{"gene":"EFCAB7","run_date":"2026-04-28T17:46:03","timeline":{"discoveries":[{"year":2014,"finding":"EFCAB7 forms a complex with IQCE that anchors the EVC-EVC2 complex in a signaling microdomain at the base of primary cilia (the EvC zone). EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of the transcription factor GLI2, phenocopying the Weyers cellular phenotype.","method":"Co-immunoprecipitation, siRNA knockdown, fluorescence microscopy/live imaging of ciliary localization, GLI2 reporter assays, evolutionary/bioinformatic analysis","journal":"Developmental Cell","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, direct binding mapped to specific domain, KD with defined ciliary mislocalization phenotype and downstream signaling readout; replicated in part by subsequent studies","pmids":["24582806"],"is_preprint":false},{"year":2023,"finding":"SUMO3 modification of EVC-EVC2 cytosolic tails enhances their 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; only one was previously characterized, and the second was mapped in this study. Monoubiquitination of EVC-EVC2 cytosolic tails reduces their protein levels. Proteomic screen of the EVC interactome confirmed EVC2, IQCE, and EFCAB7 as core interactors.","method":"Endogenous protein interactome proteomics (AP-MS) in control and Evc-null cells, SUMO and ubiquitin modification assays, confocal microscopy of ciliary localization, domain mapping","journal":"Frontiers in Cell and Developmental Biology","confidence":"Medium","confidence_rationale":"Tier 2 — AP-MS interactome plus localization and modification assays from a single lab; moderate evidence for EFCAB7-binding motif mapping","pmids":["37576597"],"is_preprint":false},{"year":2016,"finding":"A heterozygous missense variant in EFCAB7 (c.1171T>C; p.Y391H) was identified in an Ellis-van Creveld patient also carrying an EVC truncation mutation, suggesting EFCAB7 acts as a modifier of EVC-related phenotypes through its interaction with EVC.","method":"Sanger sequencing, SNP array, mRNA expression analysis in patient samples","journal":"Congenital Anomalies","confidence":"Low","confidence_rationale":"Tier 3 — genetic/expression data from patient samples, no direct functional reconstitution","pmids":["26748586"],"is_preprint":false},{"year":2018,"finding":"A novel missense mutation in EFCAB7 was shown to alter the expression levels and interaction of Ellis-van Creveld ciliary proteins (EVC/EVC2/IQCE), supporting EFCAB7's role in stabilizing the EvC ciliary complex.","method":"Mutation analysis and protein interaction/expression studies in patient-derived material","journal":"Congenital Anomalies","confidence":"Low","confidence_rationale":"Tier 3 — single study in patient-derived samples, limited mechanistic detail available from abstract","pmids":["29845660"],"is_preprint":false},{"year":2023,"finding":"Homozygous frameshift deletions in EFCAB7 (loss-of-function) cause autosomal recessive nonsyndromic postaxial polydactyly in humans, consistent with EFCAB7's role as a positive regulator of Hedgehog signaling during limb formation via the IQCE-EFCAB7/EVC-EVC2 heterotetramer complex.","method":"Whole exome sequencing, Sanger sequencing segregation analysis in four consanguineous Pakistani families","journal":"European Journal of Human Genetics","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function variants with clear segregation in multiple families, mechanistic pathway placement from prior literature","pmids":["37684519"],"is_preprint":false},{"year":2025,"finding":"A splicing variant in EFCAB7 (c.683-1G>C) causes exon skipping and production of a non-functional protein. Mice carrying this variant show impaired ciliogenesis, disrupted Hedgehog/Shh-Gli pathway activity, and abnormal cardiac development (Tetralogy of Fallot-like defects). Transcriptional downregulation of Gli target genes including Myh6, Zfpm1, and Nkx2-5 was demonstrated as a downstream consequence of Shh signaling inhibition.","method":"Sanger sequencing of TOF patients, in vitro and in vivo splicing assays, mouse knockin model with cardiac phenotyping, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts","journal":"Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 2 — in vivo mouse model with multiple orthogonal methods (ciliogenesis assay, Hh pathway readouts, CUT&Tag, RNA-seq), clear mechanistic pathway from EFCAB7 loss to cardiac gene expression changes","pmids":["39894222"],"is_preprint":false},{"year":2024,"finding":"Complete deletion of Efcab7 in mice sensitizes embryos to Smoothened (Smo) antagonists (alcohol, vismodegib), causing more severe craniofacial and ocular malformations, while reducing malformations induced by a Smo agonist (SAG). This demonstrates that EFCAB7 facilitates Smoothened-dependent Shh pathway activity in vivo.","method":"CRISPR/Cas9 Efcab7 knockout mice, pharmacological challenge with Smo agonist/antagonist during gastrulation, fetal morphological phenotyping","journal":"Reproductive Toxicology","confidence":"Medium","confidence_rationale":"Tier 2 — genetic KO mouse with pharmacological epistasis clearly placing EFCAB7 in Smo-dependent Hh signaling","pmids":["39366525"],"is_preprint":false},{"year":2024,"finding":"EFCAB7 directly interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells, and this interaction promotes tumor cell proliferation and metastasis. Overexpression of PARK7 in EFCAB7-knockdown cells rescues proliferation and metastasis, indicating