{"gene":"CCDC88C","run_date":"2026-06-09T22:57:17","timeline":{"discoveries":[{"year":2015,"finding":"DAPLE contains a Gα-binding and activating (GBA) motif that directly activates Gαi proteins, and an adjacent domain that directly binds Frizzled receptors (FZDRs), thereby linking Wnt stimulation to G protein activation. This triggers non-canonical Wnt responses: suppression of the β-catenin/TCF/LEF pathway and tumorigenesis, but enhancement of PI3K-Akt and Rac1 signals and tumor cell invasiveness.","method":"Biochemical binding assays, GEF activity assays, cell-based signaling assays, mutagenesis, tumor invasion models","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods including in vitro GEF assays, direct binding studies, mutagenesis of the GBA motif, and functional cellular readouts; foundational paper replicated by subsequent studies","pmids":["26126266"],"is_preprint":false},{"year":2012,"finding":"DAPLE interacts directly with Dishevelled (Dvl) and regulates Wnt5a-mediated activation of Rac and formation of lamellipodia. DAPLE increases the association of Dvl with an isoform of atypical protein kinase C (aPKC/PKCλ), promoting Rac activation, and is required for fibroblast and epithelial cell migration during wound healing in vivo.","method":"Co-immunoprecipitation, lamellipodia formation assays, Rac activation assays, siRNA knockdown, in vivo wound healing model in mice","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in vivo knockout phenotype, Rac activation assays, multiple cell types tested; replicated in subsequent studies","pmids":["22643886"],"is_preprint":false},{"year":2017,"finding":"DAPLE-deficient mice develop hydrocephalus with ependymal cilia lacking coordinated orientation. DAPLE regulates microtubule dynamics at the anterior side of ependymal cells, orienting cilial basal bodies required for directional cerebrospinal fluid (CSF) flow, establishing a role for DAPLE in planar polarity of motile cilia.","method":"Daple-deficient (knockout) mouse model, cilia orientation analysis, microtubule dynamics imaging, CSF flow assay","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with defined cellular phenotype (cilia disorientation, hydrocephalus), direct imaging of microtubule dynamics; replicated across multiple labs in different contexts","pmids":["28746879"],"is_preprint":false},{"year":2014,"finding":"A missense mutation in CCDC88C (p.R464H) activates the c-Jun N-terminal kinase (JNK) pathway, induces caspase-3 cleavage, and triggers apoptosis, linking DAPLE to JNK stress signaling and cerebellar neurodegeneration.","method":"Whole-exome sequencing, expression of mutant CCDC88C in cells, JNK pathway activation assay, caspase-3 cleavage assay, apoptosis assay","journal":"Journal of medical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-based functional assays with mutant protein and multiple readouts (JNK, caspase-3, apoptosis) in a single lab","pmids":["25062847"],"is_preprint":false},{"year":2018,"finding":"DAPLE is a substrate of multiple receptor tyrosine kinases (RTKs) and non-RTKs. Phosphorylation near the Dvl-binding motif causes DAPLE/Dvl complex dissociation and augments DAPLE's ability to bind to and activate Gαi, potentiating β-catenin-independent Wnt signals and stimulating epithelial-mesenchymal transition (EMT).","method":"Phosphoproteomics (mass spectrometry), Co-immunoprecipitation, in vitro kinase assay, Gαi activation assay, EMT assay","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — mass spectrometry identification of phosphorylation sites, in vitro kinase assays, Co-IP, Gαi nucleotide exchange assay, multiple orthogonal methods","pmids":["29487190"],"is_preprint":false},{"year":2019,"finding":"DAPLE directly binds the PDZ3 domain of MPDZ via its PDZ-binding motif (PBM) and colocalizes with MPDZ at apical cell junctions. MPDZ depletion blunts DAPLE-mediated apical constriction of cultured cells. Both proteins are required for apical constriction of neuroepithelial cells and neural plate bending during Xenopus neurulation.","method":"Co-immunoprecipitation, direct binding assay (PDZ domain pulldown), colocalization imaging, Xenopus morpholino knockdown, cell constriction assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct binding demonstrated between defined domains, functional rescue in Xenopus, cellular apical constriction assay, multiple orthogonal methods","pmids":["31268831"],"is_preprint":false},{"year":2017,"finding":"DAPLE interacts with both PCP (planar cell polarity) and cell-intrinsic G-protein-based signals in inner-ear hair cells. DAPLE is required to maintain the polarized distribution of the core PCP protein Dishevelled and to position the primary cilium at the abneural edge of the apical surface, integrating organ-wide and cell-intrinsic polarity.","method":"Mouse knockout/mutant analysis, immunofluorescence localization of PCP proteins, cilium positioning assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with defined cellular phenotype (Dvl mislocalization, cilium mispositioning), single lab","pmids":["29229865"],"is_preprint":false},{"year":2017,"finding":"Akt phosphorylates the phosphoinositide (PI)-binding domain of DAPLE, abolishing DAPLE's ability to bind PI3-P-enriched endosomes and engage dynein motor complex for trafficking of β-catenin/E-cadherin complexes to pericentriolar recycling endosomes (PCREs). Phosphorylation compartmentalizes DAPLE/β-catenin/E-cadherin complexes to cell-cell contact sites, enhancing non-canonical Wnt signals and suppressing colony growth.","method":"Phosphorylation assay, endosomal fractionation, Co-immunoprecipitation, colony growth assay, mutagenesis of PI-binding domain and Akt phosphorylation site","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct phosphorylation assay, fractionation, mutant analysis, single lab with multiple orthogonal methods","pmids":["29021338"],"is_preprint":false},{"year":2020,"finding":"DAPLE's GBA motif binds efficiently to Gαs and Gαq (in addition to Gαi) and inhibits nucleotide exchange on Gαs and Gαq, in contrast to the nucleotide-exchange acceleration it causes for Gαi. Met-1669 in the GBA motif was identified as the residue that enables better binding to Gαs and Gαq.","method":"In vitro nucleotide exchange assay, binding assay, site-directed mutagenesis (M1669 substitution)","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical reconstitution with mutagenesis defining mechanism; single lab but with rigorous in vitro assays","pmids":["31949046"],"is_preprint":false},{"year":2020,"finding":"TRIM11 associates with DAPLE and promotes its ubiquitin-mediated degradation via a p62-selective autophagic pathway, thereby upregulating β-catenin and inducing ABCC9 expression in nasopharyngeal carcinoma drug-resistant cells.","method":"Co-immunoprecipitation, ubiquitination assay, selective autophagy assay, β-catenin reporter assay","journal":"Oncogenesis","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP showing TRIM11–DAPLE interaction, ubiquitination assay, autophagic degradation shown, single lab with multiple assays","pmids":["32382014"],"is_preprint":false},{"year":2020,"finding":"DAPLE forms a trimeric complex with Dvl2 and CK1ε. Daple overexpression induces CK1ε-mediated phosphorylation of Dvl2 at Thr224, which is required for full activation of β-catenin transcriptional activity. A Daple mutant lacking the C-terminal GCV motif (Dvl-binding) retains CK1ε interaction but fails to induce Dvl phosphorylation. Wnt3a stimulation increases membrane localization of DAPLE and its association with Dvl.","method":"Co-immunoprecipitation, site-directed mutagenesis (ΔGCV, Dvl2-T224A), β-catenin reporter assay, membrane fractionation","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, mutagenesis of interaction domains, functional reporter assay, single lab","pmids":["32888647"],"is_preprint":false},{"year":2020,"finding":"DAPLE localizes to epithelial cell junctions via its PDZ-binding motif (PBM) binding the third PDZ domain of PARD3. Tyrosine phosphorylation of DAPLE's PBM by receptor and non-receptor tyrosine kinases (including Src) regulates the interaction: hypophosphorylation strengthens DAPLE–PARD3 interaction, while hyperphosphorylation disrupts it, controlling contact-triggered planar polarity.","method":"Co-immunoprecipitation, direct binding assay (PDZ domain pulldown), tyrosine kinase assay (Src), immunofluorescence localization, phosphomimetic/phosphonull mutagenesis","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct PDZ binding assay, phosphorylation by Src demonstrated, PBM mutagenesis, multiple methods in single lab","pmids":["32058970"],"is_preprint":false},{"year":2022,"finding":"The PAR polarity complex (PAR3-PAR6-aPKC) at apical cell junctions recruits DAPLE, which in