{"gene":"EVC","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2007,"finding":"EVC protein localizes to the base of the primary cilium (basal body) in chondrocytes, and loss of Evc in mice impairs transcriptional activation of Ihh target genes (Ptch1, Gli1) downstream of Smo but upstream of Gli3 processing, establishing EVC as an intracellular component of the hedgehog signal transduction pathway required for normal transcriptional activation.","method":"Immunofluorescence with anti-Evc antibodies; Evc knockout mouse model; in vitro studies with Evc-/- cells; western blot analysis of Gli3 processing; qRT-PCR of Ihh target genes","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KO mouse model with defined cellular phenotype, multiple orthogonal methods (IHC localization, in vitro pathway placement, western blot), independently replicated in later studies","pmids":["17660199"],"is_preprint":false},{"year":2011,"finding":"EVC2 (Evc2) interacts directly with EVC (Evc) as demonstrated by yeast two-hybrid and co-immunoprecipitation; the two proteins co-localize at the basal body and cilia membrane, and their localization to basal body and cilia is mutually co-dependent. EVC2 is a transmembrane protein with an extracellular portion and intracellular C-terminus, while EVC lacks an extracellular portion. Additionally, full-length EVC2 (but not EVC) is also found in the nucleus.","method":"Yeast two-hybrid screen using EVC as bait; co-immunoprecipitation; immunofluorescence with anti-Evc2 antibodies; subcellular fractionation western blot; co-transfection localization studies","journal":"BMC biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction confirmed by yeast-2-hybrid and Co-IP, localization confirmed by multiple orthogonal methods, co-dependency established by transfection rescue experiments","pmids":["21356043"],"is_preprint":false},{"year":2012,"finding":"The EVC-EVC2 complex acts downstream of Smo to transduce Hedgehog signal; Hh stimulation induces phosphorylation-dependent binding of EVC/EVC2 to the Smo C-terminal intracellular tail; this binding is abolished in Kif3a-/- cilium-deficient cells. EVC/EVC2 are dispensable for constitutive Gli activity in Sufu-/- cells, indicating they act upstream of Sufu to promote Gli activation.","method":"Co-immunoprecipitation (Smo with Evc/Evc2); constitutively active Smo constructs; Sufu-/- and Kif3a-/- cell lines; Smo phosphorylation assays; Hh pathway reporter assays","journal":"Cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP with functional validation, multiple genetic backgrounds tested (Sufu-/-, Kif3a-/-), replicated binding finding independently confirmed in companion paper (PMID:23026747)","pmids":["22986504"],"is_preprint":false},{"year":2012,"finding":"EVC and EVC2 are mutually required for localizing to primary cilia and for maintaining their normal protein levels. Smo translocation to the cilium is normal in Evc2-/- chondrocytes after Hh activation, but Gli3 recruitment to cilia tips is reduced and Sufu/Gli3 dissociation is impaired. Smo co-precipitates with EVC/EVC2 complex. A dominantly acting Evc2 mutation (Evc2Δ43, Weyers) causes mislocalization of EVC/EVC2Δ43 within the cilium and reproduces Gli3-related molecular defects. Evc silencing in Sufu-/- cells attenuates Hh pathway output, indicating EVC/EVC2 also promote Hh signaling in the absence of Sufu.","method":"Evc and Evc2 knockout mouse models; Co-immunoprecipitation; immunofluorescence for cilia tip localization; western blot for Sufu/Gli3 dissociation; siRNA silencing in Sufu-/- cells; SAG (Smo agonist) stimulation assays","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple knockout mouse models with defined molecular phenotypes, Co-IP, multiple orthogonal methods, independently replicates and extends PMID:22986504","pmids":["23026747"],"is_preprint":false},{"year":2014,"finding":"EFCAB7 and IQCE form a complex that anchors EVC-EVC2 in a signaling microdomain at the base of primary cilia (EvC zone). EFCAB7 binds directly to a C-terminal disordered region of EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of GLI2, phenocopying the Weyers cellular defect.","method":"Co-immunoprecipitation; immunofluorescence; siRNA depletion of EFCAB7; GLI2 activation assays; domain mapping of EFCAB7-EVC2 interaction","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP with domain mapping, functional rescue/depletion experiments with defined molecular phenotype, multiple orthogonal methods in a single rigorous study","pmids":["24582806"],"is_preprint":false},{"year":2011,"finding":"Evc mediates Hedgehog signaling in osteoblasts (localizing to osteoblast primary cilia) in addition to chondrocytes; Evc promotes chondrocyte proliferation, chondrocyte hypertrophy, and osteoblast differentiation in the perichondrium, implicating it in both Pthrp-dependent and Pthrp-independent Ihh functions. Evc mediates Hh target gene expression in inner perichondrial cells but is dispensable in the external perichondrial layers.","method":"Evc-/- mouse model in defined genetic background; Ptch1-LacZ reporter mice; immunofluorescence for osteoblast cilia localization; qRT-PCR and in situ hybridization of osteoblast and Wnt/β-catenin markers","journal":"Bone","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean KO mouse model with multiple defined cellular phenotypes and marker analysis, direct cilia localization experiment","pmids":["21911092"],"is_preprint":false},{"year":2009,"finding":"Ectopic expression of EVC2 Weyers acrodental dysostosis missense variants (in the final exon of EVC2) in NIH 3T3 cells impairs Hedgehog signal transduction, consistent with a dominant-negative mechanism, whereas expression of a truncated EVC2 protein mimicking an Ellis-van Creveld loss-of-function mutation does not impair Hh signaling in this assay.","method":"Ectopic expression of murine EVC2 Weyers variants in NIH 3T3 cells; Hedgehog pathway reporter assays","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, functional cell-based assay but limited to overexpression without rescue controls","pmids":["19810119"],"is_preprint":false},{"year":2013,"finding":"Evc is required for a spatially and temporally regulated Shh signaling response during molar development; absence of Evc causes progressive loss of Shh pathway response in a buccal-to-lingual direction