a functional EFCAB7-PARK7 axis in HCC.","method":"Mass spectrometry, Co-immunoprecipitation, siRNA knockdown, in vitro and in vivo proliferation/metastasis assays, PARK7 rescue overexpression","journal":"Aging","confidence":"Medium","confidence_rationale":"Tier 2/3 — reciprocal Co-IP and MS identification of PARK7, functional rescue experiment, but single lab in cancer cell context distinct from canonical ciliary function","pmids":["39276379"],"is_preprint":false},{"year":2025,"finding":"Efcab7 knockout mice generated by CRISPR/Cas9 are viable and fertile as males, with no significant differences in testicular morphology, weight, epididymis morphology, or sperm motility compared to wild-type, indicating Efcab7 is dispensable for male fertility when individually ablated.","method":"CRISPR/Cas9 knockout, mating tests, testis/epididymis histology, sperm motility analysis","journal":"Journal of Biomedical Research","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO with defined phenotypic readout (negative result), rigorously characterized","pmids":["41194443"],"is_preprint":false}],"current_model":"EFCAB7 functions as a core component of the EvC ciliary complex, where it forms a heterotetramer with IQCE, EVC, and EVC2 to anchor EVC-EVC2 in a signaling microdomain (the EvC zone) at the base of primary cilia, thereby facilitating Smoothened-dependent Hedgehog/Gli pathway activation; loss of EFCAB7 causes mislocalization of EVC-EVC2, impaired GLI2 activation, and downstream developmental defects including polydactyly and congenital heart disease, and EFCAB7 also interacts with PARK7 in a cancer context to promote hepatocellular carcinoma proliferation and metastasis."},"narrative":{"teleology":[{"year":2014,"claim":"The central question of how EVC-EVC2 is confined to the EvC zone at the ciliary base was answered by identifying EFCAB7-IQCE as the anchoring complex, establishing EFCAB7 as a required positive regulator of Hedgehog/Gli signaling.","evidence":"Reciprocal Co-IP, siRNA knockdown with ciliary localization imaging, GLI2 reporter assays, and domain-mapping of EVC2 binding in mammalian cells","pmids":["24582806"],"confidence":"High","gaps":["Structural basis of the EFCAB7-EVC2 interaction unresolved","Whether EFCAB7's EF-hand domains bind calcium and whether calcium regulates complex assembly unknown","In vivo developmental consequences of EFCAB7 loss not yet tested"]},{"year":2016,"claim":"Patient genetics provided the first human evidence that EFCAB7 variants modify EvC-related disease severity, linking EFCAB7 to Ellis-van Creveld syndrome as a genetic modifier.","evidence":"Sanger sequencing and SNP array analysis of an EVC patient carrying a heterozygous EFCAB7 missense variant (p.Y391H)","pmids":["26748586"],"confidence":"Low","gaps":["No functional reconstitution of the Y391H variant to demonstrate pathogenicity","Single patient observation without segregation in multiple families","Effect of the variant on EFCAB7-EVC2 binding not tested"]},{"year":2018,"claim":"A second EFCAB7 missense mutation was shown to alter expression levels and interactions among EvC complex components, reinforcing EFCAB7's role in complex stabilization.","evidence":"Mutation analysis and protein interaction/expression studies in patient-derived material","pmids":["29845660"],"confidence":"Low","gaps":["Limited mechanistic detail; no cell-biological or in vivo functional assays reported","Single study with patient-derived samples only"]},{"year":2023,"claim":"Two advances solidified EFCAB7's role: (1) homozygous loss-of-function frameshift deletions in EFCAB7 were shown to cause autosomal recessive postaxial polydactyly across multiple families, confirming EFCAB7 as an essential Hedgehog regulator for limb patterning; (2) post-translational modifications (SUMOylation enhancing and monoubiquitination reducing EVC-EVC2 EvC zone targeting) were mapped, and a second EFCAB7-binding motif in EVC2 was identified.","evidence":"Whole exome and Sanger sequencing with segregation in four consanguineous families; AP-MS interactomics, SUMO/ubiquitin modification assays, confocal microscopy, domain mapping","pmids":["37684519","37576597"],"confidence":"Medium","gaps":["Functional consequence of the second EFCAB7-binding motif versus the first not dissected independently","Whether SUMOylation directly modulates EFCAB7-EVC2 affinity or acts indirectly remains unclear","No rescue of polydactyly phenotype with EFCAB7 re-expression in patient cells"]},{"year":2024,"claim":"Mouse knockout studies established that EFCAB7 facilitates Smoothened-dependent Shh signaling in vivo: Efcab7-null embryos are hypersensitive to Smo antagonists and resistant to a Smo agonist, placing EFCAB7 genetically downstream of or at the level of Smoothened.","evidence":"CRISPR/Cas9 Efcab7 knockout mice challenged with vismodegib, alcohol (Smo antagonists) and SAG (Smo agonist) during gastrulation, with fetal morphological phenotyping","pmids":["39366525"],"confidence":"Medium","gaps":["Molecular mechanism by which EFCAB7 promotes Smo-dependent signaling (e.g., whether it affects Smo ciliary trafficking) not determined","Craniofacial phenotype penetrance and variability in unstressed Efcab7 KO mice not fully characterized"]},{"year":2024,"claim":"An unexpected non-ciliary role emerged: EFCAB7 directly interacts with PARK7/DJ-1 in hepatocellular carcinoma cells, and this axis promotes tumor proliferation and metastasis, revealing