turn triggers a two-pronged mechanism for assembly of the apical actomyosin network: (1) direct recruitment of the actin-stabilizing protein CD2AP to apical junctions, and (2) GPCR-independent activation of heterotrimeric G protein signaling to favor RhoA-myosin activation.","method":"Co-immunoprecipitation, immunofluorescence, DAPLE knockdown/overexpression, Rho activation assay, actomyosin assembly assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, functional knockdown with defined actomyosin phenotype, G protein activation assay, multiple orthogonal methods in one study","pmids":["35389423"],"is_preprint":false},{"year":2024,"finding":"DAPLE (CCDC88C) is a substrate of GALNT6, which promotes O-linked GalNAc glycosylation of DAPLE, maintaining its protein stability. This O-GalNAc modification is required for DAPLE's pro-metastatic function. DAPLE promotes breast cancer metastasis via c-JUN-mediated transcriptional activation of CEMIP.","method":"Co-immunoprecipitation, O-glycosylation assay, GALNT6 knockdown/overexpression, CCDC88C gain/loss-of-function, c-JUN reporter assay, in vivo metastasis model","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — O-glycosylation biochemically demonstrated, functional stability assay, in vivo metastasis model, single lab","pmids":["38971758"],"is_preprint":false},{"year":2015,"finding":"DAPLE depletion suppresses Wnt5a-induced Rac and JNK activation, laminin γ2 expression, and cell migration/invasion in gastric cancer cells. Daple depletion also suppressed liver metastasis in a mouse xenograft model, positioning DAPLE downstream of Wnt5a/Dvl to activate Rac and JNK in gastric cancer.","method":"siRNA knockdown, Rac activation assay, JNK phosphorylation assay, laminin γ2 expression assay, Transwell migration/invasion assay, mouse xenograft metastasis model","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with multiple defined pathway readouts and in vivo validation, single lab","pmids":["26577606"],"is_preprint":false},{"year":2005,"finding":"CCDC88C (KIAA1509) contains a coiled-coil oligomerization domain with homology to the HOOK family of proteins. The KIAA1509-PDGFRβ fusion protein resulting from t(5;14)(q33;q32) retains the KIAA1509 coiled-coil domain fused to the cytoplasmic kinase domain of PDGFRβ, producing a constitutively active tyrosine kinase that transforms hematopoietic cells.","method":"Inverse PCR cloning, sequencing, imatinib treatment (clinical response), domain analysis","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — fusion gene cloned and characterized; imatinib clinical response confirms constitutive kinase activity; replicated in multiple subsequent cases","pmids":["15496975"],"is_preprint":false},{"year":2025,"finding":"The CCDC88C-FLT3 (Daple-FLT3) fusion oncoprotein contains a constitutively active FLT3 kinase domain that activates STAT5a, AKT, and MAPK signaling without ligand stimulation. The coiled-coil domain of DAPLE is dispensable for kinase activation but necessary for maximal activation and pericentrosomal localization of the fusion protein. Sorafenib (and to a lesser degree imatinib) can modulate this signaling.","method":"Cell-based kinase activity assay, STAT5/AKT/MAPK phosphorylation assay, subcellular localization imaging, coiled-coil domain deletion mutagenesis, TKI treatment","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-based functional assay with domain mutagenesis and localization imaging; preprint, single lab","pmids":["41256517"],"is_preprint":true},{"year":2025,"finding":"DAPLE and its paralog Girdin-L bear unique extended C-terminal PDZ-binding motifs that bind to the PDZ domain of DVL2 with exceptionally high affinity, as revealed by proximity labeling combined with structural and biophysical analysis. Deletion of these motifs or the DVL2 PDZ domain results in elongated primary cilia and renders cilia unresponsive to Wnt5a-stimulated disassembly, establishing that the DVL2 PDZ–DAPLE interaction mediates primary cilia disassembly in response to Wnt5a.","method":"Proximity labeling (BioID), structural analysis, biophysical binding assay, PDZ motif deletion mutagenesis, primary cilia length measurement, Wnt5a stimulation assay in HEK293T cells","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — structural and biophysical binding with mutagenesis, functional cilia assay; preprint, single lab, not yet peer-reviewed","pmids":["bio_10.1101_2025.09.26.678488"],"is_preprint":true},{"year":2019,"finding":"Two isoforms of DAPLE/CCDC88C both suppress tumor growth via their GBA motif, but only the full-length isoform triggers EMT and invasion. Both isoforms are suppressed during colon cancer progression, and their reduced expression carries additive prognostic significance.","method":"Isoform cloning, GBA motif mutagenesis, colony formation assay, invasion assay","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional mutagenesis of GBA motif, isoform-specific gain-of-function, multiple cellular readouts, single lab","pmids":["31431650"],"is_preprint":false},{"year":2021,"finding":"Adult Daple-knockout mice exhibit hearing disturbances across a broad frequency range. In these mice, apical microtubules in cochlear hair cells are irregularly aggregated and the number of microtubules attached to plasma membranes is decreased, causing distorted hair bundle arrangement. Nocodazole treatment in wild-type cochlea culture phenocopied these microtubule defects, establishing DAPLE's role in apical microtubule organization and hair bundle patterning.","method":"Daple knockout mouse audiometry (auditory brainstem response), immunofluorescence of hair bundles, electron/confocal microscopy of apical microtubules, nocodazole pharmacological experiment","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout with defined audiological and cellular phenotype, pharmacological validation, single lab","pmids":["34642354"],"is_preprint":false},{"year":2010,"finding":"A homozygous splice-site mutation in CCDC88C (intron 29) causes loss of DAPLE protein. In blood from the affected patient, CTNNB1 (β-catenin) and LEF1 mRNA levels were increased compared to controls, consistent with loss of DAPLE's inhibitory role on canonical Wnt/β-catenin signaling.","method":"Western blotting (protein loss), quantitative RT-PCR of Wnt pathway genes, positional cloning","journal":"Molecular syndromology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single family, mRNA expression changes only (no direct pathway manipulation), no rescue experiment","pmids":["21031079"],"is_preprint":false},{"year":2012,"finding":"Mutations truncating the extreme C-terminus of DAPLE, including the GCV PDZ-binding motif, cause autosomal recessive congenital hydrocephalus, indicating that the PDZ domain-binding motif (which mediates Dishevelled binding) is functionally essential for DAPLE's role in brain development.","method":"Homozygosity mapping, whole exome sequencing, protein domain analysis of truncating mutations","journal":"Journal of medical genetics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — genetic evidence from human families; no direct in vitro functional assay; mechanistic inference from domain knowledge","pmids":["23042809"],"is_preprint":false}],"current_model":"DAPLE/CCDC88C is a multi-domain cytoplasmic scaffold and non-receptor guanine nucleotide exchange factor (GEF) that bridges Wnt/Frizzled receptor signaling to heterotrimeric G protein activation: its GBA motif activates Gαi while inhibiting Gαs and Gαq, its C-terminal PDZ-binding motif (GCV) docks to the PDZ domain of Dishevelled (DVL2) and PARD3/MPDZ at apical junctions, and it is regulated by phosphorylation from RTKs/Src (which dissociates the Dvl complex and augments Gαi activation) and by Akt (which controls endosomal trafficking of β-catenin); through these mechanisms DAPLE coordinates non-canonical Wnt-driven Rac activation and cell migration, apical actomyosin assembly via CD2AP recruitment and RhoA activation, planar polarity in ependymal and hair cells via apical microtubule organization, primary cilia disassembly in response to Wnt5a, and CK1ε-mediated Dvl2 phosphorylation that activates canonical β-catenin signaling, while its dysregulation—through loss-of-function mutations, gain-of-function JNK-activating mutations, oncogenic CCDC88C-PDGFRB/FLT3/JAK2 chromosomal fusions, or TRIM11-mediated autophagic degradation—underlies congenital hydrocephalus, spinocerebellar ataxia, and cancer progression."