ATF1 depletion reconfigures 3D connectivity and promotes neural crest developmental programs. ChIP-seq, Hi-C/3D chromatin conformation capture, ATAC-seq, EWSR1-ATF1 depletion, CUT&RUN Nature communications Medium 35477713
2023 EWSR1 maintains centromere identity by interacting with CENP-A through its SYGQ2 prion-like domain region; EWSR1 binds R-loops through its RNA-recognition motif in vitro; both the SYGQ2 domain and RNA-recognition motif are required for EWSR1 to maintain CENP-A at the centromere in interphase cells. Co-immunoprecipitation (CENP-A–EWSR1), CENP-A ChIP after EWSR1 depletion, in vitro R-loop binding assay, domain deletion/mutation analysis, immunofluorescence Cell reports Medium 37243594
2024 The intrinsically disordered low-complexity domain of EWS (EWSLCD) undergoes phase separation/condensate formation driven by tyrosine residues; higher density and proximity of tyrosines amplify condensate formation; MD simulations revealed tyrosine-rich termini adopt compact conformations with unique intramolecular and intermolecular contact networks. Paramagnetic relaxation enhancement NMR, microscopy (phase separation), all-atom molecular dynamics simulations, mutational analysis of tyrosine residues Journal of the American Chemical Society Medium 38492239

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 EWS-FLI1 utilizes divergent chromatin remodeling mechanisms to directly activate or repress enhancer elements in Ewing sarcoma. Cancer cell 363 25453903
2001 Biology of EWS/ETS fusions in Ewing's family tumors. Oncogene 277 11607824
2022 EWSR1-induced circNEIL3 promotes glioma progression and exosome-mediated macrophage immunosuppressive polarization via stabilizing IGF2BP3. Molecular cancer 263 35031058
2018 EWS-FLI1 increases transcription to cause R-loops and block BRCA1 repair in Ewing sarcoma. Nature 261 29513652
1996 Fusion of the EWS and CHOP genes in myxoid liposarcoma. Oncogene 237 8637704
2012 Tumors with EWSR1-CREB1 and EWSR1-ATF1 fusions: the current status. The American journal of surgical pathology 195 22510762
2007 EWSR1-CREB1 and EWSR1-ATF1 fusion genes in angiomatoid fibrous histiocytoma. Clinical cancer research : an official journal of the American Association for Cancer Research 168 18094413
1993 EWS/Fli-1 chimeric protein is a transcriptional activator. Cancer research 159 7503813
1994 EWS-erg and EWS-Fli1 fusion transcripts in Ewing's sarcoma and primitive neuroectodermal tumors with variant translocations. The Journal of clinical investigation 139 8040301
2005 EWS-ETS oncoproteins: the linchpins of Ewing tumors. Gene 136 16202544
2018 EWS/FLI Confers Tumor Cell Synthetic Lethality to CDK12 Inhibition in Ewing Sarcoma. Cancer cell 134 29358035
2019 Therapeutic targeting of circ-CUX1/EWSR1/MAZ axis inhibits glycolysis and neuroblastoma progression. EMBO molecular medicine 133 31709724
2013 The diversity of soft tissue tumours with EWSR1 gene rearrangements: a review. Histopathology 125 24320889
2010 Oncogenic partnerships: EWS-FLI1 protein interactions initiate key pathways of Ewing's sarcoma. Clinical cancer research : an official journal of the American Association for Cancer Research 123 20547696
2015 Oncogenic fusion protein EWS-FLI1 is a network hub that regulates alternative splicing. Proceedings of the National Academy of Sciences of the United States of America 121 25737553
2010 Soft tissue tumors associated with EWSR1 translocation. Virchows Archiv : an international journal of pathology 119 19936782
2007 Ewing sarcoma gene EWS is essential for meiosis and B lymphocyte development. The Journal of clinical investigation 119 17415412