and is associated with displaced Wnt pathway activity, establishing Evc as necessary for normal buccal-lingual axis patterning during tooth morphogenesis.","method":"Evc knockout mouse model; in situ hybridization for Shh target genes; LacZ reporter analysis for Wnt pathway; histological analysis of molar development","journal":"Journal of dental research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse, multiple pathway markers, defined spatial and temporal phenotype, single lab","pmids":["23315474"],"is_preprint":false},{"year":2015,"finding":"WDR35 (IFT121), a retrograde intraflagellar transport protein, is specifically required for entry of EVC, EVC2, and Smoothened into the ciliary compartment. In Wdr35-/- cells all three proteins fail to localize to cilia, but this defect is not seen in Dync2h1-/- (retrograde motor mutant) cilia, establishing that IFT-A but not the retrograde IFT motor is required for EVC/EVC2/SMO ciliary targeting.","method":"Wdr35-/- and Dync2h1-/- mouse fibroblast cell lines; immunofluorescence for EVC, EVC2, and SMO cilia localization; Hedgehog pathway activity assays; expression of Wdr35 disease cDNAs for rescue","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple mutant cell lines compared, rescue by cDNA re-expression, multiple protein targets analyzed by direct imaging, mechanistic distinction between IFT-A and retrograde motor roles established","pmids":["25908617"],"is_preprint":false},{"year":2023,"finding":"The EVC-EVC2 complex is subject to ubiquitination and SUMOylation of its cytosolic tails. Monoubiquitination of EVC-EVC2 cytosolic tails reduces protein levels. SUMOylation with SUMO3 enhances EVC-EVC2 accumulation at the EvC zone at the base of cilia, likely via increased binding to the EFCAB7-IQCE complex. A second EFCAB7-binding motif in EVC2's Weyers-deleted peptide was mapped, in addition to the previously characterized one. Known interactors EVC2, IQCE, EFCAB7 were confirmed, and USP7 (a deubiquitinase involved in Hh signaling) was identified as a new interactor, though ubiquitination of EVC-EVC2 was found to be independent of USP7 and USP48.","method":"Endogenous EVC protein interactome by mass spectrometry in control and Evc-null cells; ubiquitination assays; SUMO modification assays; immunofluorescence for EvC zone localization; domain mapping of EFCAB7-binding motifs","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteomic interactome screen plus direct modification assays and localization imaging, but single lab and some findings still require independent replication","pmids":["37576597"],"is_preprint":false},{"year":2020,"finding":"Hypomorphic EVC missense mutations (p.Arg663Pro and splice variant c.1316-7A>G) produce proteins that retain partial ability to complex with EVC2, whereas the p.Arg622Ter nonsense mutation is a complete loss-of-function allele that cannot complex with EVC2, establishing that the degree of EVC-EVC2 complex formation correlates with clinical severity.","method":"Patient-derived fibroblasts; complementary DNA sequencing; immunoblot; immunofluorescence; Evc-/- mouse embryonic fibroblast complementation assays","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — patient fibroblasts with multiple orthogonal methods and mouse MEF complementation, single lab","pmids":["32906221"],"is_preprint":false},{"year":2025,"finding":"EVC protein facilitates Shh signaling in notochord-derived nucleus pulposus cells; loss of EVC reduces Gli3 processing, impairs Shh pathway activity, and alters extracellular matrix organization toward fibrosis. TGF-β signaling suppresses EVC expression, indicating crosstalk between TGF-β and Hedgehog pathways at the level of EVC.","method":"Evc knockout mouse model; CRISPR-engineered human NP cells; proteome profiling; Gli3 processing western blot; ECM composition analysis; TGF-β treatment assays","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two model systems (mouse KO and CRISPR human cells), multiple pathway readouts, single lab","pmids":["41550739"],"is_preprint":false},{"year":2025,"finding":"EVC and EVC2 are aberrantly overexpressed in a subset of AMLs (particularly those with ASXL1 mutations or t(8;21) translocation); functional loss of EVC/EVC2 impairs leukemia cell proliferation, promotes differentiation, and blocks AML progression in vivo. The leukemogenic role of EVC/EVC2 is mediated through MYC pathway activation, independent of their canonical role in Hedgehog signaling.","method":"shRNA/CRISPR loss-of-function in AML cell lines; in vivo AML transplantation models; ChIP-seq for AML1-ETO occupancy at EVC/EVC2 promoters; chromatin interaction analysis; MYC pathway gene expression analysis","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple functional assays (in vitro and in vivo) with mechanistic pathway placement, single lab, pathway independence from Hh needs further confirmation","pmids":["41249566"],"is_preprint":false},{"year":2004,"finding":"Evc protein is expressed in prehypertrophic and hypertrophic chondrocytes of the epiphyseal growth plate in rat, as determined by immunohistochemistry and in situ hybridization, consistent with a function in chondrocyte differentiation during endochondral ossification.","method":"Immunohistochemistry with anti-Evc antibodies; in situ hybridization; RT-PCR on growth plate tissue","journal":"The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, localization only without direct functional consequence shown","pmids":["15278943"],"is_preprint":false}],"current_model":"EVC encodes a transmembrane ciliary protein that localizes to a signaling microdomain (EvC zone) at the base of primary cilia, where it forms an obligate heterodimeric complex with EVC2; upon Hedgehog pathway activation, phosphorylated Smoothened recruits the EVC-EVC2 complex, which then promotes dissociation of Sufu from Gli3, facilitates Gli3 trafficking to cilia tips, and drives transcriptional activation of Hh target genes—a process regulated by ubiquitination (reducing complex stability) and SUMOylation (enhancing EvC zone accumulation via EFCAB7-IQCE tethering), with ciliary entry of the complex dependent on IFT-A machinery including WDR35."