},"narrative":{"mechanistic_narrative":"CCDC88C/DAPLE is a multi-domain cytoplasmic scaffold that couples Wnt/Frizzled signaling to heterotrimeric G protein activation through a Gα-binding and activating (GBA) motif that accelerates nucleotide exchange on Gαi while binding and inhibiting nucleotide exchange on Gαs and Gαq, the latter selectivity conferred by Met-1669 [PMID:26126266, PMID:31949046]. By engaging Frizzled and Dishevelled, DAPLE drives non-canonical Wnt responses—suppressing β-catenin/TCF/LEF output while potentiating PI3K-Akt and Rac1 signaling, lamellipodia formation, and cell migration—and acts downstream of Wnt5a/Dvl to activate Rac and JNK [PMID:26126266, PMID:22643886, PMID:26577606]. Its C-terminal PDZ-binding motif docks DAPLE at apical junctions through high-affinity interactions with the PDZ domains of DVL2, PARD3, and MPDZ, where it organizes epithelial polarity and apical actomyosin assembly: recruited by the PAR3-PAR6-aPKC complex, DAPLE both recruits the actin-stabilizing protein CD2AP and triggers GPCR-independent RhoA-myosin activation [PMID:31268831, PMID:32058970, PMID:35389423]. DAPLE governs planar polarity and ciliary architecture, orienting ependymal cilia and positioning the primary cilium via apical microtubule organization [PMID:28746879, PMID:29229865, PMID:34642354]. DAPLE activity is gated by post-translational regulation: tyrosine kinases including Src phosphorylate sites near the Dvl- and PARD3-binding motifs to dissociate these complexes and augment Gαi activation and EMT, Akt phosphorylates its phosphoinositide-binding domain to control endosomal trafficking of β-catenin/E-cadherin complexes, and protein levels are set by GALNT6-mediated O-GalNAc glycosylation and by TRIM11-driven selective autophagic degradation [PMID:29487190, PMID:29021338, PMID:32382014, PMID:38971758]. Human loss-of-function and C-terminal truncating mutations in CCDC88C cause autosomal recessive congenital hydrocephalus, and a p.R464H missense mutation that activates JNK and apoptosis is linked to cerebellar neurodegeneration [PMID:23042809, PMID:25062847]. Distinct from the full-length signaling protein, the CCDC88C coiled-coil oligomerization domain forms constitutively active oncogenic kinase fusions with PDGFRβ and FLT3 that transform hematopoietic cells [PMID:15496975, PMID:41256517].","teleology":[{"year":2005,"claim":"Before its signaling role was known, CCDC88C was identified as a fusion partner whose coiled-coil domain could constitutively activate a kinase, establishing the first disease link via an oncogenic translocation.","evidence":"Inverse PCR cloning and sequencing of a t(5;14) KIAA1509-PDGFRβ fusion with imatinib clinical response","pmids":["15496975"],"confidence":"Medium","gaps":["Did not address the normal cellular function of full-length CCDC88C","Mechanism of oligomerization-driven kinase activation not structurally resolved"]},{"year":2010,"claim":"A human splice-site mutation causing DAPLE protein loss first implicated the gene in restraining canonical Wnt output, raising elevated β-catenin and LEF1 transcripts.","evidence":"Positional cloning, Western blot of protein loss, and qRT-PCR of Wnt pathway genes in a single family","pmids":["21031079"],"confidence":"Low","gaps":["Single family with mRNA correlation only, no pathway manipulation or rescue","Direct biochemical link to β-catenin regulation not established"]},{"year":2012,"claim":"Linking DAPLE to Dishevelled established it as a non-canonical Wnt effector driving Rac activation and migration, and C-terminal truncating mutations defined the PDZ-binding motif as essential for brain development.","evidence":"Reciprocal Co-IP, Rac assays, in vivo wound-healing knockout (signaling); homozygosity mapping and exome sequencing in hydrocephalus families (disease)","pmids":["22643886","23042809"],"confidence":"High","gaps":["Genetic disease evidence lacked direct functional assay of mutant protein","How Dvl association is coupled to Rac GEF machinery not fully resolved"]},{"year":2014,"claim":"A missense mutation showed DAPLE can engage stress signaling, connecting it to JNK activation and apoptosis in cerebellar neurodegeneration.","evidence":"Whole-exome sequencing plus cell-based JNK, caspase-3 cleavage, and apoptosis assays of mutant CCDC88C","pmids":["25062847"],"confidence":"Medium","gaps":["Single-lab cell-based readouts","Molecular mechanism linking R464H to JNK activation not defined"]},{"year":2015,"claim":"Defining the GBA motif resolved how DAPLE transduces Wnt to G proteins, showing it directly activates Gαi and binds Frizzled to suppress β-catenin while enhancing PI3K-Akt/Rac1 and invasiveness; gastric cancer work positioned it downstream of Wnt5a/Dvl for Rac/JNK.","evidence":"In vitro GEF assays, direct binding, GBA mutagenesis, signaling and invasion models; siRNA knockdown with Rac/JNK readouts and xenograft metastasis","pmids":["26126266","26577606"],"confidence":"High","gaps":["Structural basis of Frizzled-DAPLE binding not resolved","How GBA Gαi activation is spatially coordinated with receptor engagement unclear"]},{"year":2017,"claim":"In vivo and trafficking studies established DAPLE's roles in planar polarity of motile cilia and in Akt-regulated endosomal trafficking of β-catenin/E-cadherin.","evidence":"Daple knockout mouse cilia orientation and CSF flow analysis; phosphorylation, endosomal fractionation, and PI-binding/Akt-site mutagenesis; hair-cell PCP genetics","pmids":["28746879","29021338","29229865"],"confidence":"Medium","gaps":["Akt-trafficking mechanism shown in single lab","Link between cilia polarity defects and Wnt/G-protein signaling not mechanistically closed"]},{"year":2018,"claim":"Phosphoproteomics established that tyrosine kinases regulate DAPLE by phosphorylating sites near the Dvl-binding motif, dissociating the Dvl complex and augmenting Gαi activation and EMT.","evidence":"Mass spectrometry, in vitro kinase assays, Co-IP, Gαi activation assays, EMT assays","pmids":["29487190"],"confidence":"High","gaps":["Which kinases dominate in specific tissue contexts unclear","Quantitative relationship between phosphorylation and GEF output not defined"]},{"year":2019,"claim":"Domain-resolved binding studies and isoform analysis clarified that DAPLE docks at apical junctions through its PBM-MPDZ interaction for apical constriction, and that GBA-dependent tumor suppression is shared by isoforms while only full-length drives invasion.","evidence":"PDZ pulldowns, Co-IP, Xenopus morpholino constriction assays; isoform cloning with GBA mutagenesis and invasion assays","pmids":["31268831","31431650"],"confidence":"High","gaps":["Isoform-specific structural difference driving invasion not mapped","Single-lab Xenopus constriction data"]},{"year":2020,"claim":"A cluster of studies refined the GBA's pan-Gα selectivity, the junctional PARD3 docking and its tyrosine-phosphorylation control, the CK1ε-Dvl2 phosphorylation route to β-catenin, and TRIM11-mediated autophagic turnover.","evidence":"In vitro nucleotide exchange with M1669 mutagenesis; PDZ pulldowns and Src phosphorylation; trimeric Co-IP with Dvl2/CK1ε and reporter assays; ubiquitination and selective autophagy assays","pmids":["31949046","32058970","32888647","32382014"],"confidence":"Medium","gaps":["Several mechanisms from single labs without reciprocal validation","How opposing Gαi vs Gαs/Gαq effects integrate in cells unresolved"]},{"year":2022,"claim":"The apical actomyosin work integrated DAPLE into the PAR polarity complex, showing it is recruited by PAR3-PAR6-aPKC and assembles actomyosin via CD2AP recruitment and GPCR-independent RhoA activation.","evidence":"Reciprocal Co-IP, immunofluorescence, knockdown/overexpression, Rho activation and actomyosin assembly assays","pmids":["35389423"],"confidence":"High","gaps":["In vivo relevance of CD2AP recruitment not tested","Coupling between PAR recruitment and which Gα is activated locally unclear"]},{"year":2024,"claim":"Glycosylation control and a metastasis transcriptional axis were defined, showing GALNT6-mediated O-GalNAc glycosylation stabilizes DAPLE to support c-JUN-driven CEMIP expression and breast cancer metastasis.","evidence":"Co-IP, O-glycosylation assays, GALNT6 and CCDC88C gain/loss-of-function, c-JUN reporter, in vivo metastasis model","pmids":["38971758"],"confidence":"Medium","gaps":["Glycosylation sites on DAPLE not mapped","Single-lab in vivo metastasis evidence"]},{"year":2025,"claim":"Structural/biophysical and fusion-oncoprotein studies clarified that DAPLE's extended PBM binds DVL2 PDZ with exceptionally high affinity to mediate Wnt5a-induced primary cilia disassembly, and that its coiled-coil supports localization and maximal activation of CCDC88C-FLT3 fusions.","evidence":"Proximity labeling with structural/biophysical binding and cilia assays; cell-based FLT3 fusion kinase, STAT5/AKT/MAPK, and localization assays (both preprints)","pmids":["41256517"],"confidence":"Medium","gaps":["Both findings are preprints from single labs","Atomic-resolution structure of the DAPLE-DVL2 PDZ complex not finalized in peer-reviewed