a context-dependent oncogenic function.","evidence":"Mass spectrometry identification, reciprocal Co-IP, siRNA knockdown with proliferation/metastasis assays, PARK7 rescue overexpression in HCC cell lines and xenografts","pmids":["39276379"],"confidence":"Medium","gaps":["Whether the EFCAB7-PARK7 interaction occurs through EFCAB7's EF-hand domains or a distinct interface is unknown","Relevance beyond HCC and relationship to ciliary Hedgehog function not explored","Single-lab finding not yet independently replicated"]},{"year":2025,"claim":"In vivo consequences of EFCAB7 loss on cardiac development were defined: a splicing variant causes impaired ciliogenesis, disrupted Shh-Gli activity, and Tetralogy of Fallot-like heart defects in a mouse knockin model, tracing the mechanism from EFCAB7 loss through reduced Gli binding at cardiac gene promoters to downregulation of Myh6, Zfpm1, and Nkx2-5.","evidence":"Mouse knockin model, splicing assays, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts, cardiac phenotyping","pmids":["39894222"],"confidence":"High","gaps":["Whether EFCAB7 loss affects ciliogenesis cell-autonomously or via secondary paracrine Hh signaling deficits not resolved","Human patient validation of the splicing variant limited to sequencing without functional studies in human cells"]},{"year":2025,"claim":"Male fertility is unaffected in Efcab7 knockout mice, demonstrating that EFCAB7 is dispensable for spermatogenesis and sperm motility despite primary cilia/flagella sharing structural components.","evidence":"CRISPR/Cas9 knockout mice with mating tests, testis/epididymis histology, sperm motility analysis","pmids":["41194443"],"confidence":"Medium","gaps":["Potential redundancy with other EF-hand proteins in sperm flagella not tested","Female fertility not assessed"]},{"year":null,"claim":"Key unresolved questions include the structural basis of EFCAB7-EVC2 interaction, whether EFCAB7's EF-hand domains bind calcium and how calcium might regulate EvC complex assembly, and the mechanism by which EFCAB7 engages PARK7 in non-ciliary contexts.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal or cryo-EM structure of the EFCAB7-containing complex","Calcium binding and its functional significance for EFCAB7 never demonstrated","Relationship between ciliary and oncogenic functions of EFCAB7 unexplored"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1,5]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,5,6]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,5,6]}],"complexes":["EvC ciliary complex (EFCAB7-IQCE-EVC-EVC2)"],"partners":["EVC2","EVC","IQCE","PARK7"],"other_free_text":[]},"mechanistic_narrative":"EFCAB7 is a core subunit of the EvC ciliary complex that facilitates Hedgehog/Gli pathway activation at primary cilia. It forms a heterotetramer with IQCE, EVC, and EVC2, directly binding a C-terminal disordered region of EVC2 to anchor the EVC-EVC2 module within the EvC zone at the ciliary base; depletion of EFCAB7 causes EVC-EVC2 mislocalization and impaired GLI2 activation [PMID:24582806, PMID:37576597]. Loss-of-function mutations in EFCAB7 cause autosomal recessive nonsyndromic postaxial polydactyly in humans [PMID:37684519], and a splicing variant produces impaired ciliogenesis, disrupted Shh-Gli signaling, and Tetralogy of Fallot-like cardiac defects in a mouse knockin model [PMID:39894222]. In a distinct context, EFCAB7 interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells to promote proliferation and metastasis [PMID:39276379]."},"prefetch_data":{"uniprot":{"accession":"A8K855","full_name":"EF-hand calcium-binding domain-containing protein 7","aliases":[],"length_aa":629,"mass_kda":72.0,"function":"Component of the EvC complex that positively regulates ciliary Hedgehog (Hh) signaling. Required for the localization of the EVC2:EVC subcomplex at the base of primary cilia","subcellular_location":"Cell projection, cilium membrane","url":"https://www.uniprot.org/uniprotkb/A8K855/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EFCAB7","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EFCAB7","total_profiled":1310},"omim":[{"mim_id":"617632","title":"EF-HAND CALCIUM-BINDING DOMAIN-CONTAINING PROTEIN 7; EFCAB7","url":"https://www.omim.org/entry/617632"},{"mim_id":"617631","title":"IQ DOMAIN-CONTAINING PROTEIN E; IQCE","url":"https://www.omim.org/entry/617631"},{"mim_id":"607261","title":"EVC CILIARY COMPLEX SUBUNIT 2; EVC2","url":"https://www.omim.org/entry/607261"},{"mim_id":"604831","title":"EVC CILIARY COMPLEX SUBUNIT 1; EVC","url":"https://www.omim.org/entry/604831"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Primary cilium","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EFCAB7"},"hgnc":{"alias_symbol":["KIAA1799","RP4-534K7.1"],"prev_symbol":[]},"alphafold":{"accession":"A8K855","domains":[{"cath_id":"1.10.238,1.10.238","chopping":"32-99","consensus_level":"medium","plddt":87.1854,"start":32,"end":99},{"cath_id":"1.10.238.10","chopping":"105-169","consensus_level":"medium","plddt":88.0805,"start":105,"end":169},{"cath_id":"2.60.40.1180","chopping":"245-258_527-629","consensus_level":"high","plddt":88.9703,"start":245,"end":629},{"cath_id":"2.60.120","chopping":"260-379_507-525","consensus_level":"medium","plddt":89.4173,"start":260,"end":525},{"cath_id":"1.10.238.10","chopping":"384-499","consensus_level":"medium","plddt":93.009,"start":384,"end":499}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/A8K855","model_url":"https://alphafold.ebi.ac.uk/files/AF-A8K855-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-A8K855-F1-predicted_aligned_error_v6.png","plddt_mean":80.