},"narrative":{"mechanistic_narrative":"EVC is a transmembrane ciliary protein that functions as an intracellular transducer of Hedgehog signaling, acting downstream of Smoothened but upstream of Gli3 processing to drive transcriptional activation of Hh target genes such as Ptch1 and Gli1 [PMID:17660199]. It forms an obligate heterodimeric complex with EVC2, the two proteins binding directly and depending on each other for both ciliary localization and protein stability [PMID:21356043, PMID:23026747]. The complex localizes to a signaling microdomain at the base of the primary cilium, the EvC zone, where it is anchored by the EFCAB7-IQCE complex through binding of EFCAB7 to a C-terminal disordered region of EVC2; loss of this tethering mislocalizes EVC-EVC2 and impairs GLI2 activation [PMID:24582806]. Upon Hh activation, phosphorylated Smoothened recruits EVC-EVC2 through its C-terminal tail in a cilium-dependent manner [PMID:22986504], and the complex promotes Sufu/Gli3 dissociation and Gli3 trafficking to cilia tips [PMID:23026747]; EVC-EVC2 also potentiates Hh output independently of Sufu [PMID:22986504, PMID:23026747]. Ciliary entry of EVC, EVC2, and Smoothened requires the IFT-A protein WDR35, distinct from the retrograde IFT motor [PMID:25908617]. The complex is regulated post-translationally, with monoubiquitination of the cytosolic tails reducing protein levels and SUMOylation enhancing EvC zone accumulation via increased EFCAB7-IQCE binding [PMID:37576597]. Through this Hh-transducing role, EVC governs chondrocyte proliferation and hypertrophy and osteoblast differentiation during endochondral ossification [PMID:21911092], tooth buccal-lingual axis patterning [PMID:23315474], and nucleus pulposus extracellular matrix homeostasis [PMID:41550739]. Hypomorphic versus null EVC alleles differ in their ability to complex with EVC2, and the degree of complex formation correlates with the clinical severity of EVC-related skeletal dysplasia [PMID:32906221]; dominant-negative EVC2 variants underlie Weyers acrodental dysostosis [PMID:19810119]. EVC and EVC2 are also aberrantly overexpressed in subsets of AML, where they support leukemic proliferation through MYC pathway activation independent of Hedgehog signaling [PMID:41249566].","teleology":[{"year":2007,"claim":"Established EVC as a bona fide intracellular Hedgehog pathway component by placing its action genetically between Smoothened and Gli3 processing, resolving where in the pathway it operates.","evidence":"Evc knockout mouse and Evc-/- cells with basal body immunofluorescence, Gli3 western blot, and qRT-PCR of Ihh targets","pmids":["17660199"],"confidence":"High","gaps":["Molecular partners mediating transduction not yet identified","Mechanism of basal body targeting unknown"]},{"year":2011,"claim":"Identified EVC2 as the direct, mutually co-dependent partner of EVC, defining the functional unit as a heterodimer rather than a solo protein.","evidence":"Yeast two-hybrid screen with EVC bait, co-immunoprecipitation, immunofluorescence, and subcellular fractionation","pmids":["21356043"],"confidence":"High","gaps":["Stoichiometry and structure of the complex not determined","Functional significance of nuclear full-length EVC2 unexplored"]},{"year":2011,"claim":"Extended EVC's role beyond chondrocytes to osteoblasts and to both Pthrp-dependent and -independent Ihh functions, demonstrating cell-type-specific deployment within skeletal development.","evidence":"Evc-/- mice with Ptch1-LacZ reporter, osteoblast cilia immunofluorescence, and marker analysis","pmids":["21911092"],"confidence":"High","gaps":["Basis for differential requirement across perichondrial layers unresolved"]},{"year":2012,"claim":"Showed that Hh activation drives phosphorylation-dependent binding of EVC-EVC2 to the Smoothened C-terminal tail and that the complex acts upstream of Sufu, mechanistically coupling Smo activation to downstream Gli activation.","evidence":"Co-IP of Smo with Evc/Evc2, constitutively active Smo, and Sufu-/- / Kif3a-/- cell lines with reporter assays","pmids":["22986504"],"confidence":"High","gaps":["Identity of the kinase generating the phospho-dependent binding site not defined"]},{"year":2012,"claim":"Defined the downstream output of EVC-EVC2 as promoting Sufu/Gli3 dissociation and Gli3 ciliary tip recruitment, and showed a dominant Weyers Evc2 mutation mislocalizes the complex and reproduces these defects.","evidence":"Evc and Evc2 knockout mice, Co-IP, cilia tip immunofluorescence, Sufu/Gli3 dissociation western blot, and siRNA in Sufu-/- cells","pmids":["23026747"],"confidence":"High","gaps":["Direct biochemical action on the Sufu-Gli3 complex not reconstituted"]},{"year":2014,"claim":"Revealed how EVC-EVC2 is spatially confined to the EvC zone, identifying the EFCAB7-IQCE complex as the anchor that binds the Weyers-deleted region of EVC2 and is required for GLI2 activation.","evidence":"Co-IP with domain mapping, siRNA depletion of EFCAB7, and GLI2 activation assays","pmids":["24582806"],"confidence":"High","gaps":["How EvC zone confinement mechanistically enables signaling not fully resolved"]},{"year":2015,"claim":"Identified the trafficking route into cilia, showing IFT-A protein WDR35 — but not the retrograde motor — is required for ciliary entry of EVC, EVC2, and SMO.","evidence":"Wdr35-/- vs Dync2h1-/- fibroblasts, cilia localization imaging, pathway assays, and disease cDNA rescue","pmids":["25908617"],"confidence":"High","gaps":["Direct physical interaction between IFT-A and EVC cargo not demonstrated"]},{"year":2023,"claim":"Established post-translational control of the complex, showing monoubiquitination lowers EVC-EVC2 levels while SUMOylation boosts EvC zone accumulation, and mapped a second EFCAB7-binding motif plus a USP7 interaction.","evidence":"Endogenous EVC interactome mass spectrometry in control vs Evc-null cells, ubiquitination/SUMO assays, and localization imaging","pmids":["37576597"],"confidence":"Medium","gaps":["Ubiquitin ligase responsible not identified","Functional role of the USP7 interaction unresolved","Single lab, some findings await replication"]},{"year":2020,"claim":"Connected genotype to phenotype by showing that EVC allele severity tracks with residual ability to complex with EVC2, providing a molecular basis for clinical variability.","evidence":"Patient fibroblasts, immunoblot/immunofluorescence, and Evc-/- MEF complementation assays","pmids":["32906221"],"confidence":"Medium","gaps":["Quantitative threshold of complex formation needed for