2014 Recurrent EWSR1-CREB3L1 gene fusions in sclerosing epithelioid fibrosarcoma. The American journal of surgical pathology 117 24441665
2001 Loss of p16 pathways stabilizes EWS/FLI1 expression and complements EWS/FLI1 mediated transformation. Oncogene 115 11709708
2006 Oncoprotein EWS-FLI1 activity is enhanced by RNA helicase A. Cancer research 113 16740692
1994 The FLI-1 and chimeric EWS-FLI-1 oncoproteins display similar DNA binding specificities. The Journal of biological chemistry 111 7517940
2020 EWSR1/FUS-CREB fusions define a distinctive malignant epithelioid neoplasm with predilection for mesothelial-lined cavities. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 100 32770123
2005 The EWS-WT1 gene fusion in desmoplastic small round cell tumor. Seminars in cancer biology 99 15826834
2021 TRIM8 modulates the EWS/FLI oncoprotein to promote survival in Ewing sarcoma. Cancer cell 94 34329586
2019 Clinical, pathological, and genomic features of EWSR1-PATZ1 fusion sarcoma. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 92 31189996
2001 EWS/FLI alters 5'-splice site selection. The Journal of biological chemistry 84 11301318
2018 EWSR1, a multifunctional protein, regulates cellular function and aging via genetic and epigenetic pathways. Biochimica et biophysica acta. Molecular basis of disease 78 30481590
2001 Exposure on cell surface and extensive arginine methylation of ewing sarcoma (EWS) protein. The Journal of biological chemistry 77 11278906
2005 Ewing's sarcoma oncoprotein EWS-FLI1: the perfect target without a therapeutic agent. Future oncology (London, England) 72 16556028
2015 Novel FUS-KLF17 and EWSR1-KLF17 fusions in myoepithelial tumors. Genes, chromosomes & cancer 71 25706482
2017 EWSR1 fusion proteins mediate PAX7 expression in Ewing sarcoma. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 70 28643791
2019 Spindle and Round Cell Sarcoma With EWSR1-PATZ1 Gene Fusion: A Sarcoma With Polyphenotypic Differentiation. The American journal of surgical pathology 68 30379650
2013 Molecular detection and targeting of EWSR1 fusion transcripts in soft tissue tumors. Medical oncology (Northwood, London, England) 66 23329308
2013 RNA sequencing identifies fusion of the EWSR1 and YY1 genes in mesothelioma with t(14;22)(q32;q12). Genes, chromosomes & cancer 63 23630070
2018 EWSR1-PATZ1 gene fusion may define a new glioneuronal tumor entity. Brain pathology (Zurich, Switzerland) 62 29679497
2021 SPOP and OTUD7A Control EWS-FLI1 Protein Stability to Govern Ewing Sarcoma Growth. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 60 34060252
2019 DNA methylation profiling distinguishes Ewing-like sarcoma with EWSR1-NFATc2 fusion from Ewing sarcoma. Journal of cancer research and clinical oncology 60 30895378
2016 Therapeutic opportunities in Ewing sarcoma: EWS-FLI inhibition via LSD1 targeting. Oncotarget 60 26848860
2019 EWSR1-NFATC2 and FUS-NFATC2 Gene Fusion-Associated Mesenchymal Tumors: Clinicopathologic Correlation and Literature Review. Sarcoma 57 31049020
2018 EWS/ETS-Driven Ewing Sarcoma Requires BET Bromodomain Proteins. Cancer research 57 29898995
2019 Mesenchymal Tumors with EWSR1 Gene Rearrangements. Surgical pathology clinics 54 30709442
2009 The EWSR1/NR4A3 fusion protein of extraskeletal myxoid chondrosarcoma activates the PPARG nuclear receptor gene. The Journal of pathology 53 18855877
2001 DNA binding domain-independent pathways are involved in EWS/FLI1-mediated oncogenesis. The Journal of biological chemistry 53 11553628
2007 Ewing sarcoma protein ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo. PloS one 52 17912356
2019 EWS-FLI1 modulated alternative splicing of ARID1A reveals novel oncogenic function through the BAF complex. Nucleic acids research 51 31392992