form"]},{"year":null,"claim":"How DAPLE's opposing actions on different Gα subunits, its multiple PDZ docking partners, and its many post-translational regulators are integrated into context-specific outputs in vivo remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model reconciling Gαi activation vs Gαs/Gαq inhibition in a physiological context","Tissue-specific deployment of canonical vs non-canonical Wnt outputs not mapped","Full-length structural organization of the scaffold unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,8]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,8,4]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[5,11,12]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[7]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[10,11,12]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,7]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[7]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[2,6,17]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[2,19]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[16]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,8]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,5,6]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[15,21]}],"complexes":["DAPLE-Dvl2-CK1ε trimeric complex","PAR3-PAR6-aPKC (PAR polarity) complex"],"partners":["DVL2","PARD3","MPDZ","CD2AP","CK1Ε","TRIM11","GALNT6","GNAI"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9P219","full_name":"Protein Daple","aliases":["Coiled-coil domain-containing protein 88C","Dvl-associating protein with a high frequency of leucine residues","hDaple","Hook-related protein 2","HkRP2"],"length_aa":2028,"mass_kda":228.2,"function":"Required for activation of guanine nucleotide-binding proteins (G-proteins) during non-canonical Wnt signaling (PubMed:26126266). Binds to ligand-activated Wnt receptor FZD7, displacing DVL1 from the FZD7 receptor and leading to inhibition of canonical Wnt signaling (PubMed:26126266). Acts as a non-receptor guanine nucleotide exchange factor by also binding to guanine nucleotide-binding protein G(i) alpha (Gi-alpha) subunits, leading to their activation (PubMed:26126266). Binding to Gi-alpha subunits displaces the beta and gamma subunits from the heterotrimeric G-protein complex, triggering non-canonical Wnt responses such as activation of RAC1 and PI3K-AKT signaling (PubMed:26126266). Promotes apical constriction of cells via ARHGEF18 (PubMed:30948426)","subcellular_location":"Cytoplasm; Cell junction","url":"https://www.uniprot.org/uniprotkb/Q9P219/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CCDC88C","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CCDC88C","total_profiled":1310},"omim":[{"mim_id":"620242","title":"NEURODEVELOPMENTAL DISORDER WITH POOR GROWTH AND BEHAVIORAL ABNORMALITIES; NEDGBA","url":"https://www.omim.org/entry/620242"},{"mim_id":"616053","title":"SPINOCEREBELLAR ATAXIA 40; SCA40","url":"https://www.omim.org/entry/616053"},{"mim_id":"611204","title":"COILED-COIL DOMAIN-CONTAINING PROTEIN 88C; CCDC88C","url":"https://www.omim.org/entry/611204"},{"mim_id":"609736","title":"COILED-COIL DOMAIN-CONTAINING PROTEIN 88A; CCDC88A","url":"https://www.omim.org/entry/609736"},{"mim_id":"609126","title":"ATPase, CLASS II, TYPE 9A; ATP9A","url":"https://www.omim.org/entry/609126"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Cell Junctions","reliability":"Enhanced"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":37.7}],"url":"https://www.proteinatlas.org/search/CCDC88C"},"hgnc":{"alias_symbol":["DAPLE","HkRP2","SCA40"],"prev_symbol":["KIAA1509"]},"alphafold":{"accession":"Q9P219","domains":[{"cath_id":"1.10.418.10","chopping":"6-175","consensus_level":"high","plddt":87.0833,"start":6,"end":175}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P219","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P219-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P219-F1-predicted_aligned_error_v6.png","plddt_mean":64.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CCDC88C","jax_strain_url":"https://www.jax.org/strain/search?query=CCDC88C"},"sequence":{"accession":"Q9P219","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9P219.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9P219/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P219"}},"corpus_meta":[{"pmid":"26126266","id":"PMC_26126266","title":"Daple is a novel non-receptor GEF required for trimeric G protein activation in Wnt signaling.","date":"2015","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/26126266","citation_count":100,"is_preprint":false},{"pmid":"21031079","id":"PMC_21031079","title":"Disturbed Wnt Signalling due to a Mutation in CCDC88C Causes an Autosomal Recessive Non-Syndromic Hydrocephalus with Medial Diverticulum.","date":"2010","source":"Molecular syndromology","url":"https://pubmed.ncbi.nlm.nih.gov/21031079","citation_count":75,"is_preprint":false},{"pmid":"32382014","id":"PMC_32382014","title":"TRIM11 facilitates chemoresistance in nasopharyngeal carcinoma by activating the β-catenin/ABCC9 axis via p62-selective autophagic degradation of Daple.","date":"2020","source":"Oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/32382014","citation_count":74,"is_preprint":false},{"pmid":"22643886","id":"PMC_22643886","title":"The Dishevelled-associating protein Daple controls the non-canonical Wnt/Rac pathway and cell motility.","date":"2012","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/22643886","citation_count":66,"is_preprint":false},{"pmid":"23042809","id":"PMC_23042809","title":"Two novel CCDC88C mutations confirm the role of DAPLE in autosomal recessive congenital hydrocephalus.","date":"2012","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23042809","citation_count":62,"is_preprint":false},{"pmid":"28746879","id":"PMC_28746879","title":"Daple Coordinates Planar Polarized Microtubule Dynamics in Ependymal Cells and Contributes to Hydrocephalus.","date":"2017","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/28746879","citation_count":60,"is_preprint":false},{"pmid":"25062847","id":"PMC_25062847","title":"A novel missense mutation in CCDC88C activates the JNK pathway and causes a dominant form of spinocerebellar ataxia.","date":"2014","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25062847","citation_count":54,"is_preprint":false},{"pmid":"15496975","id":"PMC_15496975","title":"KIAA1509 is a novel PDGFRB fusion partner in imatinib-responsive myeloproliferative disease associated with a t(5;14)(q33;q32).","date":"2005","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/15496975","citation_count":46,"is_preprint":false},{"pmid":"29229865","id":"PMC_29229865","title":"Daple coordinates organ-wide and cell-intrinsic polarity to pattern inner-ear hair bundles.","date":"2017","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/29229865","citation_count":35,"is_preprint":false},{"pmid":"26577606","id":"PMC_26577606","title":"Role for Daple in non-canonical Wnt signaling during gastric cancer invasion and metastasis.","date":"2015","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/26577606","citation_count":34,"is_preprint":false},{"pmid":"29341397","id":"PMC_29341397","title":"Bi-allelic mutations of CCDC88C are a rare cause of severe congenital hydrocephalus.","date":"2018","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/29341397","citation_count":31,"is_preprint":false},{"pmid":"16026968","id":"PMC_16026968","title":"Novel Daple-like protein positively regulates both the Wnt/beta-catenin pathway and the Wnt/JNK pathway in Xenopus.","date":"2005","source":"Mechanisms of development","url":"https://pubmed.ncbi.nlm.nih.gov/16026968","citation_count":24,"is_preprint":false},{"pmid":"29487190","id":"PMC_29487190","title":"Convergence of Wnt, growth factor, and heterotrimeric G protein signals on the guanine nucleotide exchange factor Daple.","date":"2018","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/29487190","citation_count":23,"is_preprint":false},{"pmid":"31268831","id":"PMC_31268831","title":"DAPLE and MPDZ bind to each other and cooperate to promote apical cell constriction.","date":"2019","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/31268831","citation_count":21,"is_preprint":false},{"pmid":"34092257","id":"PMC_34092257","title":"Neuropathological hallmarks of fetal hydrocephalus linked to CCDC88C pathogenic variants.","date":"2021","source":"Acta neuropathologica communications","url":"https://pubmed.ncbi.nlm.nih.gov/34092257","citation_count":21,"is_preprint":false},{"pmid":"24772479","id":"PMC_24772479","title":"Identification and functional characterization of imatinib-sensitive DTD1-PDGFRB and CCDC88C-PDGFRB fusion genes in eosinophilia-associated myeloid/lymphoid neoplasms.","date":"2014","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/24772479","citation_count":20,"is_preprint":false},{"pmid":"31949046","id":"PMC_31949046","title":"DAPLE protein inhibits nucleotide exchange on Gαs and Gαq via the same motif that activates Gαi.","date":"2020","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31949046","citation_count":14,"is_preprint":false},{"pmid":"29021338","id":"PMC_29021338","title":"A Daple-Akt feed-forward loop enhances noncanonical Wnt signals by compartmentalizing β-catenin.","date":"2017","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/29021338","citation_count":14,"is_preprint":false},{"pmid":"32888647","id":"PMC_32888647","title":"The Daple-CK1ε complex regulates Dvl2 phosphorylation and canonical Wnt signaling.