Affinage

EVC2

Limbin · UniProt Q86UK5

Length
1308 aa
Mass
147.9 kDa
Annotated
2026-06-09
39 papers in source corpus 11 papers cited in narrative 11 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

EVC2 is a ciliary transmembrane protein that transduces Hedgehog (Hh) signaling at the EvC zone, a specialized microdomain at the base of primary cilia (PMID:22981989). EVC2 forms a constitutive, mutually dependent heterodimer with EVC: the two proteins require each other for ciliary and basal-body localization and for maintaining each other's protein levels (PMID:23026747, PMID:21356043). Upon Hh stimulation, the EVC-EVC2 complex binds Smoothened in a manner that depends on phosphorylation of the Smo C-terminal tail and on intact primary cilia, and this binding occurs specifically within the EvC zone (PMID:22981989, PMID:22986504). EVC2 acts downstream of Smo to promote Gli transcription factor activation, driving Gli3 recruitment to cilia tips and Sufu/Gli3 dissociation, with additional positive activity downstream of Sufu (PMID:22986504, PMID:23026747). Anchoring at the EvC zone requires the EFCAB7-IQCE complex, which binds a C-terminal disordered region of EVC2 deleted in Weyers acrodental dysostosis; loss of this anchoring mislocalizes the complex within cilia and impairs Gli activation (PMID:24582806, PMID:37576597). Weyers-associated EVC2 C-terminal mutations act as dominant-negative alleles that retain ciliary localization but are displaced from the EvC zone (PMID:22981989, PMID:19810119). In vivo, Evc2 is required in chondrocytes and dental mesenchyme for skeletal and tooth development, in part by restraining perichondrial FGF18 signaling in the growth plate (PMID:26219237, PMID:28027321, PMID:28081373). Independently of its Hedgehog role, EVC2 (with EVC) is aberrantly overexpressed in a subset of AML, where it promotes leukemia progression through MYC pathway activation (PMID:41249566).

Mechanistic history

Synthesis pass · year-by-year structured walk · 9 steps
  1. 2009 Medium

    Established that the EVC2 C-terminus is functionally critical, showing Weyers missense mutations impair Hh signaling through a dominant-negative rather than simple loss-of-function mechanism.

    Evidence Ectopic expression of murine Weyers and Ellis-van Creveld-mimicking EVC2 variants in NIH 3T3 cells with Hh reporter assays

    PMID:19810119

    Open questions at the time
    • Single lab, not independently replicated
    • Did not define the molecular partner or microdomain whose binding is disrupted
    • No structural basis for the dominant-negative effect
  2. 2011 High

    Defined the core physical interaction and topology, showing EVC2 and EVC directly bind and co-dependently localize to the basal body and primary cilium.

    Evidence Yeast two-hybrid, Co-IP, confocal immunofluorescence, subcellular fractionation, and Hh reporter assays in Evc2-null cells

    PMID:21356043

    Open questions at the time
    • Functional significance of nuclear full-length EVC2 not established
    • Did not place EVC-EVC2 relative to Smo or Sufu in the pathway
    • Mechanism of co-dependent stabilization unresolved
  3. 2012 High

    Resolved where in the Hh pathway EVC-EVC2 acts, showing it binds phospho-Smo within the EvC zone in a cilium-dependent manner and promotes Gli activation by driving Sufu/Gli3 dissociation upstream and downstream of Sufu.

    Evidence Reciprocal Co-IP, phosphorylation-dependent Smo-binding assays, dominant-negative localization mutants, epistasis in Sufu-/-/Kif3a-/- cells, and Evc2-/- / Evc2Δ43 knock-in mice across three independent studies

    PMID:22981989 PMID:22986504 PMID:23026747

    Open questions at the time
    • Molecular mechanism by which EVC-EVC2 triggers Sufu/Gli3 dissociation not defined
    • Structure of the Smo-EVC2 interface unknown
    • How the EvC zone is physically distinguished from the rest of the ciliary base unresolved
  4. 2014 High

    Identified the anchoring mechanism, showing EFCAB7-IQCE tethers EVC-EVC2 to the EvC zone via the same C-terminal region deleted in Weyers syndrome.