function not established","Single lab"]},{"year":2009,"claim":"Distinguished the disease mechanisms of two EVC2-linked dysplasias, showing Weyers missense variants act dominant-negatively on Hh signaling while a truncating Ellis-van Creveld-type allele does not in the assay.","evidence":"Ectopic expression of murine EVC2 variants in NIH 3T3 cells with Hh reporter assays","pmids":["19810119"],"confidence":"Medium","gaps":["Overexpression assay without rescue controls","Endogenous-level confirmation lacking"]},{"year":2013,"claim":"Showed EVC is required for spatiotemporal Shh response and buccal-lingual axis patterning in tooth development, broadening its developmental remit.","evidence":"Evc knockout mice with in situ hybridization, Wnt LacZ reporter, and histology of molar development","pmids":["23315474"],"confidence":"Medium","gaps":["Mechanism linking EVC loss to displaced Wnt activity not defined","Single lab"]},{"year":2025,"claim":"Extended EVC function to nucleus pulposus ECM homeostasis and uncovered TGF-beta suppression of EVC, identifying pathway crosstalk affecting Shh output and fibrosis.","evidence":"Evc knockout mice and CRISPR human NP cells with Gli3 western blot, proteome profiling, and TGF-beta treatment","pmids":["41550739"],"confidence":"Medium","gaps":["Mechanism of TGF-beta-mediated EVC repression unknown","Single lab"]},{"year":2025,"claim":"Revealed a non-canonical oncogenic role in AML where EVC/EVC2 overexpression supports leukemic proliferation via MYC, independent of Hedgehog signaling.","evidence":"shRNA/CRISPR loss-of-function in AML lines, in vivo transplantation, AML1-ETO ChIP-seq, and MYC pathway expression analysis","pmids":["41249566"],"confidence":"Medium","gaps":["Direct molecular link between EVC and MYC unresolved","Hh-independence requires further confirmation","Single lab"]},{"year":null,"claim":"How the EVC-EVC2 complex biochemically catalyzes Sufu/Gli3 dissociation, the structure of the assembled complex, and the identity of the enzymes controlling its ubiquitination remain unresolved.","evidence":"Not addressed by current timeline","pmids":[],"confidence":"Low","gaps":["No reconstituted biochemical mechanism for Sufu/Gli3 dissociation","No structural model of the EVC-EVC2 complex","Ubiquitin ligase identity unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,2,3]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[2,3,4]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1,3,4]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2,3]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[5,7,11]}],"complexes":["EVC-EVC2 complex","EvC zone (EVC-EVC2-EFCAB7-IQCE)"],"partners":["EVC2","SMO","EFCAB7","IQCE","USP7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P57679","full_name":"EvC complex member EVC","aliases":["DWF-1","Ellis-van Creveld syndrome protein"],"length_aa":992,"mass_kda":112.0,"function":"Component of the EvC complex that positively regulates ciliary Hedgehog (Hh) signaling. Involved in endochondral growth and skeletal development","subcellular_location":"Cell membrane; Cytoplasm, cytoskeleton, cilium basal body; Cell projection, cilium; Cell projection, cilium membrane","url":"https://www.uniprot.org/uniprotkb/P57679/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EVC","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EVC","total_profiled":1310},"omim":[{"mim_id":"619143","title":"CARDIOACROFACIAL DYSPLASIA 2; CAFD2","url":"https://www.omim.org/entry/619143"},{"mim_id":"619142","title":"CARDIOACROFACIAL DYSPLASIA 1; CAFD1","url":"https://www.omim.org/entry/619142"},{"mim_id":"618123","title":"POLYDACTYLY, POSTAXIAL, TYPE A8; PAPA8","url":"https://www.omim.org/entry/618123"},{"mim_id":"617927","title":"OROFACIODIGITAL SYNDROME XVIII; OFD18","url":"https://www.omim.org/entry/617927"},{"mim_id":"617925","title":"SHORT-RIB THORACIC DYSPLASIA 20 WITH POLYDACTYLY; SRTD20","url":"https://www.omim.org/entry/617925"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Basal body","reliability":"Approved"},{"location":"Primary cilium","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EVC"},"hgnc":{"alias_symbol":["DWF-1","EVC1"],"prev_symbol":[]},"alphafold":{"accession":"P57679","domains":[{"cath_id":"-","chopping":"189-685_692-755","consensus_level":"medium","plddt":90.3965,"start":189,"end":755},{"cath_id":"-","chopping":"775-828_864-909","consensus_level":"high","plddt":86.2043,"start":775,"end":909}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P57679","model_url":"https://alphafold.ebi.ac.uk/files/AF-P57679-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P57679-F1-predicted_aligned_error_v6.png","plddt_mean":73.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EVC","jax_strain_url":"https://www.jax.org/strain/search?query=EVC"},"sequence":{"accession":"P57679","fasta_url":"https://rest.uniprot.org/uniprotkb/P57679.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P57679/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P57679"}},"corpus_meta":[{"pmid":"17660199","id":"PMC_17660199","title":"Evc is a positive mediator of Ihh-regulated bone growth that localises at the base of chondrocyte cilia.","date":"2007","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/17660199","citation_count":156,"is_preprint":false},{"pmid":"23026747","id":"PMC_23026747","title":"The ciliary Evc/Evc2 complex interacts with Smo and controls Hedgehog pathway activity in chondrocytes by regulating Sufu/Gli3 dissociation and Gli3 trafficking in primary cilia.","date":"2012","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23026747","citation_count":94,"is_preprint":false},{"pmid":"22986504","id":"PMC_22986504","title":"Smoothened transduces Hedgehog signal by forming a complex with Evc/Evc2.","date":"2012","source":"Cell research","url":"https://pubmed.ncbi.nlm.nih.gov/22986504","citation_count":86,"is_preprint":false},{"pmid":"17024374","id":"PMC_17024374","title":"Sequencing EVC and EVC2 identifies mutations in two-thirds of Ellis-van Creveld syndrome patients.","date":"2006","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17024374","citation_count":80,"is_preprint":false},{"pmid":"24582806","id":"PMC_24582806","title":"EFCAB7 and IQCE regulate hedgehog signaling by tethering the EVC-EVC2 complex to the base of primary