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32888647","citation_count":12,"is_preprint":false},{"pmid":"33602173","id":"PMC_33602173","title":"A heterozygous mutation in the CCDC88C gene likely causes early-onset pure hereditary spastic paraplegia: a case report.","date":"2021","source":"BMC neurology","url":"https://pubmed.ncbi.nlm.nih.gov/33602173","citation_count":12,"is_preprint":false},{"pmid":"29225145","id":"PMC_29225145","title":"Surprisingly good outcome in antenatal diagnosis of severe hydrocephalus related to CCDC88C deficiency.","date":"2017","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29225145","citation_count":11,"is_preprint":false},{"pmid":"31431650","id":"PMC_31431650","title":"Two Isoforms of the Guanine Nucleotide Exchange Factor, Daple/CCDC88C Cooperate as Tumor Suppressors.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31431650","citation_count":10,"is_preprint":false},{"pmid":"30726835","id":"PMC_30726835","title":"Sustained Response to Imatinib in a Pediatric Patient with Concurrent Myeloproliferative Disease and Lymphoblastic Lymphoma Associated with a CCDC88C-PDGFRB Fusion Gene.","date":"2019","source":"Acta haematologica","url":"https://pubmed.ncbi.nlm.nih.gov/30726835","citation_count":9,"is_preprint":false},{"pmid":"35389423","id":"PMC_35389423","title":"DAPLE orchestrates apical actomyosin assembly from junctional polarity complexes.","date":"2022","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/35389423","citation_count":8,"is_preprint":false},{"pmid":"32058970","id":"PMC_32058970","title":"Tyrosine-Based Signals Regulate the Assembly of Daple⋅PARD3 Complex at Cell-Cell Junctions.","date":"2020","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/32058970","citation_count":8,"is_preprint":false},{"pmid":"34642354","id":"PMC_34642354","title":"Daple deficiency causes hearing loss in adult mice by inducing defects in cochlear stereocilia and apical microtubules.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/34642354","citation_count":6,"is_preprint":false},{"pmid":"38971758","id":"PMC_38971758","title":"CCDC88C, an O-GalNAc glycosylation substrate of GALNT6, drives breast cancer metastasis by promoting c-JUN-mediated CEMIP transcription.","date":"2024","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/38971758","citation_count":5,"is_preprint":false},{"pmid":"17681599","id":"PMC_17681599","title":"Extramedullary molecular evidence of the 5'KIAA1509/3'PDGFRB fusion gene in chronic eosinophilic leukemia.","date":"2007","source":"Leukemia research","url":"https://pubmed.ncbi.nlm.nih.gov/17681599","citation_count":5,"is_preprint":false},{"pmid":"32951102","id":"PMC_32951102","title":"Beneficial tyrosine kinase inhibitor therapy in a patient with relapsed BCR-ABL1-like acute lymphoblastic leukemia with CCDC88C-PDGFRB fusion.","date":"2020","source":"International journal of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/32951102","citation_count":5,"is_preprint":false},{"pmid":"36768938","id":"PMC_36768938","title":"Spinocerebellar Ataxia in a Hungarian Female Patient with a Novel Variant of Unknown Significance in the CCDC88C Gene.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36768938","citation_count":4,"is_preprint":false},{"pmid":"36704135","id":"PMC_36704135","title":"Case Report: Whole genome sequencing identifies CCDC88C as a novel JAK2 fusion partner in pediatric T-cell acute lymphoblastic leukemia.","date":"2023","source":"Frontiers in pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/36704135","citation_count":3,"is_preprint":false},{"pmid":"21054467","id":"PMC_21054467","title":"A case of myeloid neoplasm associated with eosinophilia and KIAA1509-PDGFRβ responsive to combination treatment with imatinib mesylate and prednisolone.","date":"2010","source":"Journal of clinical pharmacy and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/21054467","citation_count":1,"is_preprint":false},{"pmid":"38173219","id":"PMC_38173219","title":"CCDC88C variants are associated with focal epilepsy and genotype-phenotype correlation.","date":"2024","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/38173219","citation_count":0,"is_preprint":false},{"pmid":"41256517","id":"PMC_41256517","title":"Daple-FLT3 (CCDC88C-FLT3) gene fusion requires the coiled-coil domain for maximal activation and pericentrosomal localization.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/41256517","citation_count":0,"is_preprint":false},{"pmid":"37899026","id":"PMC_37899026","title":"A Novel c.3636-4 A>G Mutation in the CCDC88C Plays a Causative Role in Familial Spinocerebellar Ataxia.","date":"2023","source":"Human heredity","url":"https://pubmed.ncbi.nlm.nih.gov/37899026","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.26.678488","title":"Dishevelled drives Wnt-stimulated disassembly of primary cilia through a unique PDZ-mediated binding mode with Daple and Girdin","date":"2025-09-26","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.26.678488","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":19275,"output_tokens":5984,"usd":0.073792,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14710,"output_tokens":4894,"usd":0.09795,"stage2_stop_reason":"end_turn"},"total_usd":0.171742,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"DAPLE contains a Gα-binding and activating (GBA) motif that directly activates Gαi proteins, and an adjacent domain that directly binds Frizzled receptors (FZDRs), thereby linking Wnt stimulation to G protein activation. This triggers non-canonical Wnt responses: suppression of the β-catenin/TCF/LEF pathway and tumorigenesis, but enhancement of PI3K-Akt and Rac1 signals and tumor cell invasiveness.\",\n      \"method\": \"Biochemical binding assays, GEF activity assays, cell-based signaling assays, mutagenesis, tumor invasion models\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods including in vitro GEF assays, direct binding studies, mutagenesis of the GBA motif, and functional cellular readouts; foundational paper replicated by subsequent studies\",\n      \"pmids\": [\"26126266\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"DAPLE interacts directly with Dishevelled (Dvl) and regulates Wnt5a-mediated activation of Rac and formation of lamellipodia. DAPLE increases the association of Dvl with an isoform of atypical protein kinase C (aPKC/PKCλ), promoting Rac activation, and is required for fibroblast and epithelial cell migration during wound healing in vivo.\",\n      \"method\": \"Co-immunoprecipitation, lamellipodia formation assays, Rac activation assays, siRNA knockdown, in vivo wound healing model in mice\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in vivo knockout phenotype, Rac activation assays, multiple cell types tested; replicated in subsequent studies\",\n      \"pmids\": [\"22643886\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DAPLE-deficient mice develop hydrocephalus with ependymal cilia lacking coordinated orientation. DAPLE regulates microtubule dynamics at the anterior side of ependymal cells, orienting cilial basal bodies required for directional cerebrospinal fluid (CSF) flow, establishing a role for DAPLE in planar polarity of motile cilia.\",\n      \"method\": \"Daple-deficient (knockout) mouse model, cilia orientation analysis, microtubule dynamics imaging, CSF flow assay\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with defined cellular phenotype (cilia disorientation, hydrocephalus), direct imaging of microtubule dynamics; replicated across multiple labs in different contexts\",\n      \"pmids\": [\"28746879\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"A missense mutation in CCDC88C (p.R464H) activates the c-Jun N-terminal kinase (JNK) pathway, induces caspase-3 cleavage, and triggers apoptosis, linking DAPLE to JNK stress signaling and cerebellar neurodegeneration.