    Evidence Reciprocal Co-IP, domain mapping, siRNA depletion with localization and Hh reporter phenotypes

    PMID:24582806

    Open questions at the time
    • How EFCAB7-IQCE itself is positioned at the EvC zone not established
    • Stoichiometry of the EVC-EVC2-EFCAB7-IQCE assembly unknown
  5. 2015 High

    Demonstrated the in vivo tissue requirements, showing Evc2 loss eliminates ciliary EVC/EVC2, reduces Hh signaling, and causes skeletal and oral defects with cartilage and neural-crest specificity.

    Evidence Global and tissue-specific conditional KO mice with ciliary localization imaging, Hh readouts, and skeletal phenotyping

    PMID:26219237

    Open questions at the time
    • Osteoblast-specific deletion phenotype absent, leaving osteoblast role undefined
    • Cell-autonomous vs non-autonomous contributions not fully separated
  6. 2016 High

    Connected EVC2 loss to a downstream pathogenic mechanism, showing elevated perichondrial FGF18 contributes to limb dwarfism and is partially rescued genetically.

    Evidence Evc2 mutant mice, FGF signaling assays, and Fgf18+/- x Evc2-/- double-mutant genetic rescue

    PMID:28027321

    Open questions at the time
    • Mechanism linking reduced Hh signaling to elevated Fgf18 not defined
    • Partial rescue implies additional pathogenic pathways
  7. 2017 Medium

    Localized the dental requirement, showing Evc2 acts in dental mesenchyme for odontoblast differentiation, with ameloblast defects arising secondarily.

    Evidence Dental mesenchyme-specific conditional KO mice with marker immunohistochemistry and stem-cell analysis

    PMID:28081373

    Open questions at the time
    • Single lab
    • Signaling pathway mediating the mesenchymal effect not resolved
  8. 2023 Medium

    Revealed post-translational regulation of complex abundance and localization, showing monoubiquitination lowers EVC-EVC2 levels while SUMO3 modification enhances EvC zone accumulation.

    Evidence Endogenous interactome MS, ubiquitination/SUMOylation assays, localization imaging, and EFCAB7-binding motif mapping

    PMID:37576597

    Open questions at the time
    • The deubiquitinase responsible was not identified (USP7/USP48 excluded)
    • The ubiquitin ligase and SUMO machinery acting on EVC-EVC2 unknown
    • Functional role of USP7 as an interactor undefined
  9. 2025 Medium

    Uncovered a Hedgehog-independent oncogenic role, showing EVC/EVC2 overexpression drives a subset of AML through MYC pathway activation.

    Evidence Knockdown/KO in AML cell lines, in vivo progression assays, MYC readouts, and chromatin occupancy/interaction analysis

    PMID:41249566

    Open questions at the time
    • Single study, not replicated
    • Mechanism by which EVC/EVC2 activates MYC is undefined
    • Whether ciliary localization is involved in the AML role unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the EVC-EVC2 complex mechanistically couples phospho-Smo binding at the EvC zone to Sufu/Gli3 dissociation, and how this ciliary protein contributes to Hh-independent MYC activation in AML, remain unresolved.
  • No structural model of the EVC-EVC2-Smo interface
  • Biochemical mechanism of Sufu/Gli3 release not established
  • MYC-activating mechanism in AML undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060089 molecular transducer activity 3 GO:0060090 molecular adaptor activity 2
Localization
GO:0005929 cilium 4 GO:0005815 microtubule organizing center 1 GO:0005886 plasma membrane 1
Pathway
R-HSA-1266738 Developmental Biology 3 R-HSA-162582 Signal Transduction 3 R-HSA-1643685 Disease 1
Partners
EFCAB7EVCIQCESMOUSP7
Complex memberships
EVC-EVC2 complexEvC zone (EVC-EVC2-EFCAB7-IQCE)