cilia.","date":"2014","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/24582806","citation_count":77,"is_preprint":false},{"pmid":"21356043","id":"PMC_21356043","title":"Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus.","date":"2011","source":"BMC biology","url":"https://pubmed.ncbi.nlm.nih.gov/21356043","citation_count":77,"is_preprint":false},{"pmid":"23220543","id":"PMC_23220543","title":"Novel and recurrent EVC and EVC2 mutations in Ellis-van Creveld syndrome and Weyers acrofacial dyostosis.","date":"2012","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23220543","citation_count":59,"is_preprint":false},{"pmid":"19810119","id":"PMC_19810119","title":"Widening the mutation spectrum of EVC and EVC2: ectopic expression of Weyer variants in NIH 3T3 fibroblasts disrupts Hedgehog signaling.","date":"2009","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/19810119","citation_count":52,"is_preprint":false},{"pmid":"23315474","id":"PMC_23315474","title":"Evc regulates a symmetrical response to Shh signaling in molar development.","date":"2013","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/23315474","citation_count":50,"is_preprint":false},{"pmid":"25908617","id":"PMC_25908617","title":"Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium.","date":"2015","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25908617","citation_count":49,"is_preprint":false},{"pmid":"21911092","id":"PMC_21911092","title":"Evc works in chondrocytes and osteoblasts to regulate multiple aspects of growth plate development in the appendicular skeleton and cranial base.","date":"2011","source":"Bone","url":"https://pubmed.ncbi.nlm.nih.gov/21911092","citation_count":36,"is_preprint":false},{"pmid":"18947413","id":"PMC_18947413","title":"Extending the spectrum of Ellis van Creveld syndrome: a large family with a mild mutation in the EVC gene.","date":"2008","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/18947413","citation_count":22,"is_preprint":false},{"pmid":"29321360","id":"PMC_29321360","title":"Ellis-van Creveld syndrome and profound deafness resulted by sequence variants in the EVC/EVC2 and TMC1 genes.","date":"2017","source":"Journal of 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biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/41714013","citation_count":0,"is_preprint":false},{"pmid":"40475170","id":"PMC_40475170","title":"Ellis-Van Creveld Syndrome with Severe Mitral Valve Insufficiency Caused by a Homozygous Intragenic Deletion of the EVC Gene.","date":"2024","source":"Molecular syndromology","url":"https://pubmed.ncbi.nlm.nih.gov/40475170","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.11.27.690959","title":"Top-down feedback can explain the existence of working memory traces in early visual cortex","date":"2025-11-29","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.27.690959","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.11.03.686222","title":"Predictive vision-language integration in the human visual 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for normal transcriptional activation.\",\n      \"method\": \"Immunofluorescence with anti-Evc antibodies; Evc knockout mouse model; in vitro studies with Evc-/- cells; western blot analysis of Gli3 processing; qRT-PCR of Ihh target genes\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KO mouse model with defined cellular phenotype, multiple orthogonal methods (IHC localization, in vitro pathway placement, western blot), independently replicated in later studies\",\n      \"pmids\": [\"17660199\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EVC2 (Evc2) interacts directly with EVC (Evc) as demonstrated by yeast two-hybrid and co-immunoprecipitation; the two proteins co-localize at the basal body and cilia membrane, and their localization to basal body and cilia is mutually co-dependent. EVC2 is a transmembrane protein with an extracellular portion and intracellular C-terminus, while EVC lacks an extracellular portion. Additionally, full-length EVC2 (but not EVC) is also found in the nucleus.\",\n      \"method\": \"Yeast two-hybrid screen using EVC as bait; co-immunoprecipitation; immunofluorescence with anti-Evc2 antibodies; subcellular fractionation western blot; co-transfection localization studies\",\n      \"journal\": \"BMC biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction confirmed by yeast-2-hybrid and Co-IP, localization confirmed by multiple orthogonal methods, co-dependency established by transfection rescue experiments\",\n      \"pmids\": [\"21356043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The EVC-EVC2 complex acts downstream of Smo to transduce Hedgehog signal; Hh stimulation induces phosphorylation-dependent binding of EVC/EVC2 to the Smo C-terminal intracellular tail; this binding is abolished in Kif3a-/- cilium-deficient cells. EVC/EVC2 are dispensable for constitutive Gli activity in Sufu-/- cells, indicating they act upstream of Sufu to promote Gli activation.\",\n      \"method\": \"Co-immunoprecipitation (Smo with Evc/Evc2); constitutively active Smo constructs; Sufu-/- and Kif3a-/- cell lines; Smo phosphorylation assays; Hh pathway reporter assays\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP with functional validation, multiple genetic backgrounds tested (Sufu-/-, Kif3a-/-), replicated binding finding independently confirmed in companion paper (PMID:23026747)\",\n      \"pmids\": [\"22986504\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EVC and EVC2 are mutually required for localizing to primary cilia and for maintaining their normal protein levels. Smo translocation to the cilium is normal in Evc2-/- chondrocytes after Hh activation, but Gli3 recruitment to cilia tips is reduced and Sufu/Gli3 dissociation is impaired. Smo co-precipitates with EVC/EVC2 complex. A dominantly acting Evc2 mutation (Evc2Δ43, Weyers) causes mislocalization of EVC/EVC2Δ43 within the cilium and reproduces Gli3-related molecular defects. Evc silencing in Sufu-/- cells attenuates Hh pathway output, indicating EVC/EVC2 also promote Hh signaling in the absence of Sufu.