2004 Recombinant EWS-FLI1 oncoprotein activates transcription. Biochemistry 51 15491164
2024 Insights into Molecular Diversity within the FUS/EWS/TAF15 Protein Family: Unraveling Phase Separation of the N-Terminal Low-Complexity Domain from RNA-Binding Protein EWS. Journal of the American Chemical Society 49 38492239
2015 EWS/FLI1 Target Genes and Therapeutic Opportunities in Ewing Sarcoma. Frontiers in oncology 49 26258070
2016 Targeting the EWS-ETS transcriptional program by BET bromodomain inhibition in Ewing sarcoma. Oncotarget 48 26623725
2019 Trabectedin Inhibits EWS-FLI1 and Evicts SWI/SNF from Chromatin in a Schedule-dependent Manner. Clinical cancer research : an official journal of the American Association for Cancer Research 46 30723142
1995 EWS-FLI-1 and EWS-ERG chimeric mRNAs in Ewing's sarcoma and primitive neuroectodermal tumor. International journal of cancer 46 7591257
2003 Identification of a DNA-binding site and transcriptional target for the EWS-WT1(+KTS) oncoprotein. Genes & development 45 12923058
2010 Ewing sarcoma gene Ews regulates hematopoietic stem cell senescence. Blood 44 21030557
2013 EWS/ATF1 expression induces sarcomas from neural crest-derived cells in mice. The Journal of clinical investigation 43 23281395
2008 EWS-Fli1 up-regulates expression of the Aurora A and Aurora B kinases. Molecular cancer research : MCR 43 19074838
2003 Homotypic and heterotypic interactions of EWS, FLI1 and their oncogenic fusion protein. Oncogene 43 14534527
2022 EWS/FLI mediated reprogramming of 3D chromatin promotes an altered transcriptional state in Ewing sarcoma. Nucleic acids research 42 36124657
2023 ETV6 dependency in Ewing sarcoma by antagonism of EWS-FLI1-mediated enhancer activation. Nature cell biology 41 36658219
2013 What the EWSR1-ATF1 fusion has taught us about hyalinizing clear cell carcinoma. Head and neck pathology 41 23459838
2019 Cutaneous Syncytial Myoepithelioma Is Characterized by Recurrent EWSR1-PBX3 Fusions. The American journal of surgical pathology 40 31135487
2022 EWSR1-ATF1 dependent 3D connectivity regulates oncogenic and differentiation programs in Clear Cell Sarcoma. Nature communications 37 35477713
2015 RNA helicase A activity is inhibited by oncogenic transcription factor EWS-FLI1. Nucleic acids research 37 25564528
2013 A multifunctional protein EWS regulates the expression of Drosha and microRNAs. Cell death and differentiation 36 24185621
2002 EWS-FLI1 and Ewing's sarcoma: recent molecular data and new insights. Cancer biology & therapy 36 12432241
2021 Identification of EWSR1-NFATC2 fusion in simple bone cysts. Histopathology 35 33316098
2014 EWS-WT1 oncoprotein activates neuronal reprogramming factor ASCL1 and promotes neural differentiation. Cancer research 35 24934812
2020 EWS-FLI1 regulates and cooperates with core regulatory circuitry in Ewing sarcoma. Nucleic acids research 34 33080033
2019 USP19 deubiquitinates EWS-FLI1 to regulate Ewing sarcoma growth. Scientific reports 32 30700749
2004 Role of protein-protein interactions in the antiapoptotic function of EWS-Fli-1. Oncogene 32 15273724
2015 Inactivation of EWS reduces PGC-1α protein stability and mitochondrial homeostasis. Proceedings of the National Academy of Sciences of the United States of America 31 25918410
2021 One oncogene, several vulnerabilities: EWS/FLI targeted therapies for Ewing sarcoma. Journal of bone oncology 30 34976713
2016 Englerin A Inhibits EWS-FLI1 DNA Binding in Ewing Sarcoma Cells. The Journal of biological chemistry 30 26961871
2003 Expression and subcellular localization of Ewing sarcoma (EWS) protein is affected by the methylation process. Experimental cell research 30 12915128