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EFCAB7","jax_strain_url":"https://www.jax.org/strain/search?query=EFCAB7"},"sequence":{"accession":"A8K855","fasta_url":"https://rest.uniprot.org/uniprotkb/A8K855.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/A8K855/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/A8K855"}},"corpus_meta":[{"pmid":"24582806","id":"PMC_24582806","title":"EFCAB7 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EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of the transcription factor GLI2, mimicking the Weyers cellular phenotype.\",\n      \"method\": \"Co-immunoprecipitation, protein interaction mapping, shRNA depletion with ciliary localization readout, GLI2 activation assay\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, direct binding mapped to specific domain, loss-of-function with defined cellular and signaling phenotypes, replicated across multiple experiments\",\n      \"pmids\": [\"24582806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EFCAB7-IQCE complex interaction with EVC-EVC2 is enhanced by SUMOylation (SUMO3) of EVC-EVC2 cytosolic tails, promoting accumulation of EVC-EVC2 at the EvC zone. EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide, with a second motif mapped in this study.\",\n      \"method\": \"Endogenous EVC protein interactome proteomics in control and Evc-null cells, SUMO3 modification assays, ciliary localization imaging, domain mapping\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mass spectrometry interactome, confirmed known interactors (EVC2, IQCE, EFCAB7), functional SUMO modification assay with localization readout, domain mapping with mutagenesis\",\n      \"pmids\": [\"37576597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A splicing variant (c.683-1G>C) in EFCAB7 causes exon skipping and produces a non-functional protein. Mice carrying this variant show impaired ciliogenesis, disrupted Hedgehog/Shh-Gli pathway activity, and abnormal cardiac development. Downstream Gli target genes including Myh6, Zfpm1, and Nkx2-5 are transcriptionally downregulated as a consequence of Shh signaling inhibition caused by EFCAB7 loss.\",\n      \"method\": \"Sanger sequencing in patients, in vitro and in vivo splicing assays, mouse knock-in model with cardiac phenotype, CUT&Tag on Glis, bulk RNA-seq of embryonic hearts\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vivo mouse model with defined molecular phenotype, multiple orthogonal methods including CUT&Tag and RNA-seq confirming pathway position\",\n      \"pmids\": [\"39894222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EFCAB7 facilitates the actions of Smoothened (Smo) in the Sonic hedgehog pathway. Efcab7 knockout mice are more sensitive to Smo antagonist (vismodegib)-induced craniofacial malformations and less sensitive to Smo agonist (SAG)-induced malformations, placing EFCAB7 functionally at the level of Smo activation in the Shh pathway.\",\n      \"method\": \"Efcab7 knockout mouse model (Efcab7-/-), pharmacological challenge with Smo agonist/antagonist, craniofacial phenotype analysis\",\n      \"journal\": \"Reproductive toxicology (Elmsford, N.Y.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with genetic epistasis via pharmacological Smo modulation, clear phenotypic readout but single lab\",\n      \"pmids\": [\"39366525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EFCAB7 directly interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells. EFCAB7 promotes tumor cell proliferation and metastasis while inhibiting apoptosis, and overexpression of PARK7 rescues these functions in EFCAB7 knockdown cells, indicating PARK7 acts downstream of EFCAB7 in this context.\",\n      \"method\": \"Mass spectrometry, Co-immunoprecipitation, EFCAB7 knockdown with in vitro proliferation/metastasis/apoptosis assays, PARK7 rescue experiment\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — Co-IP and MS confirm direct interaction, rescue experiment places PARK7 downstream, but single lab and cancer-specific context distinct from canonical EFCAB7 ciliary function\",\n      \"pmids\": [\"39276379\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Efcab7 knockout (Efcab7-/-) male mice are fertile with no significant differences in testicular appearance, weight, testis/epididymis morphology, or sperm motility parameters compared to wild-type mice, demonstrating EFCAB7 is dispensable for male reproduction.\",\n      \"method\": \"CRISPR/Cas9 knockout in mice, mating tests, histological analysis of testis and epididymis, sperm motility measurement\",\n      \"journal\": \"Journal of biomedical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean CRISPR KO with multiple phenotypic readouts, single lab, negative result\",\n      \"pmids\": [\"41194443\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EFCAB7 forms a complex with IQCE that anchors the EVC-EVC2 heterodimer to the EvC zone at the base of primary cilia by binding to a C-terminal region of EVC2; this scaffolding function is required for proper Hedgehog/Smoothened-GLI signaling, and SUMO3 modification of EVC-EVC2 enhances their interaction with the EFCAB7-IQCE complex, while ubiquitination reduces EVC-EVC2 stability, collectively regulating Hh pathway output in skeletal, cardiac, and limb development.