\",\n      \"method\": \"Evc and Evc2 knockout mouse models; Co-immunoprecipitation; immunofluorescence for cilia tip localization; western blot for Sufu/Gli3 dissociation; siRNA silencing in Sufu-/- cells; SAG (Smo agonist) stimulation assays\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple knockout mouse models with defined molecular phenotypes, Co-IP, multiple orthogonal methods, independently replicates and extends PMID:22986504\",\n      \"pmids\": [\"23026747\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EFCAB7 and IQCE form a complex that anchors EVC-EVC2 in a signaling microdomain at the base of primary cilia (EvC zone). EFCAB7 binds directly to a C-terminal disordered region of EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of GLI2, phenocopying the Weyers cellular defect.\",\n      \"method\": \"Co-immunoprecipitation; immunofluorescence; siRNA depletion of EFCAB7; GLI2 activation assays; domain mapping of EFCAB7-EVC2 interaction\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP with domain mapping, functional rescue/depletion experiments with defined molecular phenotype, multiple orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"24582806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Evc mediates Hedgehog signaling in osteoblasts (localizing to osteoblast primary cilia) in addition to chondrocytes; Evc promotes chondrocyte proliferation, chondrocyte hypertrophy, and osteoblast differentiation in the perichondrium, implicating it in both Pthrp-dependent and Pthrp-independent Ihh functions. Evc mediates Hh target gene expression in inner perichondrial cells but is dispensable in the external perichondrial layers.\",\n      \"method\": \"Evc-/- mouse model in defined genetic background; Ptch1-LacZ reporter mice; immunofluorescence for osteoblast cilia localization; qRT-PCR and in situ hybridization of osteoblast and Wnt/β-catenin markers\",\n      \"journal\": \"Bone\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse model with multiple defined cellular phenotypes and marker analysis, direct cilia localization experiment\",\n      \"pmids\": [\"21911092\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Ectopic expression of EVC2 Weyers acrodental dysostosis missense variants (in the final exon of EVC2) in NIH 3T3 cells impairs Hedgehog signal transduction, consistent with a dominant-negative mechanism, whereas expression of a truncated EVC2 protein mimicking an Ellis-van Creveld loss-of-function mutation does not impair Hh signaling in this assay.\",\n      \"method\": \"Ectopic expression of murine EVC2 Weyers variants in NIH 3T3 cells; Hedgehog pathway reporter assays\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, functional cell-based assay but limited to overexpression without rescue controls\",\n      \"pmids\": [\"19810119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Evc is required for a spatially and temporally regulated Shh signaling response during molar development; absence of Evc causes progressive loss of Shh pathway response in a buccal-to-lingual direction and is associated with displaced Wnt pathway activity, establishing Evc as necessary for normal buccal-lingual axis patterning during tooth morphogenesis.\",\n      \"method\": \"Evc knockout mouse model; in situ hybridization for Shh target genes; LacZ reporter analysis for Wnt pathway; histological analysis of molar development\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse, multiple pathway markers, defined spatial and temporal phenotype, single lab\",\n      \"pmids\": [\"23315474\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WDR35 (IFT121), a retrograde intraflagellar transport protein, is specifically required for entry of EVC, EVC2, and Smoothened into the ciliary compartment. In Wdr35-/- cells all three proteins fail to localize to cilia, but this defect is not seen in Dync2h1-/- (retrograde motor mutant) cilia, establishing that IFT-A but not the retrograde IFT motor is required for EVC/EVC2/SMO ciliary targeting.\",\n      \"method\": \"Wdr35-/- and Dync2h1-/- mouse fibroblast cell lines; immunofluorescence for EVC, EVC2, and SMO cilia localization; Hedgehog pathway activity assays; expression of Wdr35 disease cDNAs for rescue\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple mutant cell lines compared, rescue by cDNA re-expression, multiple protein targets analyzed by direct imaging, mechanistic distinction between IFT-A and retrograde motor roles established\",\n      \"pmids\": [\"25908617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The EVC-EVC2 complex is subject to ubiquitination and SUMOylation of its cytosolic tails. Monoubiquitination of EVC-EVC2 cytosolic tails reduces protein levels. SUMOylation with SUMO3 enhances EVC-EVC2 accumulation at the EvC zone at the base of cilia, likely via increased binding to the EFCAB7-IQCE complex. A second EFCAB7-binding motif in EVC2's Weyers-deleted peptide was mapped, in addition to the previously characterized one. Known interactors EVC2, IQCE, EFCAB7 were confirmed, and USP7 (a deubiquitinase involved in Hh signaling) was identified as a new interactor, though ubiquitination of EVC-EVC2 was found to be independent of USP7 and USP48.\",\n      \"method\": \"Endogenous EVC protein interactome by mass spectrometry in control and Evc-null cells; ubiquitination assays; SUMO modification assays; immunofluorescence for EvC zone localization; domain mapping of EFCAB7-binding motifs\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteomic interactome screen plus direct modification assays and localization imaging, but single lab and some findings still require independent replication\",\n      \"pmids\": [\"37576597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Hypomorphic EVC missense mutations (p.Arg663Pro and splice variant c.1316-7A>G) produce proteins that retain partial ability to complex with EVC2, whereas the p.Arg622Ter nonsense mutation is a complete loss-of-function allele that cannot complex with EVC2, establishing that the degree of EVC-EVC2 complex formation correlates with clinical severity.\",\n      \"method\": \"Patient-derived fibroblasts; complementary DNA sequencing; immunoblot; immunofluorescence; Evc-/- mouse embryonic fibroblast complementation assays\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — patient fibroblasts with multiple orthogonal methods and mouse MEF complementation, single lab\",\n      \"pmids\": [\"32906221\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVC protein facilitates Shh signaling in notochord-derived nucleus pulposus cells; loss of EVC reduces Gli3 processing, impairs Shh pathway activity, and alters extracellular matrix organization toward fibrosis. TGF-β signaling suppresses EVC expression, indicating crosstalk between TGF-β and Hedgehog pathways at the level of EVC.