\",\n      \"method\": \"Whole-exome sequencing, expression of mutant CCDC88C in cells, JNK pathway activation assay, caspase-3 cleavage assay, apoptosis assay\",\n      \"journal\": \"Journal of medical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-based functional assays with mutant protein and multiple readouts (JNK, caspase-3, apoptosis) in a single lab\",\n      \"pmids\": [\"25062847\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DAPLE is a substrate of multiple receptor tyrosine kinases (RTKs) and non-RTKs. Phosphorylation near the Dvl-binding motif causes DAPLE/Dvl complex dissociation and augments DAPLE's ability to bind to and activate Gαi, potentiating β-catenin-independent Wnt signals and stimulating epithelial-mesenchymal transition (EMT).\",\n      \"method\": \"Phosphoproteomics (mass spectrometry), Co-immunoprecipitation, in vitro kinase assay, Gαi activation assay, EMT assay\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — mass spectrometry identification of phosphorylation sites, in vitro kinase assays, Co-IP, Gαi nucleotide exchange assay, multiple orthogonal methods\",\n      \"pmids\": [\"29487190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DAPLE directly binds the PDZ3 domain of MPDZ via its PDZ-binding motif (PBM) and colocalizes with MPDZ at apical cell junctions. MPDZ depletion blunts DAPLE-mediated apical constriction of cultured cells. Both proteins are required for apical constriction of neuroepithelial cells and neural plate bending during Xenopus neurulation.\",\n      \"method\": \"Co-immunoprecipitation, direct binding assay (PDZ domain pulldown), colocalization imaging, Xenopus morpholino knockdown, cell constriction assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct binding demonstrated between defined domains, functional rescue in Xenopus, cellular apical constriction assay, multiple orthogonal methods\",\n      \"pmids\": [\"31268831\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DAPLE interacts with both PCP (planar cell polarity) and cell-intrinsic G-protein-based signals in inner-ear hair cells. DAPLE is required to maintain the polarized distribution of the core PCP protein Dishevelled and to position the primary cilium at the abneural edge of the apical surface, integrating organ-wide and cell-intrinsic polarity.\",\n      \"method\": \"Mouse knockout/mutant analysis, immunofluorescence localization of PCP proteins, cilium positioning assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with defined cellular phenotype (Dvl mislocalization, cilium mispositioning), single lab\",\n      \"pmids\": [\"29229865\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Akt phosphorylates the phosphoinositide (PI)-binding domain of DAPLE, abolishing DAPLE's ability to bind PI3-P-enriched endosomes and engage dynein motor complex for trafficking of β-catenin/E-cadherin complexes to pericentriolar recycling endosomes (PCREs). Phosphorylation compartmentalizes DAPLE/β-catenin/E-cadherin complexes to cell-cell contact sites, enhancing non-canonical Wnt signals and suppressing colony growth.\",\n      \"method\": \"Phosphorylation assay, endosomal fractionation, Co-immunoprecipitation, colony growth assay, mutagenesis of PI-binding domain and Akt phosphorylation site\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct phosphorylation assay, fractionation, mutant analysis, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"29021338\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DAPLE's GBA motif binds efficiently to Gαs and Gαq (in addition to Gαi) and inhibits nucleotide exchange on Gαs and Gαq, in contrast to the nucleotide-exchange acceleration it causes for Gαi. Met-1669 in the GBA motif was identified as the residue that enables better binding to Gαs and Gαq.\",\n      \"method\": \"In vitro nucleotide exchange assay, binding assay, site-directed mutagenesis (M1669 substitution)\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical reconstitution with mutagenesis defining mechanism; single lab but with rigorous in vitro assays\",\n      \"pmids\": [\"31949046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TRIM11 associates with DAPLE and promotes its ubiquitin-mediated degradation via a p62-selective autophagic pathway, thereby upregulating β-catenin and inducing ABCC9 expression in nasopharyngeal carcinoma drug-resistant cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, selective autophagy assay, β-catenin reporter assay\",\n      \"journal\": \"Oncogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP showing TRIM11–DAPLE interaction, ubiquitination assay, autophagic degradation shown, single lab with multiple assays\",\n      \"pmids\": [\"32382014\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DAPLE forms a trimeric complex with Dvl2 and CK1ε. Daple overexpression induces CK1ε-mediated phosphorylation of Dvl2 at Thr224, which is required for full activation of β-catenin transcriptional activity. A Daple mutant lacking the C-terminal GCV motif (Dvl-binding) retains CK1ε interaction but fails to induce Dvl phosphorylation. Wnt3a stimulation increases membrane localization of DAPLE and its association with Dvl.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (ΔGCV, Dvl2-T224A), β-catenin reporter assay, membrane fractionation\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, mutagenesis of interaction domains, functional reporter assay, single lab\",\n      \"pmids\": [\"32888647\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DAPLE localizes to epithelial cell junctions via its PDZ-binding motif (PBM) binding the third PDZ domain of PARD3. Tyrosine phosphorylation of DAPLE's PBM by receptor and non-receptor tyrosine kinases (including Src) regulates the interaction: hypophosphorylation strengthens DAPLE–PARD3 interaction, while hyperphosphorylation disrupts it, controlling contact-triggered planar polarity.\",\n      \"method\": \"Co-immunoprecipitation, direct binding assay (PDZ domain pulldown), tyrosine kinase assay (Src), immunofluorescence localization, phosphomimetic/phosphonull mutagenesis\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct PDZ binding assay, phosphorylation by Src demonstrated, PBM mutagenesis, multiple methods in single lab\",\n      \"pmids\": [\"32058970\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The PAR polarity complex (PAR3-PAR6-aPKC) at apical cell junctions recruits DAPLE, which in turn triggers a two-pronged mechanism for assembly of the apical actomyosin network: (1) direct recruitment of the actin-stabilizing protein CD2AP to apical junctions, and (2) GPCR-independent activation of heterotrimeric G protein signaling to favor RhoA-myosin activation.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, DAPLE knockdown/overexpression, Rho activation assay, actomyosin assembly assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, functional knockdown with defined actomyosin phenotype, G protein activation assay, multiple orthogonal methods in one study\",\n      \"pmids\": [\"35389423\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DAPLE (CCDC88C) is a substrate of GALNT6, which promotes O-linked GalNAc glycosylation of DAPLE, maintaining its protein stability. This O-GalNAc modification is required for DAPLE's pro-metastatic function. DAPLE promotes breast cancer metastasis via c-JUN-mediated transcriptional activation of CEMIP.\",\n      \"method\": \"Co-immunoprecipitation, O-glycosylation assay, GALNT6 knockdown/overexpression, CCDC88C gain/loss-of-function, c-JUN reporter assay, in vivo metastasis model\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — O-glycosylation biochemically demonstrated, functional stability assay, in vivo metastasis model, single lab\",\n      \"pmids\": [\"38971758\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DAPLE depletion suppresses Wnt5a-induced Rac and JNK activation, laminin γ2 expression, and cell migration/invasion in gastric cancer cells. Daple depletion also suppressed liver metastasis in a mouse xenograft model, positioning DAPLE downstream of Wnt5a/Dvl to activate Rac and JNK in gastric cancer.\",\n      \"method\": \"siRNA knockdown, Rac activation assay, JNK phosphorylation assay, laminin γ2 expression assay, Transwell migration/invasion assay, mouse xenograft metastasis model\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with multiple defined pathway readouts and in vivo validation, single lab\",\n      \"pmids\": [\"26577606\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"CCDC88C (KIAA1509) contains a coiled-coil oligomerization domain with homology to the HOOK family of proteins. The KIAA1509-PDGFRβ fusion protein resulting from t(5;14)(q33;q32) retains the KIAA1509 coiled-coil domain fused to the cytoplasmic kinase domain of PDGFRβ, producing a constitutively active tyrosine kinase that transforms hematopoietic cells.