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2014,\n      \"finding\": \"EFCAB7 forms a complex with IQCE that anchors the EVC-EVC2 complex in a signaling microdomain at the base of primary cilia (the EvC zone). EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of the transcription factor GLI2, phenocopying the Weyers cellular phenotype.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, fluorescence microscopy/live imaging of ciliary localization, GLI2 reporter assays, evolutionary/bioinformatic analysis\",\n      \"journal\": \"Developmental Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, direct binding mapped to specific domain, KD with defined ciliary mislocalization phenotype and downstream signaling readout; replicated in part by subsequent studies\",\n      \"pmids\": [\"24582806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"SUMO3 modification of EVC-EVC2 cytosolic tails enhances their 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; only one was previously characterized, and the second was mapped in this study. Monoubiquitination of EVC-EVC2 cytosolic tails reduces their protein levels. Proteomic screen of the EVC interactome confirmed EVC2, IQCE, and EFCAB7 as core interactors.\",\n      \"method\": \"Endogenous protein interactome proteomics (AP-MS) in control and Evc-null cells, SUMO and ubiquitin modification assays, confocal microscopy of ciliary localization, domain mapping\",\n      \"journal\": \"Frontiers in Cell and Developmental Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — AP-MS interactome plus localization and modification assays from a single lab; moderate evidence for EFCAB7-binding motif mapping\",\n      \"pmids\": [\"37576597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"A heterozygous missense variant in EFCAB7 (c.1171T>C; p.Y391H) was identified in an Ellis-van Creveld patient also carrying an EVC truncation mutation, suggesting EFCAB7 acts as a modifier of EVC-related phenotypes through its interaction with EVC.\",\n      \"method\": \"Sanger sequencing, SNP array, mRNA expression analysis in patient samples\",\n      \"journal\": \"Congenital Anomalies\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — genetic/expression data from patient samples, no direct functional reconstitution\",\n      \"pmids\": [\"26748586\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A novel missense mutation in EFCAB7 was shown to alter the expression levels and interaction of Ellis-van Creveld ciliary proteins (EVC/EVC2/IQCE), supporting EFCAB7's role in stabilizing the EvC ciliary complex.\",\n      \"method\": \"Mutation analysis and protein interaction/expression studies in patient-derived material\",\n      \"journal\": \"Congenital Anomalies\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single study in patient-derived samples, limited mechanistic detail available from abstract\",\n      \"pmids\": [\"29845660\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Homozygous frameshift deletions in EFCAB7 (loss-of-function) cause autosomal recessive nonsyndromic postaxial polydactyly in humans, consistent with EFCAB7's role as a positive regulator of Hedgehog signaling during limb formation via the IQCE-EFCAB7/EVC-EVC2 heterotetramer complex.\",\n      \"method\": \"Whole exome sequencing, Sanger sequencing segregation analysis in four consanguineous Pakistani families\",\n      \"journal\": \"European Journal of Human Genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function variants with clear segregation in multiple families, mechanistic pathway placement from prior literature\",\n      \"pmids\": [\"37684519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A splicing variant in EFCAB7 (c.683-1G>C) causes exon skipping and production of a non-functional protein. Mice carrying this variant show impaired ciliogenesis, disrupted Hedgehog/Shh-Gli pathway activity, and abnormal cardiac development (Tetralogy of Fallot-like defects). Transcriptional downregulation of Gli target genes including Myh6, Zfpm1, and Nkx2-5 was demonstrated as a downstream consequence of Shh signaling inhibition.\",\n      \"method\": \"Sanger sequencing of TOF patients, in vitro and in vivo splicing assays, mouse knockin model with cardiac phenotyping, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo mouse model with multiple orthogonal methods (ciliogenesis assay, Hh pathway readouts, CUT&Tag, RNA-seq), clear mechanistic pathway from EFCAB7 loss to cardiac gene expression changes\",\n      \"pmids\": [\"39894222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Complete deletion of Efcab7 in mice sensitizes embryos to Smoothened (Smo) antagonists (alcohol, vismodegib), causing more severe craniofacial and ocular malformations, while reducing malformations induced by a Smo agonist (SAG). This demonstrates that EFCAB7 facilitates Smoothened-dependent Shh pathway activity in vivo.\",\n      \"method\": \"CRISPR/Cas9 Efcab7 knockout mice, pharmacological challenge with Smo agonist/antagonist during gastrulation, fetal morphological phenotyping\",\n      \"journal\": \"Reproductive Toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO mouse with pharmacological epistasis clearly placing EFCAB7 in Smo-dependent Hh signaling\",\n      \"pmids\": [\"39366525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EFCAB7 directly interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells, and this interaction promotes tumor cell proliferation and metastasis. Overexpression of PARK7 in EFCAB7-knockdown cells rescues proliferation and metastasis, indicating a functional EFCAB7-PARK7 axis in HCC.