\",\n      \"method\": \"Evc knockout mouse model; CRISPR-engineered human NP cells; proteome profiling; Gli3 processing western blot; ECM composition analysis; TGF-β treatment assays\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two model systems (mouse KO and CRISPR human cells), multiple pathway readouts, single lab\",\n      \"pmids\": [\"41550739\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVC and EVC2 are aberrantly overexpressed in a subset of AMLs (particularly those with ASXL1 mutations or t(8;21) translocation); functional loss of EVC/EVC2 impairs leukemia cell proliferation, promotes differentiation, and blocks AML progression in vivo. The leukemogenic role of EVC/EVC2 is mediated through MYC pathway activation, independent of their canonical role in Hedgehog signaling.\",\n      \"method\": \"shRNA/CRISPR loss-of-function in AML cell lines; in vivo AML transplantation models; ChIP-seq for AML1-ETO occupancy at EVC/EVC2 promoters; chromatin interaction analysis; MYC pathway gene expression analysis\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple functional assays (in vitro and in vivo) with mechanistic pathway placement, single lab, pathway independence from Hh needs further confirmation\",\n      \"pmids\": [\"41249566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Evc protein is expressed in prehypertrophic and hypertrophic chondrocytes of the epiphyseal growth plate in rat, as determined by immunohistochemistry and in situ hybridization, consistent with a function in chondrocyte differentiation during endochondral ossification.\",\n      \"method\": \"Immunohistochemistry with anti-Evc antibodies; in situ hybridization; RT-PCR on growth plate tissue\",\n      \"journal\": \"The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, localization only without direct functional consequence shown\",\n      \"pmids\": [\"15278943\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EVC encodes a transmembrane ciliary protein that localizes to a signaling microdomain (EvC zone) at the base of primary cilia, where it forms an obligate heterodimeric complex with EVC2; upon Hedgehog pathway activation, phosphorylated Smoothened recruits the EVC-EVC2 complex, which then promotes dissociation of Sufu from Gli3, facilitates Gli3 trafficking to cilia tips, and drives transcriptional activation of Hh target genes—a process regulated by ubiquitination (reducing complex stability) and SUMOylation (enhancing EvC zone accumulation via EFCAB7-IQCE tethering), with ciliary entry of the complex dependent on IFT-A machinery including WDR35.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EVC is a transmembrane ciliary protein that functions as an intracellular transducer of Hedgehog signaling, acting downstream of Smoothened but upstream of Gli3 processing to drive transcriptional activation of Hh target genes such as Ptch1 and Gli1 [#0]. It forms an obligate heterodimeric complex with EVC2, the two proteins binding directly and depending on each other for both ciliary localization and protein stability [#1, #3]. The complex localizes to a signaling microdomain at the base of the primary cilium, the EvC zone, where it is anchored by the EFCAB7-IQCE complex through binding of EFCAB7 to a C-terminal disordered region of EVC2; loss of this tethering mislocalizes EVC-EVC2 and impairs GLI2 activation [#4]. Upon Hh activation, phosphorylated Smoothened recruits EVC-EVC2 through its C-terminal tail in a cilium-dependent manner [#2], and the complex promotes Sufu/Gli3 dissociation and Gli3 trafficking to cilia tips [#3]; EVC-EVC2 also potentiates Hh output independently of Sufu [#2, #3]. Ciliary entry of EVC, EVC2, and Smoothened requires the IFT-A protein WDR35, distinct from the retrograde IFT motor [#8]. The complex is regulated post-translationally, with monoubiquitination of the cytosolic tails reducing protein levels and SUMOylation enhancing EvC zone accumulation via increased EFCAB7-IQCE binding [#9]. Through this Hh-transducing role, EVC governs chondrocyte proliferation and hypertrophy and osteoblast differentiation during endochondral ossification [#5], tooth buccal-lingual axis patterning [#7], and nucleus pulposus extracellular matrix homeostasis [#11]. Hypomorphic versus null EVC alleles differ in their ability to complex with EVC2, and the degree of complex formation correlates with the clinical severity of EVC-related skeletal dysplasia [#10]; dominant-negative EVC2 variants underlie Weyers acrodental dysostosis [#6]. EVC and EVC2 are also aberrantly overexpressed in subsets of AML, where they support leukemic proliferation through MYC pathway activation independent of Hedgehog signaling [#12].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established EVC as a bona fide intracellular Hedgehog pathway component by placing its action genetically between Smoothened and Gli3 processing, resolving where in the pathway it operates.\",\n      \"evidence\": \"Evc knockout mouse and Evc-/- cells with basal body immunofluorescence, Gli3 western blot, and qRT-PCR of Ihh targets\",\n      \"pmids\": [\"17660199\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular partners mediating transduction not yet identified\", \"Mechanism of basal body targeting unknown\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identified EVC2 as the direct, mutually co-dependent partner of EVC, defining the functional unit as a heterodimer rather than a solo protein.\",\n      \"evidence\": \"Yeast two-hybrid screen with EVC bait, co-immunoprecipitation, immunofluorescence, and subcellular fractionation\",\n      \"pmids\": [\"21356043\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and structure of the complex not determined\", \"Functional significance of nuclear full-length EVC2 unexplored\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Extended EVC's role beyond chondrocytes to osteoblasts and to both Pthrp-dependent and -independent Ihh functions, demonstrating cell-type-specific deployment within skeletal development.