EVC participates in a ciliary membrane complex with EVC2 and positively supports Hedgehog signaling downstream of Smoothened. The mechanistic originals inspected here principally assay mouse proteins. Human HEK293 or HEK293T expression hosts do not convert those reagents into human EVC. Current human database models and antibody imaging provide additional, differently scoped evidence.
Blair et al., 2011, PMID:21356043 was read in the actual main-text XML, including Results, Methods and captions. Coexpression/localization and topology experiments use complete mouse Evc/Evc2 constructs and native mouse cells. The intracellular Evc C terminus and restricted ciliary membrane distribution support a more precise location than general plasma membrane or cilium. The interaction Results describe Evc 463–1005, whereas Methods describe 463–991; that inconsistency is retained rather than silently corrected. Some full-length expression/co-immunoprecipitation experiments were limited by solubility. Evc2 nuclear observations are not assigned to Evc. Figure pixels and supplementary phylogeny were not inspected.
Pusapati et al., 2014, PMID:24582806 was read through the original BioC body: Results, Discussion, operative main Methods and main captions. Native NIH/3T3 complex purification, co-immunoprecipitation and biochemical fractionation distinguish EVC–EVC2 and EFCAB7–IQCE subcomplexes. All EvC constructs are mouse, including those expressed in human HEK293T. EFCAB7 supplies the demonstrated bridging interactions; EVC is not thereby assigned EFCAB7 adaptor activity. Loss of EFCAB7 allows EVC/EVC2 to spread along ciliary membrane. A scaffold mechanism for the EVC complex is discussed as a possibility, with no detected downstream SUFU/GLI association in the reported experiments. A fixed obligatory four-subunit assembly or a measured EVC scaffold molecular function is not inferred. Figure pixels and supplemental experiments were not read.
The current human Reactome assembly event R-HSA-5633051 contains EVC2:EVC complex 5633041. Its EVC component 5633035 explicitly identifies P57679, human 9606, residues 1–992 and its own ciliary-membrane compartment. The localization is therefore attached to the protein itself, not borrowed solely from the event. This is a human pathway model informed by ortholog experiments; it is not another human localization experiment or evidence that EVC catalyzes a neighboring reaction.
Ruiz-Perez et al., 2007, PMID:17660199 was read in the publisher-provided author preview, captured lines 331–987. The native mouse ciliary-base staining used an antibody raised against mouse CAB76567 residues 459–999, with knockout loss of staining. The authors describe distal basal-body/transition-zone positioning from light microscopy; they do not establish an ultrastructural position within the basal-body microtubule array. Main Methods, growth-plate Results, pathway readouts and purmorphamine responses were read, but the later differentiation section extends beyond the recovered passage. No original figure pixels or supplements were inspected.
The current human HPA EVC page independently lists approved basal body and nucleoplasm as main locations, with primary cilium and cytosol as additional locations. HPA016046 reports basal body in ASC52telo, serum-starved hTERT-RPE1 and RPTEC/TERT1. Two selected ciliated-cell images were actually inspected: RPE1 shows green puncta at the bases of red ciliary profiles; ASC52 signal is weaker and also diffuse. The downloaded composites show blue/red/green channels, without an independently visible yellow basal-body-marker channel. This is positive localization evidence with spatial-resolution and antibody limits, not quantitative colocalization, a nuclear mechanism, or evidence that adjacent ciliary-membrane localization excludes the basal body.
The HPA016046 antigen exactly matches residues 78–180 of current P57679 and all three captured NP products. It therefore cannot establish which product contributes the signal. No localization or function is automatically assigned to the distinct 991-aa or 535-aa RefSeq products. The imported UniProt record contains one displayed 992-aa sequence and no alternative-product comment; external records remain separate and no artificial UniProt isoform identifiers are created.
The actual PMID:17660199 mouse experiments connect Evc loss to shorter endochondral bones, altered growth-plate zones and reduced spatial Gli1/Ptch1 responses despite retained Ihh expression. Impaired purmorphamine responses place the positive contribution downstream of Smoothened activation. This supports signaling participation rather than merely a necessity inference from skeletal abnormality. Bone growth and cartilage progression remain downstream developmental contexts; no bone/cartilage-matrix synthetic activity is assigned to EVC.
PMID:15278943(https://doi.org/10.1002/ar.a.20059), the proximate rat cartilage-development source, was verified only through metadata and its complete abstract. It reports Evc transcript/protein distribution in prehypertrophic and hypertrophic growth-plate chondrocytes. Its operative original body remains unread. Expression alone is insufficient to establish developmental participation. Retention of the existing cartilage-development context instead uses the independent mouse signaling and growth-plate experiments above, with the rat-source limitation explicit. No wrong-gene or curator-error inference follows from the access gap.
Actual PANTHER PTHR16795 tree, alignment and current PAINT IBD records were examined. Human P57679 leaf PTN002497663 descends from PTN000423571; its ungapped aligned sequence exactly matches 992-aa EVC. The mouse leaf also exactly matches the current 1,005-residue sequence. The rat leaf is 990 aa whereas the current G3V706 record is 1,003 aa; those sequences are not equated. Rat identity and the current IEP source are verified separately, and this version/product discrepancy does not establish functional divergence or invalidate the human PAINT placement. Positive ciliary-membrane and membrane-complex IBDs lie at the source node, grounded by the experimentally characterized mouse Evc/Evc2 genes. No target-path NOT was found. The shared ancestral node supports conserved complex biology; donor count is not a quality measure.