\",\n      \"method\": \"Mass spectrometry, Co-immunoprecipitation, siRNA knockdown, in vitro and in vivo proliferation/metastasis assays, PARK7 rescue overexpression\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — reciprocal Co-IP and MS identification of PARK7, functional rescue experiment, but single lab in cancer cell context distinct from canonical ciliary function\",\n      \"pmids\": [\"39276379\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Efcab7 knockout mice generated by CRISPR/Cas9 are viable and fertile as males, with no significant differences in testicular morphology, weight, epididymis morphology, or sperm motility compared to wild-type, indicating Efcab7 is dispensable for male fertility when individually ablated.\",\n      \"method\": \"CRISPR/Cas9 knockout, mating tests, testis/epididymis histology, sperm motility analysis\",\n      \"journal\": \"Journal of Biomedical Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined phenotypic readout (negative result), rigorously characterized\",\n      \"pmids\": [\"41194443\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EFCAB7 functions as a core component of the EvC ciliary complex, where it forms a heterotetramer with IQCE, EVC, and EVC2 to anchor EVC-EVC2 in a signaling microdomain (the EvC zone) at the base of primary cilia, thereby facilitating Smoothened-dependent Hedgehog/Gli pathway activation; loss of EFCAB7 causes mislocalization of EVC-EVC2, impaired GLI2 activation, and downstream developmental defects including polydactyly and congenital heart disease, and EFCAB7 also interacts with PARK7 in a cancer context to promote hepatocellular carcinoma proliferation and metastasis.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"EFCAB7 is a ciliary scaffolding protein that, together with IQCE, anchors the EVC-EVC2 signaling complex to the EvC zone at the base of primary cilia, thereby enabling Hedgehog/Smoothened-to-GLI signal transduction required for skeletal, craniofacial, and cardiac development. EFCAB7 directly binds a C-terminal disordered region of EVC2 through two distinct motifs within the peptide deleted in Weyers syndrome patients, and SUMOylation of EVC-EVC2 enhances this interaction to promote EvC zone accumulation [PMID:24582806, PMID:37576597]. Loss of EFCAB7 mislocalizes EVC-EVC2 within cilia and impairs GLI2 activation, and Efcab7 knockout mice show heightened sensitivity to Smoothened antagonists and reduced sensitivity to Smoothened agonists, placing EFCAB7 functionally at the level of Smo activation [PMID:24582806, PMID:39366525]. A loss-of-function splicing variant in EFCAB7 causes impaired ciliogenesis and disrupted Shh-Gli signaling leading to abnormal cardiac development in mice, with transcriptional downregulation of Gli target genes including Myh6, Nkx2-5, and Zfpm1 [PMID:39894222].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"The central mechanistic question — how EVC-EVC2 is localized to the ciliary base signaling domain — was answered by identifying the EFCAB7-IQCE complex as its anchor, establishing EFCAB7 as a critical scaffold in Hedgehog signal transduction.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, domain interaction mapping to EVC2 C-terminus, shRNA depletion with ciliary localization and GLI2 activation readouts in mammalian cells\",\n      \"pmids\": [\"24582806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of the EFCAB7-EVC2 interaction not resolved\",\n        \"Whether EFCAB7 has direct enzymatic or signaling activity beyond scaffolding unknown\",\n        \"In vivo phenotypic consequences of EFCAB7 loss not yet tested in animal models\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"The regulatory mechanism governing EVC-EVC2 accumulation at the EvC zone was clarified: SUMO3 modification of EVC-EVC2 cytosolic tails enhances their interaction with the EFCAB7-IQCE complex, and a second EFCAB7-binding motif within the Weyers-deleted peptide of EVC2 was mapped.\",\n      \"evidence\": \"Endogenous EVC interactome proteomics in control and Evc-null cells, SUMO3 modification assays with ciliary localization imaging, mutagenesis-based domain mapping\",\n      \"pmids\": [\"37576597\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Which SUMO E3 ligase modifies EVC-EVC2 in this context remains unidentified\",\n        \"Relative contribution of each EFCAB7-binding motif to EvC zone targeting not quantified\",\n        \"Whether deSUMOylation dynamically regulates the complex during active signaling is untested\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Genetic epistasis in vivo placed EFCAB7 functionally at the level of Smoothened activation: Efcab7 knockout mice were hypersensitive to Smo antagonism and hyposensitive to Smo agonism, resolving where in the pathway EFCAB7 acts.