Evidence

Reading pass · 11 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2012 Hedgehog agonists promote physical association between Smoothened (Smo) and EVC2; this Smo-EVC2 complex forms specifically within the EvC zone, a distinct sub-compartment at the base of primary cilia. EVC2 mutants that localize to cilia but are displaced from the EvC zone act as dominant inhibitors of Hh signaling, and loss of EVC2 function blocks Hh signaling between Smo and the downstream regulators PKA and Suppressor of Fused, preventing Gli transcription factor activation. Co-immunoprecipitation, dominant-negative localization mutant analysis, epistasis with PKA/Sufu/Gli in cell-based assays, live imaging of ciliary localization Developmental cell High 22981989
2012 Hh stimulation induces binding of EVC/EVC2 to Smo in a manner that depends on phosphorylation of the Smo C-terminal intracellular tail; this binding is abolished in Kif3a-/- cilium-deficient cells, establishing that the interaction requires primary cilia. EVC/EVC2 acts upstream of Sufu to promote Gli activation, as EVC/EVC2 is dispensable for constitutive Gli activity in Sufu-/- cells. Co-immunoprecipitation, phosphorylation-dependent binding assay using Smo mutants, epistasis in Sufu-/- and Kif3a-/- cells Cell research High 22986504
2012 EVC and EVC2 are mutually required for their localization to primary cilia and for maintaining normal protein levels of each other; in Evc2-deficient chondrocytes, Smo translocation to cilia is normal after Hh activation, but Gli3 recruitment to cilia tips is reduced and Sufu/Gli3 dissociation is impaired. Smo co-precipitates with the EVC/EVC2 complex. Expression of the Weyers-associated Evc2Δ43 mutant causes mislocalization of EVC/EVC2 within cilia and reproduces the Gli3 molecular defects. EVC silencing in Sufu-/- cells attenuates Hh pathway output, indicating EVC/EVC2 also promotes signaling downstream of Sufu. Mouse genetic models (Evc2-/-, Evc2Δ43 knock-in), Co-immunoprecipitation, immunofluorescence of ciliary Gli3/Sufu localization, Evc/Evc2 double mutant analysis Human molecular genetics High 23026747
2011 EVC2 and EVC directly interact (identified by yeast two-hybrid with EVC as bait, confirmed by co-immunoprecipitation). EVC2 and EVC co-localize at the basal body and on primary cilia, and their localization is co-dependent: ciliary/basal body localization requires co-transfection of both constructs. EVC2 is a transmembrane protein with an extracellular N-terminus and intracellular C-terminus, while EVC is fully intracellular. Full-length EVC2 (but not EVC) is also detected in the nucleus by Western blotting of nuclear fractions. EVC2 is a positive regulator of Hh pathway activation in response to Smo agonist purmorphamine. Yeast two-hybrid, co-immunoprecipitation, immunofluorescence/confocal microscopy, subcellular fractionation + Western blot, Hh reporter assay in Evc2-null cells BMC biology High 21356043
2014 EFCAB7 and IQCE form a complex that anchors the EVC-EVC2 complex to the EvC zone at the base of primary cilia. EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. EFCAB7 depletion causes mislocalization of EVC-EVC2 within cilia (phenocopying the Weyers cellular phenotype) and impairs GLI2 activation. Co-immunoprecipitation, immunofluorescence localization, siRNA knockdown, Hh pathway reporter assays, evolutionary/bioinformatic analysis of EVC2 C-terminal region Developmental cell High 24582806