InterPro IPR026501 is an actual target family match and maps Smoothened signaling. Pathway participation and positive regulation are both supported by the operative Evc-specific experiments. The regulation term is not an ancestry-equivalent substitute for the pathway term; neither asserts enzyme activity. Current UniProt subcellular mappings SL-0039, SL-0066 and SL-0305 correctly map plasma membrane, cilium and ciliary membrane. The human participant and original topology evidence justify refinement of the broader locations.
The human disease-discovery paper, PMID:10700184, is verified against PubMed/Europe PMC and the publisher preview. Its accessible abstract links EVC variants to skeletal dysplasia and cardiac malformations. The full body, Methods and figures remain unavailable; the nominal publisher PDF response was HTML. The broad skeletal-development annotation is retained as a downstream non-core context. The muscle-organ development assertion remains UNDECIDED because the accessible abstract does not resolve its specific basis; this is not a claim that the original curators were wrong.
The positive human ciliary localization is retained and refined to ciliary membrane despite the mouse-construct and native-mouse-cell scope of PMID:21356043; those experiments do not contradict the conserved location. A probable, unreported source query asks UniProt to check the human IDA evidence basis or identify ortholog-transfer provenance. No direct human localization experiment, corrected evidence code or replacement gene accession is invented.
The single core combines participation in Smoothened signaling and positive regulation of its output with ciliary-membrane and basal-body-region localization and plasma-membrane-complex membership. The operative mouse Evc-null agonist and pathway-readout experiments in PMID:17660199(https://doi.org/10.1242/dev.007542) establish a positive signaling contribution. The localization/topology and complex experiments in PMID:21356043(https://doi.org/10.1186/1741-7007-9-14) and PMID:24582806(https://doi.org/10.1016/j.devcel.2014.01.021) place that contribution in an EVC2-associated proximal ciliary membrane complex. Human Reactome participant 5633035 supplies a concordant native-protein location model, not an independent human assay.
PMID:24582806 Discussion, specifically the paragraph beginning with the need for further mechanistic research, distinguishes a possible scaffold mechanism from demonstrated activity. The study did not detect interactions with downstream PKA, SUFU or GLI proteins under its conditions. Its EFCAB7 adaptor model is supported by EFCAB7-specific interaction mapping and recruitment experiments; that activity is not assigned to EVC. No molecular-function term is supplied for EVC because the examined experiments do not establish its individual biochemical step. This is a bounded biological knowledge gap, not a claim that the protein lacks function or that no future work can establish a scaffold mechanism.
Basal-body association is represented in the same signaling core alongside ciliary membrane, with the spatial-resolution limits described above. No separate nuclear, basal-body-assembly or developmental core is inferred. No fixed complex stoichiometry or functional equivalence of the 992-, 991- and 535-residue human products is assumed.
The unchanged UniProt P57679 record, sections CC FUNCTION and CC SUBCELLULAR LOCATION, supplies curated human signaling and location statements. These statements complement the mouse experiments; they are not new human assays. GO_REF:0000052 defines an immunofluorescence curation method and is retained as provenance, not as target-specific localization evidence.
For the fragment discrepancy, see the complete 2011 original, Results “Evc interacts with Evc2” and Methods “Plasmids”. The interaction Results describe the mouse Evc fragment as residues 463–1005, whereas the co-immunoprecipitation construct in Methods is 463–991. These are the paper’s two descriptions, not an inferred correction or human-product coordinate mapping. The same original’s Results on cytoplasmic/nuclear localization, Figure 7 and Discussion report full-length Evc only in the cytoplasmic fraction of mouse MEFs, with Evc2 in both fractions. Null-cell and fraction-marker controls support that result; the authors allow an Evc nuclear pool below detection. It therefore informs, but does not negate, the independent human HPA nucleoplasmic observation.
The HPA evidence is publicly accessible at the EVC subcellular page and HPA016046 antibody page. The inspected composites are ASC52telo field 34 and serum-starved hTERT-RPE1 field 19. These blue/red/green images show ciliary-base puncta most clearly in RPE1; a separate yellow basal-body marker channel is not displayed. The approved antibody classification and cell-specific calls do not establish ultrastructural incorporation or a nuclear function. The antigen is shared by all three versioned RefSeq products below.
| Public protein record | Sequence length | Scope |
|---|---|---|
| NP_714928.1 | 992 residues | Sequence matches UniProt P57679. |
| NP_001293019.1 | 991 residues | Distinct RefSeq product; no functional equivalence assumed. |
| NP_001293021.1 | 535 residues | Distinct RefSeq product; no functional equivalence assumed. |
These lengths come from the versioned NCBI protein sequences, not from a UniProt alternative-products comment. All three contain the HPA016046 antigen at residues 78–180. Metadata and a shared antibody epitope do not establish expression, localization or activity of every product.
The 2014 original, Discussion, reports signal-dependent association of the EVC–EVC2 complex with SMO. It also leaves the molecular link to downstream effectors unresolved. This is a useful boundary for the MF_DARK gap: a receptor-associated signaling complex is supported, while a specific EVC adaptor/scaffold activity or isolated EVC–SMO binary interaction is not established by those experiments. Experimental Procedures, Plasmids, identifies the mouse EvC constructs; HEK293T is an expression host.
The campaign curation instructions retain the user’s no-new-publication-quotations rule. Evidence here uses verified reference identifiers and public section, image and accession locators; no verbatim-substring validation is claimed for reference-only support. The same instructions retain supported generic binding as non-core when no specific replacement is established; EVC has no generic-binding row requiring a decision in this revision. No new annotation is proposed in this bounded revision because the supported claims already have source rows. This is a scope decision, not a project-wide prohibition on NEW annotations.