\",\n      \"evidence\": \"Efcab7 knockout mouse model challenged with vismodegib (Smo antagonist) and SAG (Smo agonist), craniofacial phenotype scoring\",\n      \"pmids\": [\"39366525\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single laboratory finding; independent replication in a second lab not yet reported\",\n        \"Biochemical mechanism by which EFCAB7 facilitates Smo signaling (beyond EVC-EVC2 anchoring) remains undefined\",\n        \"Whether craniofacial defects reflect cell-autonomous ciliary dysfunction versus broader developmental perturbation not distinguished\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"A potential non-ciliary function was uncovered: EFCAB7 interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells, and PARK7 overexpression rescues proliferation and metastatic phenotypes caused by EFCAB7 knockdown, suggesting a PARK7-dependent oncogenic role.\",\n      \"evidence\": \"Mass spectrometry and co-immunoprecipitation in HCC cell lines, EFCAB7 knockdown with PARK7 rescue in proliferation, metastasis, and apoptosis assays\",\n      \"pmids\": [\"39276379\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Cancer-specific context distinct from canonical ciliary function; whether this reflects a genuine non-ciliary mechanism or an artifact of transformed cell biology is unclear\",\n        \"Direct binding domain on EFCAB7 for PARK7 not mapped\",\n        \"No in vivo tumor model validation\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"In vivo cardiac phenotyping resolved that EFCAB7 is essential for heart development: a loss-of-function splicing variant impairs ciliogenesis and Shh-Gli signaling, downregulating cardiac transcription factor genes (Myh6, Nkx2-5, Zfpm1), while male reproductive function is dispensable.\",\n      \"evidence\": \"Mouse knock-in of patient splicing variant (c.683-1G>C), CUT&Tag for Gli occupancy, RNA-seq of embryonic hearts; separate CRISPR KO with testis histology and sperm motility analysis\",\n      \"pmids\": [\"39894222\", \"41194443\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether EFCAB7 loss-of-function causes human congenital heart disease remains to be confirmed in patient cohorts\",\n        \"Cell-type specificity of ciliogenesis defect (cardiomyocyte progenitors vs. other lineages) not resolved\",\n        \"Phenotypic consequences in female reproductive function not assessed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis of EFCAB7-EVC2 interaction, whether EFCAB7 has EF-hand calcium-dependent regulatory activity, the physiological relevance of the EFCAB7-PARK7 interaction outside cancer, and whether EFCAB7 mutations cause Mendelian disease in humans.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model (crystal or cryo-EM) of the EFCAB7-IQCE-EVC2 complex exists\",\n        \"Calcium-binding capacity of EFCAB7 EF-hand domains has never been tested\",\n        \"Human genetic studies linking EFCAB7 mutations to congenital disease are lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 3]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [\n      \"EFCAB7-IQCE complex\",\n      \"EVC-EVC2-EFCAB7-IQCE complex\"\n    ],\n    \"partners\": [\n      \"IQCE\",\n      \"EVC2\",\n      \"EVC\",\n      \"PARK7\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"EFCAB7 is a core subunit of the EvC ciliary complex that facilitates Hedgehog/Gli pathway activation at primary cilia. It forms a heterotetramer with IQCE, EVC, and EVC2, directly binding a C-terminal disordered region of EVC2 to anchor the EVC-EVC2 module within the EvC zone at the ciliary base; depletion of EFCAB7 causes EVC-EVC2 mislocalization and impaired GLI2 activation [PMID:24582806, PMID:37576597]. Loss-of-function mutations in EFCAB7 cause autosomal recessive nonsyndromic postaxial polydactyly in humans [PMID:37684519], and a splicing variant produces impaired ciliogenesis, disrupted Shh-Gli signaling, and Tetralogy of Fallot-like cardiac defects in a mouse knockin model [PMID:39894222]. In a distinct context, EFCAB7 interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells to promote proliferation and metastasis [PMID:39276379].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"The central question of how EVC-EVC2 is confined to the EvC zone at the ciliary base was answered by identifying EFCAB7-IQCE as the anchoring complex, establishing EFCAB7 as a required positive regulator of Hedgehog/Gli signaling.\",\n      \"evidence\": \"Reciprocal Co-IP, siRNA knockdown with ciliary localization imaging, GLI2 reporter assays, and domain-mapping of EVC2 binding in mammalian cells\",\n      \"pmids\": [\"24582806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of the EFCAB7-EVC2 interaction unresolved\",\n        \"Whether EFCAB7's EF-hand domains bind calcium and whether calcium regulates complex assembly unknown\",\n        \"In vivo developmental consequences of EFCAB7 loss not yet tested\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Patient genetics provided the first human evidence that EFCAB7 variants modify EvC-related disease severity, linking EFCAB7 to Ellis-van Creveld syndrome as a genetic modifier.\",\n      \"evidence\": \"Sanger sequencing and SNP array analysis of an EVC patient carrying a heterozygous EFCAB7 missense variant (p.Y391H)\",\n      \"pmids\": [\"26748586\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No functional reconstitution of the Y391H variant to demonstrate pathogenicity\",\n        \"Single patient observation without segregation in multiple families\",\n        \"Effect of the variant on EFCAB7-EVC2 binding not tested\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"A second EFCAB7 missense mutation was shown to alter expression levels and interactions among EvC complex components, reinforcing EFCAB7's role in complex stabilization.