\",\n      \"evidence\": \"Evc-/- mice with Ptch1-LacZ reporter, osteoblast cilia immunofluorescence, and marker analysis\",\n      \"pmids\": [\"21911092\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Basis for differential requirement across perichondrial layers unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed that Hh activation drives phosphorylation-dependent binding of EVC-EVC2 to the Smoothened C-terminal tail and that the complex acts upstream of Sufu, mechanistically coupling Smo activation to downstream Gli activation.\",\n      \"evidence\": \"Co-IP of Smo with Evc/Evc2, constitutively active Smo, and Sufu-/- / Kif3a-/- cell lines with reporter assays\",\n      \"pmids\": [\"22986504\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the kinase generating the phospho-dependent binding site not defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined the downstream output of EVC-EVC2 as promoting Sufu/Gli3 dissociation and Gli3 ciliary tip recruitment, and showed a dominant Weyers Evc2 mutation mislocalizes the complex and reproduces these defects.\",\n      \"evidence\": \"Evc and Evc2 knockout mice, Co-IP, cilia tip immunofluorescence, Sufu/Gli3 dissociation western blot, and siRNA in Sufu-/- cells\",\n      \"pmids\": [\"23026747\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical action on the Sufu-Gli3 complex not reconstituted\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed how EVC-EVC2 is spatially confined to the EvC zone, identifying the EFCAB7-IQCE complex as the anchor that binds the Weyers-deleted region of EVC2 and is required for GLI2 activation.\",\n      \"evidence\": \"Co-IP with domain mapping, siRNA depletion of EFCAB7, and GLI2 activation assays\",\n      \"pmids\": [\"24582806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How EvC zone confinement mechanistically enables signaling not fully resolved\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified the trafficking route into cilia, showing IFT-A protein WDR35 — but not the retrograde motor — is required for ciliary entry of EVC, EVC2, and SMO.\",\n      \"evidence\": \"Wdr35-/- vs Dync2h1-/- fibroblasts, cilia localization imaging, pathway assays, and disease cDNA rescue\",\n      \"pmids\": [\"25908617\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct physical interaction between IFT-A and EVC cargo not demonstrated\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established post-translational control of the complex, showing monoubiquitination lowers EVC-EVC2 levels while SUMOylation boosts EvC zone accumulation, and mapped a second EFCAB7-binding motif plus a USP7 interaction.\",\n      \"evidence\": \"Endogenous EVC interactome mass spectrometry in control vs Evc-null cells, ubiquitination/SUMO assays, and localization imaging\",\n      \"pmids\": [\"37576597\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ubiquitin ligase responsible not identified\", \"Functional role of the USP7 interaction unresolved\", \"Single lab, some findings await replication\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected genotype to phenotype by showing that EVC allele severity tracks with residual ability to complex with EVC2, providing a molecular basis for clinical variability.\",\n      \"evidence\": \"Patient fibroblasts, immunoblot/immunofluorescence, and Evc-/- MEF complementation assays\",\n      \"pmids\": [\"32906221\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Quantitative threshold of complex formation needed for function not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Distinguished the disease mechanisms of two EVC2-linked dysplasias, showing Weyers missense variants act dominant-negatively on Hh signaling while a truncating Ellis-van Creveld-type allele does not in the assay.\",\n      \"evidence\": \"Ectopic expression of murine EVC2 variants in NIH 3T3 cells with Hh reporter assays\",\n      \"pmids\": [\"19810119\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Overexpression assay without rescue controls\", \"Endogenous-level confirmation lacking\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showed EVC is required for spatiotemporal Shh response and buccal-lingual axis patterning in tooth development, broadening its developmental remit.\",\n      \"evidence\": \"Evc knockout mice with in situ hybridization, Wnt LacZ reporter, and histology of molar development\",\n      \"pmids\": [\"23315474\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking EVC loss to displaced Wnt activity not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended EVC function to nucleus pulposus ECM homeostasis and uncovered TGF-beta suppression of EVC, identifying pathway crosstalk affecting Shh output and fibrosis.\",\n      \"evidence\": \"Evc knockout mice and CRISPR human NP cells with Gli3 western blot, proteome profiling, and TGF-beta treatment\",\n      \"pmids\": [\"41550739\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of TGF-beta-mediated EVC repression unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealed a non-canonical oncogenic role in AML where EVC/EVC2 overexpression supports leukemic proliferation via MYC, independent of Hedgehog signaling.\",\n      \"evidence\": \"shRNA/CRISPR loss-of-function in AML lines, in vivo transplantation, AML1-ETO ChIP-seq, and MYC pathway expression analysis\",\n      \"pmids\": [\"41249566\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link between EVC and MYC unresolved\", \"Hh-independence requires further confirmation\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the EVC-EVC2 complex biochemically catalyzes Sufu/Gli3 dissociation, the structure of the assembled complex, and the identity of the enzymes controlling its ubiquitination remain unresolved.\",\n      \"evidence\": \"Not addressed by current timeline\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No reconstituted biochemical mechanism for Sufu/Gli3 dissociation\", \"No structural model of the EVC-EVC2 complex\", \"Ubiquitin ligase identity unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2, 3, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1, 3, 4]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [5, 7, 11]}\n    ],\n    \"complexes\": [\"EVC-EVC2 complex\", \"EvC zone (EVC-EVC2-EFCAB7-IQCE)\"],\n    \"partners\": [\"EVC2\", \"SMO\", \"EFCAB7\", \"IQCE\", \"USP7\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":9,"faith_total":9,"faith_pct":100.0}}