\",\n      \"method\": \"Inverse PCR cloning, sequencing, imatinib treatment (clinical response), domain analysis\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fusion gene cloned and characterized; imatinib clinical response confirms constitutive kinase activity; replicated in multiple subsequent cases\",\n      \"pmids\": [\"15496975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The CCDC88C-FLT3 (Daple-FLT3) fusion oncoprotein contains a constitutively active FLT3 kinase domain that activates STAT5a, AKT, and MAPK signaling without ligand stimulation. The coiled-coil domain of DAPLE is dispensable for kinase activation but necessary for maximal activation and pericentrosomal localization of the fusion protein. Sorafenib (and to a lesser degree imatinib) can modulate this signaling.\",\n      \"method\": \"Cell-based kinase activity assay, STAT5/AKT/MAPK phosphorylation assay, subcellular localization imaging, coiled-coil domain deletion mutagenesis, TKI treatment\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-based functional assay with domain mutagenesis and localization imaging; preprint, single lab\",\n      \"pmids\": [\"41256517\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DAPLE and its paralog Girdin-L bear unique extended C-terminal PDZ-binding motifs that bind to the PDZ domain of DVL2 with exceptionally high affinity, as revealed by proximity labeling combined with structural and biophysical analysis. Deletion of these motifs or the DVL2 PDZ domain results in elongated primary cilia and renders cilia unresponsive to Wnt5a-stimulated disassembly, establishing that the DVL2 PDZ–DAPLE interaction mediates primary cilia disassembly in response to Wnt5a.\",\n      \"method\": \"Proximity labeling (BioID), structural analysis, biophysical binding assay, PDZ motif deletion mutagenesis, primary cilia length measurement, Wnt5a stimulation assay in HEK293T cells\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — structural and biophysical binding with mutagenesis, functional cilia assay; preprint, single lab, not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.09.26.678488\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Two isoforms of DAPLE/CCDC88C both suppress tumor growth via their GBA motif, but only the full-length isoform triggers EMT and invasion. Both isoforms are suppressed during colon cancer progression, and their reduced expression carries additive prognostic significance.\",\n      \"method\": \"Isoform cloning, GBA motif mutagenesis, colony formation assay, invasion assay\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional mutagenesis of GBA motif, isoform-specific gain-of-function, multiple cellular readouts, single lab\",\n      \"pmids\": [\"31431650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Adult Daple-knockout mice exhibit hearing disturbances across a broad frequency range. In these mice, apical microtubules in cochlear hair cells are irregularly aggregated and the number of microtubules attached to plasma membranes is decreased, causing distorted hair bundle arrangement. Nocodazole treatment in wild-type cochlea culture phenocopied these microtubule defects, establishing DAPLE's role in apical microtubule organization and hair bundle patterning.\",\n      \"method\": \"Daple knockout mouse audiometry (auditory brainstem response), immunofluorescence of hair bundles, electron/confocal microscopy of apical microtubules, nocodazole pharmacological experiment\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout with defined audiological and cellular phenotype, pharmacological validation, single lab\",\n      \"pmids\": [\"34642354\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"A homozygous splice-site mutation in CCDC88C (intron 29) causes loss of DAPLE protein. In blood from the affected patient, CTNNB1 (β-catenin) and LEF1 mRNA levels were increased compared to controls, consistent with loss of DAPLE's inhibitory role on canonical Wnt/β-catenin signaling.\",\n      \"method\": \"Western blotting (protein loss), quantitative RT-PCR of Wnt pathway genes, positional cloning\",\n      \"journal\": \"Molecular syndromology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single family, mRNA expression changes only (no direct pathway manipulation), no rescue experiment\",\n      \"pmids\": [\"21031079\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Mutations truncating the extreme C-terminus of DAPLE, including the GCV PDZ-binding motif, cause autosomal recessive congenital hydrocephalus, indicating that the PDZ domain-binding motif (which mediates Dishevelled binding) is functionally essential for DAPLE's role in brain development.\",\n      \"method\": \"Homozygosity mapping, whole exome sequencing, protein domain analysis of truncating mutations\",\n      \"journal\": \"Journal of medical genetics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — genetic evidence from human families; no direct in vitro functional assay; mechanistic inference from domain knowledge\",\n      \"pmids\": [\"23042809\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DAPLE/CCDC88C is a multi-domain cytoplasmic scaffold and non-receptor guanine nucleotide exchange factor (GEF) that bridges Wnt/Frizzled receptor signaling to heterotrimeric G protein activation: its GBA motif activates Gαi while inhibiting Gαs and Gαq, its C-terminal PDZ-binding motif (GCV) docks to the PDZ domain of Dishevelled (DVL2) and PARD3/MPDZ at apical junctions, and it is regulated by phosphorylation from RTKs/Src (which dissociates the Dvl complex and augments Gαi activation) and by Akt (which controls endosomal trafficking of β-catenin); through these mechanisms DAPLE coordinates non-canonical Wnt-driven Rac activation and cell migration, apical actomyosin assembly via CD2AP recruitment and RhoA activation, planar polarity in ependymal and hair cells via apical microtubule organization, primary cilia disassembly in response to Wnt5a, and CK1ε-mediated Dvl2 phosphorylation that activates canonical β-catenin signaling, while its dysregulation—through loss-of-function mutations, gain-of-function JNK-activating mutations, oncogenic CCDC88C-PDGFRB/FLT3/JAK2 chromosomal fusions, or TRIM11-mediated autophagic degradation—underlies congenital hydrocephalus, spinocerebellar ataxia, and cancer progression.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CCDC88C/DAPLE is a multi-domain cytoplasmic scaffold that couples Wnt/Frizzled signaling to heterotrimeric G protein activation through a Gα-binding and activating (GBA) motif that accelerates nucleotide exchange on Gαi while binding and inhibiting nucleotide exchange on Gαs and Gαq, the latter selectivity conferred by Met-1669 [#0, #8]. By engaging Frizzled and Dishevelled, DAPLE drives non-canonical Wnt responses—suppressing β-catenin/TCF/LEF output while potentiating PI3K-Akt and Rac1 signaling, lamellipodia formation, and cell migration—and acts downstream of Wnt5a/Dvl to activate Rac and JNK [#0, #1, #14]. Its C-terminal PDZ-binding motif docks DAPLE at apical junctions through high-affinity interactions with the PDZ domains of DVL2, PARD3, and MPDZ, where it organizes epithelial polarity and apical actomyosin assembly: recruited by the PAR3-PAR6-aPKC complex, DAPLE both recruits the actin-stabilizing protein CD2AP and triggers GPCR-independent RhoA-myosin activation [#5, #11, #12]. DAPLE governs planar polarity and ciliary architecture, orienting ependymal cilia and positioning the primary cilium via apical microtubule organization [#2, #6, #19]. DAPLE activity is gated by post-translational regulation: tyrosine kinases including Src phosphorylate sites near the Dvl- and PARD3-binding motifs to dissociate these complexes and augment Gαi activation and EMT, Akt phosphorylates its phosphoinositide-binding domain to control endosomal trafficking of β-catenin/E-cadherin complexes, and protein levels are set by GALNT6-mediated O-GalNAc glycosylation and by TRIM11-driven selective autophagic degradation [#4, #7, #9, #13]. Human loss-of-function and C-terminal truncating mutations in CCDC88C cause autosomal recessive congenital hydrocephalus, and a p.R464H missense mutation that activates JNK and apoptosis is linked to cerebellar neurodegeneration [#21, #3]. Distinct from the full-length signaling protein, the CCDC88C coiled-coil oligomerization domain forms constitutively active oncogenic kinase fusions with PDGFRβ and FLT3 that transform hematopoietic cells [#15, #16].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Before its signaling role was known, CCDC88C was identified as a fusion partner whose coiled-coil domain could constitutively activate a kinase, establishing the first disease link via an oncogenic translocation.