\",\n      \"evidence\": \"Mutation analysis and protein interaction/expression studies in patient-derived material\",\n      \"pmids\": [\"29845660\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Limited mechanistic detail; no cell-biological or in vivo functional assays reported\",\n        \"Single study with patient-derived samples only\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Two advances solidified EFCAB7's role: (1) homozygous loss-of-function frameshift deletions in EFCAB7 were shown to cause autosomal recessive postaxial polydactyly across multiple families, confirming EFCAB7 as an essential Hedgehog regulator for limb patterning; (2) post-translational modifications (SUMOylation enhancing and monoubiquitination reducing EVC-EVC2 EvC zone targeting) were mapped, and a second EFCAB7-binding motif in EVC2 was identified.\",\n      \"evidence\": \"Whole exome and Sanger sequencing with segregation in four consanguineous families; AP-MS interactomics, SUMO/ubiquitin modification assays, confocal microscopy, domain mapping\",\n      \"pmids\": [\"37684519\", \"37576597\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional consequence of the second EFCAB7-binding motif versus the first not dissected independently\",\n        \"Whether SUMOylation directly modulates EFCAB7-EVC2 affinity or acts indirectly remains unclear\",\n        \"No rescue of polydactyly phenotype with EFCAB7 re-expression in patient cells\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mouse knockout studies established that EFCAB7 facilitates Smoothened-dependent Shh signaling in vivo: Efcab7-null embryos are hypersensitive to Smo antagonists and resistant to a Smo agonist, placing EFCAB7 genetically downstream of or at the level of Smoothened.\",\n      \"evidence\": \"CRISPR/Cas9 Efcab7 knockout mice challenged with vismodegib, alcohol (Smo antagonists) and SAG (Smo agonist) during gastrulation, with fetal morphological phenotyping\",\n      \"pmids\": [\"39366525\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which EFCAB7 promotes Smo-dependent signaling (e.g., whether it affects Smo ciliary trafficking) not determined\",\n        \"Craniofacial phenotype penetrance and variability in unstressed Efcab7 KO mice not fully characterized\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"An unexpected non-ciliary role emerged: EFCAB7 directly interacts with PARK7/DJ-1 in hepatocellular carcinoma cells, and this axis promotes tumor proliferation and metastasis, revealing a context-dependent oncogenic function.\",\n      \"evidence\": \"Mass spectrometry identification, reciprocal Co-IP, siRNA knockdown with proliferation/metastasis assays, PARK7 rescue overexpression in HCC cell lines and xenografts\",\n      \"pmids\": [\"39276379\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether the EFCAB7-PARK7 interaction occurs through EFCAB7's EF-hand domains or a distinct interface is unknown\",\n        \"Relevance beyond HCC and relationship to ciliary Hedgehog function not explored\",\n        \"Single-lab finding not yet independently replicated\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"In vivo consequences of EFCAB7 loss on cardiac development were defined: a splicing variant causes impaired ciliogenesis, disrupted Shh-Gli activity, and Tetralogy of Fallot-like heart defects in a mouse knockin model, tracing the mechanism from EFCAB7 loss through reduced Gli binding at cardiac gene promoters to downregulation of Myh6, Zfpm1, and Nkx2-5.\",\n      \"evidence\": \"Mouse knockin model, splicing assays, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts, cardiac phenotyping\",\n      \"pmids\": [\"39894222\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether EFCAB7 loss affects ciliogenesis cell-autonomously or via secondary paracrine Hh signaling deficits not resolved\",\n        \"Human patient validation of the splicing variant limited to sequencing without functional studies in human cells\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Male fertility is unaffected in Efcab7 knockout mice, demonstrating that EFCAB7 is dispensable for spermatogenesis and sperm motility despite primary cilia/flagella sharing structural components.\",\n      \"evidence\": \"CRISPR/Cas9 knockout mice with mating tests, testis/epididymis histology, sperm motility analysis\",\n      \"pmids\": [\"41194443\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Potential redundancy with other EF-hand proteins in sperm flagella not tested\",\n        \"Female fertility not assessed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis of EFCAB7-EVC2 interaction, whether EFCAB7's EF-hand domains bind calcium and how calcium might regulate EvC complex assembly, and the mechanism by which EFCAB7 engages PARK7 in non-ciliary contexts.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No crystal or cryo-EM structure of the EFCAB7-containing complex\",\n        \"Calcium binding and its functional significance for EFCAB7 never demonstrated\",\n        \"Relationship between ciliary and oncogenic functions of EFCAB7 unexplored\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 5, 6]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 5, 6]}\n    ],\n    \"complexes\": [\n      \"EvC ciliary complex (EFCAB7-IQCE-EVC-EVC2)\"\n    ],\n    \"partners\": [\n      \"EVC2\",\n      \"EVC\",\n      \"IQCE\",\n      \"PARK7\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}