2009 Expression of Weyers acrodental dysostosis EVC2 exon 22 missense mutations (but not a truncation mimicking Ellis-van Creveld) impairs Hedgehog signal transduction in NIH 3T3 cells, demonstrating a dominant-negative mechanism for Weyers mutations at the EVC2 C-terminus. Ectopic expression of murine Weyers EVC2 variants and truncation mutants in NIH 3T3 fibroblasts, Hh pathway reporter assays Human mutation Medium 19810119
2015 Global and conditional knockout of Evc2/Limbin in mice eliminates ciliary localization of both EVC2 and EVC proteins, reduces Hedgehog signaling activity, and causes skeletal and oral defects. Cartilage-specific deletion recapitulates skeletal defects; neural crest-specific deletion causes incisor growth defects. Osteoblast-specific deletion does not cause overt skeletal changes. Conditional KO mouse generation (Cre-lox), immunofluorescence for ciliary localization, Hh signaling readouts (target gene expression), skeletal/histological phenotyping Genesis High 26219237
2016 In Evc2 mutant mouse growth plates, FGF signaling is elevated in addition to compromised Hh-PTHrP feedback. Elevated Fgf18 expression from the perichondrium upon Evc2 inactivation is a critical pathogenic contributor to limb dwarfism, as heterozygous deletion of Fgf18 in Evc2 mutants partially rescues the limb dwarfism phenotype. Evc2 mutant mouse analysis, in vivo and in vitro FGF signaling assays, genetic rescue (Fgf18+/- × Evc2-/- double mutant mice), gene expression analysis PLoS genetics High 28027321
2017 Dental mesenchymal-specific deletion of Evc2 phenocopies the tooth abnormalities (enamel hypoplasia, delayed ameloblast differentiation) seen in global Evc2 mutants, establishing that Evc2 function in dental mesenchyme is required for normal odontoblast differentiation and dental mesenchymal stem cell homeostasis. Delayed odontoblast differentiation secondarily impairs ameloblast differentiation. Conditional KO mice (dental mesenchyme-specific Cre), histology, ameloblast/odontoblast marker immunohistochemistry, stem cell marker analysis Journal of dental research Medium 28081373
2023 The EVC-EVC2 complex is subject to ubiquitination and SUMOylation of the cytosolic tails. Monoubiquitination of EVC-EVC2 cytosolic tails reduces protein levels. SUMOylation with SUMO3 enhances EVC-EVC2 accumulation at the EvC zone, possibly via increased binding to the EFCAB7-IQCE complex. EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide. Proteomic screen of EVC interactome confirmed EVC2, IQCE, EFCAB7 as main interactors and identified USP7 as a novel interactor; however, USP7 (and USP48) are not responsible for EVC-EVC2 deubiquitination. Endogenous protein interactome (proteomics/MS), ubiquitination/SUMOylation assays, ciliary localization imaging, domain mapping of EFCAB7-binding motifs in EVC2 Frontiers in cell and developmental biology Medium 37576597
2025 EVC and EVC2 are aberrantly overexpressed in a subset of AML cells (particularly those with ASXL1 mutations or t(8;21)). Loss of EVC/EVC2 impairs leukemia cell proliferation, promotes differentiation, and blocks AML progression in vivo. The leukemogenic role is mediated through MYC pathway activation, independent of Hedgehog signaling. Elevated EVC/EVC2 expression is associated with gained AML1-ETO occupancy or enhanced chromatin interactions at EVC/EVC2 promoter regions. Functional knockdown/KO studies in AML cell lines, in vivo AML progression assays, MYC pathway readouts, chromatin interaction/occupancy analysis (ChIP/Hi-C type) Leukemia Medium 41249566