\",\n      \"evidence\": \"Inverse PCR cloning and sequencing of a t(5;14) KIAA1509-PDGFRβ fusion with imatinib clinical response\",\n      \"pmids\": [\"15496975\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not address the normal cellular function of full-length CCDC88C\", \"Mechanism of oligomerization-driven kinase activation not structurally resolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"A human splice-site mutation causing DAPLE protein loss first implicated the gene in restraining canonical Wnt output, raising elevated β-catenin and LEF1 transcripts.\",\n      \"evidence\": \"Positional cloning, Western blot of protein loss, and qRT-PCR of Wnt pathway genes in a single family\",\n      \"pmids\": [\"21031079\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single family with mRNA correlation only, no pathway manipulation or rescue\", \"Direct biochemical link to β-catenin regulation not established\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Linking DAPLE to Dishevelled established it as a non-canonical Wnt effector driving Rac activation and migration, and C-terminal truncating mutations defined the PDZ-binding motif as essential for brain development.\",\n      \"evidence\": \"Reciprocal Co-IP, Rac assays, in vivo wound-healing knockout (signaling); homozygosity mapping and exome sequencing in hydrocephalus families (disease)\",\n      \"pmids\": [\"22643886\", \"23042809\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genetic disease evidence lacked direct functional assay of mutant protein\", \"How Dvl association is coupled to Rac GEF machinery not fully resolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"A missense mutation showed DAPLE can engage stress signaling, connecting it to JNK activation and apoptosis in cerebellar neurodegeneration.\",\n      \"evidence\": \"Whole-exome sequencing plus cell-based JNK, caspase-3 cleavage, and apoptosis assays of mutant CCDC88C\",\n      \"pmids\": [\"25062847\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab cell-based readouts\", \"Molecular mechanism linking R464H to JNK activation not defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defining the GBA motif resolved how DAPLE transduces Wnt to G proteins, showing it directly activates Gαi and binds Frizzled to suppress β-catenin while enhancing PI3K-Akt/Rac1 and invasiveness; gastric cancer work positioned it downstream of Wnt5a/Dvl for Rac/JNK.\",\n      \"evidence\": \"In vitro GEF assays, direct binding, GBA mutagenesis, signaling and invasion models; siRNA knockdown with Rac/JNK readouts and xenograft metastasis\",\n      \"pmids\": [\"26126266\", \"26577606\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Frizzled-DAPLE binding not resolved\", \"How GBA Gαi activation is spatially coordinated with receptor engagement unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"In vivo and trafficking studies established DAPLE's roles in planar polarity of motile cilia and in Akt-regulated endosomal trafficking of β-catenin/E-cadherin.\",\n      \"evidence\": \"Daple knockout mouse cilia orientation and CSF flow analysis; phosphorylation, endosomal fractionation, and PI-binding/Akt-site mutagenesis; hair-cell PCP genetics\",\n      \"pmids\": [\"28746879\", \"29021338\", \"29229865\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Akt-trafficking mechanism shown in single lab\", \"Link between cilia polarity defects and Wnt/G-protein signaling not mechanistically closed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Phosphoproteomics established that tyrosine kinases regulate DAPLE by phosphorylating sites near the Dvl-binding motif, dissociating the Dvl complex and augmenting Gαi activation and EMT.\",\n      \"evidence\": \"Mass spectrometry, in vitro kinase assays, Co-IP, Gαi activation assays, EMT assays\",\n      \"pmids\": [\"29487190\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which kinases dominate in specific tissue contexts unclear\", \"Quantitative relationship between phosphorylation and GEF output not defined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Domain-resolved binding studies and isoform analysis clarified that DAPLE docks at apical junctions through its PBM-MPDZ interaction for apical constriction, and that GBA-dependent tumor suppression is shared by isoforms while only full-length drives invasion.\",\n      \"evidence\": \"PDZ pulldowns, Co-IP, Xenopus morpholino constriction assays; isoform cloning with GBA mutagenesis and invasion assays\",\n      \"pmids\": [\"31268831\", \"31431650\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Isoform-specific structural difference driving invasion not mapped\", \"Single-lab Xenopus constriction data\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"A cluster of studies refined the GBA's pan-Gα selectivity, the junctional PARD3 docking and its tyrosine-phosphorylation control, the CK1ε-Dvl2 phosphorylation route to β-catenin, and TRIM11-mediated autophagic turnover.\",\n      \"evidence\": \"In vitro nucleotide exchange with M1669 mutagenesis; PDZ pulldowns and Src phosphorylation; trimeric Co-IP with Dvl2/CK1ε and reporter assays; ubiquitination and selective autophagy assays\",\n      \"pmids\": [\"31949046\", \"32058970\", \"32888647\", \"32382014\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Several mechanisms from single labs without reciprocal validation\", \"How opposing Gαi vs Gαs/Gαq effects integrate in cells unresolved\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"The apical actomyosin work integrated DAPLE into the PAR polarity complex, showing it is recruited by PAR3-PAR6-aPKC and assembles actomyosin via CD2AP recruitment and GPCR-independent RhoA activation.\",\n      \"evidence\": \"Reciprocal Co-IP, immunofluorescence, knockdown/overexpression, Rho activation and actomyosin assembly assays\",\n      \"pmids\": [\"35389423\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of CD2AP recruitment not tested\", \"Coupling between PAR recruitment and which Gα is activated locally unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Glycosylation control and a metastasis transcriptional axis were defined, showing GALNT6-mediated O-GalNAc glycosylation stabilizes DAPLE to support c-JUN-driven CEMIP expression and breast cancer metastasis.\",\n      \"evidence\": \"Co-IP, O-glycosylation assays, GALNT6 and CCDC88C gain/loss-of-function, c-JUN reporter, in vivo metastasis model\",\n      \"pmids\": [\"38971758\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Glycosylation sites on DAPLE not mapped\", \"Single-lab in vivo metastasis evidence\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Structural/biophysical and fusion-oncoprotein studies clarified that DAPLE's extended PBM binds DVL2 PDZ with exceptionally high affinity to mediate Wnt5a-induced primary cilia disassembly, and that its coiled-coil supports localization and maximal activation of CCDC88C-FLT3 fusions.\",\n      \"evidence\": \"Proximity labeling with structural/biophysical binding and cilia assays; cell-based FLT3 fusion kinase, STAT5/AKT/MAPK, and localization assays (both preprints)\",\n      \"pmids\": [\"41256517\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Both findings are preprints from single labs\", \"Atomic-resolution structure of the DAPLE-DVL2 PDZ complex not finalized in peer-reviewed form\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DAPLE's opposing actions on different Gα subunits, its multiple PDZ docking partners, and its many post-translational regulators are integrated into context-specific outputs in vivo remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model reconciling Gαi activation vs Gαs/Gαq inhibition in a physiological context\", \"Tissue-specific deployment of canonical vs non-canonical Wnt outputs not mapped\", \"Full-length structural organization of the scaffold unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 8]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 8, 4]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [5, 11, 12]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [10, 11, 12]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 7]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [2, 6, 17]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [2, 19]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [16]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 5, 6]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [15, 21]}\n    ],\n    \"complexes\": [\n      \"DAPLE-Dvl2-CK1\\u03b5 trimeric complex\",\n      \"PAR3-PAR6-aPKC (PAR polarity) complex\"\n    ],\n    \"partners\": [\n      \"DVL2\",\n      \"PARD3\",\n      \"MPDZ\",\n      \"CD2AP\",\n      \"CK1\\u03b5\",\n      \"TRIM11\",\n      \"GALNT6\",\n      \"GNAI\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":7,"faith_total":7,"faith_pct":100.0}}