2020 RING1B recruits EWSR1-FLI1 and cooperates in the remodeling of chromatin necessary for Ewing sarcoma tumorigenesis. Science advances 29 33097530
2022 EWS::FLI1 and HOXD13 Control Tumor Cell Plasticity in Ewing Sarcoma. Clinical cancer research : an official journal of the American Association for Cancer Research 28 35653119
2021 Fusion protein EWS-FLI1 is incorporated into a protein granule in cells. RNA (New York, N.Y.) 27 34035145
2013 Cutaneous neoplasms showing EWSR1 rearrangement. Advances in anatomic pathology 27 23399793
2008 EWS/FLI1 suppresses retinoblastoma protein function and senescence in Ewing's sarcoma cells. Journal of orthopaedic research : official publication of the Orthopaedic Research Society 27 18271016
2015 EWS and FUS bind a subset of transcribed genes encoding proteins enriched in RNA regulatory functions. BMC genomics 26 26573619
2011 mRNA and protein levels of FUS, EWSR1, and TAF15 are upregulated in liposarcoma. Genes, chromosomes & cancer 26 21344536
2022 Therapeutic targeting the oncogenic driver EWSR1::FLI1 in Ewing sarcoma through inhibition of the FACT complex. Oncogene 23 36357572
2023 Regulation of EWSR1-FLI1 Function by Post-Transcriptional and Post-Translational Modifications. Cancers 22 36672331
2022 EWSR1-TFCP2 in an adolescent represents an extremely rare and aggressive form of intraosseous spindle cell rhabdomyosarcomas. Cold Spring Harbor molecular case studies 22 35768243
2019 EWSR1-FLI1 Activation of the Cancer/Testis Antigen FATE1 Promotes Ewing Sarcoma Survival. Molecular and cellular biology 22 31036566
2016 EWSR1/ELF5 induces acute myeloid leukemia by inhibiting p53/p21 pathway. Cancer science 22 27627705
2023 EWS/FLI1 Characterization, Activation, Repression, Target Genes and Therapeutic Opportunities in Ewing Sarcoma. International journal of molecular sciences 21 37894854
2023 EWSR1 maintains centromere identity. Cell reports 20 37243594
2015 PI3K/AKT signaling modulates transcriptional expression of EWS/FLI1 through specificity protein 1. Oncotarget 20 26336820
2012 Acetylation Increases EWS-FLI1 DNA Binding and Transcriptional Activity. Frontiers in oncology 20 22973553
2022 MANF/EWSR1/ANXA6 pathway might as the bridge between hypolipidemia and major depressive disorder. Translational psychiatry 19 36585419
2013 EWS and RE1-Silencing Transcription Factor Inhibit Neuronal Phenotype Development and Oncogenic Transformation in Ewing Sarcoma. Genes & cancer 19 24069508
2023 Sarcomas Harboring EWSR1::PATZ1 Fusions: A Clinicopathologic Study of 17 Cases. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 18 38043789
2021 EWSR1-WT1 Target Genes and Therapeutic Options Identified in a Novel DSRCT In Vitro Model. Cancers 18 34885181
2020 High Specificity of BCL11B and GLG1 for EWSR1-FLI1 and EWSR1-ERG Positive Ewing Sarcoma. Cancers 18 32164354
2014 Ewing sarcoma EWS protein regulates midzone formation by recruiting Aurora B kinase to the midzone. Cell cycle (Georgetown, Tex.) 18 25483190
2007 The EWS-Oct-4 fusion gene encodes a transforming gene. The Biochemical journal 18 17564582
2023 Targeted Therapy for EWS-FLI1 in Ewing Sarcoma. Cancers 16 37627063
2021 Challenges in modeling EWS-FLI1-driven transgenic mouse model for Ewing sarcoma. American journal of translational research 16 34956445
2019 EWSR1 translocation in primary hyalinising clear cell carcinoma of the thymus. Histopathology 16 31050844
2023 Targeting the Clear Cell Sarcoma Oncogenic Driver Fusion Gene EWSR1::ATF1 by HDAC Inhibition. Cancer research communications 15 37405123

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