Source papers

Stage 0 corpus · 39 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2012 A Smoothened-Evc2 complex transduces the Hedgehog signal at primary cilia. Developmental cell 150 22981989
2012 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. Human molecular genetics 94 23026747
2012 Smoothened transduces Hedgehog signal by forming a complex with Evc/Evc2. Cell research 86 22986504
2006 Sequencing EVC and EVC2 identifies mutations in two-thirds of Ellis-van Creveld syndrome patients. Human genetics 80 17024374
2002 A new gene, EVC2, is mutated in Ellis-van Creveld syndrome. Molecular genetics and metabolism 80 12468274
2014 EFCAB7 and IQCE regulate hedgehog signaling by tethering the EVC-EVC2 complex to the base of primary cilia. Developmental cell 77 24582806
2011 Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus. BMC biology 77 21356043
2012 Novel and recurrent EVC and EVC2 mutations in Ellis-van Creveld syndrome and Weyers acrofacial dyostosis. European journal of medical genetics 59 23220543
2009 Widening the mutation spectrum of EVC and EVC2: ectopic expression of Weyer variants in NIH 3T3 fibroblasts disrupts Hedgehog signaling. Human mutation 52 19810119
2006 A novel heterozygous deletion in the EVC2 gene causes Weyers acrofacial dysostosis. Human genetics 40 16404586
2014 Deletion in the EVC2 gene causes chondrodysplastic dwarfism in Tyrolean Grey cattle. PloS one 28 24733244
2015 Generation of Evc2/Limbin global and conditional KO mice and its roles during mineralized tissue formation. Genesis (New York, N.Y. : 2000) 27 26219237
2011 Two novel heterozygous mutations of EVC2 cause a mild phenotype of Ellis-van Creveld syndrome in a Chinese family. American journal of medical genetics. Part A 20 21815252
2017 Ellis-van Creveld syndrome and profound deafness resulted by sequence variants in the EVC/EVC2 and TMC1 genes. Journal of genetics 19 29321360
2016 Elevated Fibroblast Growth Factor Signaling Is Critical for the Pathogenesis of the Dwarfism in Evc2/Limbin Mutant Mice. PLoS genetics 19 28027321
2017 Loss of Function of Evc2 in Dental Mesenchyme Leads to Hypomorphic Enamel. Journal of dental research 18 28081373
2017 The Role of Ellis-Van Creveld 2(EVC2) in Mice During Cranial Bone Development. Anatomical record (Hoboken, N.J. : 2007) 16 28950429
2018 A Ciliary Protein EVC2/LIMBIN Plays a Critical Role in the Skull Base for Mid-Facial Development. Frontiers in physiology 14 30410447
2016 Truncation and microdeletion of EVC/EVC2 with missense mutation of EFCAB7 in Ellis-van Creveld syndrome. Congenital anomalies 12 26748586
2016 Expression of Evc2 in craniofacial tissues and craniofacial bone defects in Evc2 knockout mouse. Archives of oral biology 11 27164562
1980 Change of H-2 antigens' expression on mouse leukaemia LBN/a-2 and LBN/b-3 cells in the course of serial transplantation. Journal of immunogenetics 9 7373065
2023 Establishing an objective clinical spectrum, genotype-phenotype correlations, and CRMP1 as a modifier in the Ellis-van Creveld syndrome: The first systematic review of EVC- and EVC2-associated conditions. Genetics in medicine open 8 39669252
2016 Novel homozygous mutations in the EVC and EVC2 genes in two consanguineous families segregating autosomal recessive Ellis-van Creveld syndrome. Clinical dysmorphology 8 26580685
2023 EVC-EVC2 complex stability and ciliary targeting are regulated by modification with ubiquitin and SUMO. Frontiers in cell and developmental biology 7 37576597
2012 Identification of one novel mutation in the EVC2 gene in a Chinese family with Ellis-van Creveld syndrome. Gene 7 23026208
2019 Novel deleterious mutation in MYO7A, TH and EVC2 in two Pakistani brothers with familial deafness. Pakistan journal of medical sciences 6 30881389
2022 Dental Anomalies in Ciliopathies: Lessons from Patients with BBS2, BBS7, and EVC2 Mutations. Genes 5 36672825
2021 Ellis-van Creveld syndrome novel pathogenic variant in the EVC2 gene a patient from Turkey. Clinical case reports 4 33936625
2026 Neural crest-specific disruption of Evc2 provides an animal model to study the temporomandibular joint development and homeostasis in response to jaw loading. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 3 41051362
1976 DNA polymerases of murine LBN/b leukemic cells. Acta biochimica Polonica 3 59500
2023 Identification of Compound Heterozygous EVC2 Gene Variants in Two Mexican Families with Ellis-van Creveld Syndrome. Genes 2 37107645
2023 Prenatal whole exome sequencing identified two rare compound heterozygous variants in EVC2 causing Ellis-van Creveld syndrome. Molecular genetics & genomic medicine 2 37485807
2023 The oligodontia phenotype in a X-linked hypohidrotic ectodermal dysplasia patient with a novel EVC2 variant. Heliyon 1 38163170
2022 Novel large deletion involving EVC and EVC2 in Ellis-van Creveld syndrome. Human genome variation 1 35581188
2026 A novel EVC2 splice-site variant expands the mutational and phenotypic spectrum of Weyers acrofacial dysostosis. BMC medical genomics 0 41664038
2025 Case Report: A novel compound heterozygosity of the EVC2 gene identified in a Chinese pedigree with congenital heart defect. Frontiers in pediatrics 0 40726901
2025 Oncogenic activation of EVC/EVC2 defines a therapeutically targetable subset of acute myeloid leukemia. Leukemia 0 41249566
2025 Identification of a Novel EVC2 Variant in a Family with Non-Syndromic Tooth Agenesis and Its Potential Functional Implications. Genes 0 41300740
2005 [From gene to disease; EVC, EVC2, and Ellis-van Creveld syndrome]. Nederlands tijdschrift voor geneeskunde 0 15884406

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