{"gene":"CAMSAP3","run_date":"2026-06-09T22:57:17","timeline":{"discoveries":[{"year":2012,"finding":"CAMSAP3 (Nezha) and CAMSAP2 co-cluster at the minus ends of noncentrosomal microtubules in epithelial cells, stabilizing them. Depletion of both CAMSAPs caused marked reduction of microtubules with polymerizing plus ends and induced centrosomal microtubule growth, demonstrating that CAMSAP2 and CAMSAP3 cooperatively maintain noncentrosomal microtubules and suppress centrosomal microtubule-organizing activity.","method":"siRNA depletion, immunofluorescence, live imaging in epithelial cells","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with specific phenotypic readout replicated across two CAMSAP family members, independent validation of minus-end localization, replicated in subsequent studies","pmids":["23169647"],"is_preprint":false},{"year":2015,"finding":"CAMSAP3 accumulates at apical cortices of intestinal and Caco-2 epithelial cells and tethers longitudinal microtubule minus ends to these sites, thereby orienting microtubule arrays along the apico-basal axis with minus ends apical. The CC1 domain of CAMSAP3 is required for apical localization; forced mislocalization of CAMSAP3 disrupts epithelial architecture. Loss of CAMSAP3 causes random microtubule orientation and perturbed Golgi/nucleus positioning.","method":"Camsap3 mutation/siRNA knockdown, domain deletion analysis, forced mislocalization, immunofluorescence in mouse intestinal cells and Caco-2 cells","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function combined with domain mapping and forced mislocalization rescue, replicated in two cell systems","pmids":["26715742"],"is_preprint":false},{"year":2016,"finding":"ACF7 (MACF1), a spectraplakin, specifically binds to CAMSAP3 and is required for apical localization of CAMSAP3-decorated microtubule minus ends in intestinal epithelial cells. Knockout of CAMSAP3 (but not CAMSAP2) caused microtubule reorganization into a radial centrosomal array, redistribution of Rab11-positive endosomes from apical surface to pericentrosomal region, and inhibition of actin brush border formation at the apical side.","method":"Co-IP/binding assay, CAMSAP3 knockout, CAMSAP2 knockout, immunofluorescence, 3D cyst culture","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding demonstrated, KO with specific phenotypic readouts, two orthogonal KO comparisons, independently confirmed by a second lab (PMID 27693509)","pmids":["27802168"],"is_preprint":false},{"year":2016,"finding":"ACF7 interacts with CAMSAP3 at the minus ends of noncentrosomal microtubules and anchors them to actin filaments in Caco-2 epithelial cells. This CAMSAP3-ACF7 complex enables noncentrosomal microtubules to crosstalk with F-actin via retrograde flow, maintaining microtubule length and perpendicular orientation to the cell edge, and regulating focal adhesion size and cell migration.","method":"Co-IP, pulldown, live imaging, siRNA knockdown, focal adhesion/migration assays in Caco-2 cells","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP plus functional phenotypes (migration, focal adhesion), two orthogonal methods, consistent with parallel findings from another lab (PMID 27802168)","pmids":["27693509"],"is_preprint":false},{"year":2016,"finding":"CAMSAP3 binds CG-NAP (centrosome and Golgi localized PKN-associated protein) and plays a role in translocation and clustering of Golgi vesicles in epithelial cells. Depletion of either CAMSAP3 or CG-NAP causes Golgi membrane fragmentation. Stathmin-dependent microtubule dynamics, highest at the perinuclear region, also contributes to perinuclear Golgi distribution.","method":"Co-IP, siRNA knockdown, immunofluorescence in epithelial cells","journal":"Journal of genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP interaction with CG-NAP and loss-of-function phenotype, single lab, two methods","pmids":["28089391"],"is_preprint":false},{"year":2016,"finding":"CDH23-C (C isoform of cadherin-related 23) directly binds the CKK domain of CAMSAP3 via a conserved N-terminal CKK-binding motif (CBM), and this interaction inhibits CAMSAP3-induced microtubule bundle formation. The Usher Syndrome 1D-associated mutation CDH23 R3175H (mouse R55H) reduces CDH23-C interaction with CAMSAP3 both in vivo and in vitro.","method":"In vitro binding assay, pulldown, in vitro MT bundling assay, site-directed mutagenesis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with mutagenesis, single lab","pmids":["27349180"],"is_preprint":false},{"year":2017,"finding":"CAMSAP3 accumulates in the pericentrosomal area and accompanies microtubule release from the centrosome. Depletion of CAMSAP3 prevents MT release and causes MT focusing at centrosomes. CAMSAP3 coordinates with dynein and katanin to regulate MT detachment from the centrosome, identifying it as a key molecule for generation of noncentrosomal microtubules.","method":"siRNA knockdown, immunofluorescence, live imaging, co-depletion experiments with dynein and katanin in epithelial cells","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with specific subcellular phenotype, epistatic relationship with katanin/dynein established, single lab","pmids":["28386021"],"is_preprint":false},{"year":2018,"finding":"CAMSAP3 is enriched in axons of hippocampal neurons where it localizes preferentially along less-acetylated microtubules. CAMSAP3 mutation causes supernumerary axons and increased nocodazole-resistant/acetylated microtubules. CAMSAP3 depletion promotes tubulin acetylation and overexpression inhibits it, indicating CAMSAP3 retains a nonacetylated microtubule pool by interfering with αTAT1 activity. Depletion of αTAT1 abolished CAMSAP3-loss-dependent multiple-axon formation, placing CAMSAP3 upstream of αTAT1 in neuronal polarity maintenance.","method":"Camsap3 mouse mutation, siRNA knockdown, overexpression, αTAT1 depletion epistasis, immunofluorescence, nocodazole resistance assay in hippocampal neurons","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis (αTAT1 rescue), multiple loss-of-function approaches, quantitative phenotypic readouts, in vivo mutation","pmids":["30190432"],"is_preprint":false},{"year":2018,"finding":"CAMSAP3 loss in lung carcinoma cells increases tubulin acetylation, which upregulates Akt (protein kinase B) activity and promotes epithelial-to-mesenchymal transition (EMT) at the transcriptional level. CAMSAP3 therefore functions to suppress Akt activity via microtubule regulation, protecting epithelial phenotype.","method":"CAMSAP3 deletion, immunoblotting, EMT marker analysis, Akt activity assay in lung carcinoma cell lines","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with mechanistic pathway (acetylation→Akt→EMT), single lab, two orthogonal readouts","pmids":["30282632"],"is_preprint":false},{"year":2020,"finding":"CAMSAP3 localizes to the base of axonemes and at basal bodies in multiciliated cells (MCCs). In Camsap3 knockdown/hypomorph mice and Xenopus morpholino knockdown, the central microtubule pairs are missing from the majority of cilia, basal body polarity is disorganized, and synchronized ciliary beating is defective. This identifies CAMSAP3 as required for central MT pair assembly/stability in motile cilia.","method":"Camsap3 hypomorph mouse model, Xenopus morpholino knockdown, immunofluorescence, electron microscopy, ciliary beat analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — two independent model organisms (mouse and Xenopus), electron microscopy structural validation, loss-of-function with defined phenotype, replicated in subsequent studies (PMIDs 34319756, 35783105)","pmids":["32482850"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 concentrates at multiple sites of the cilium-basal body complex including the axonemal basal plate where the central pair of microtubules (CP) initiates. CAMSAP3 dysfunction results in loss of the CP, partial distortion of the basal plate, and failure of multicilia to undergo synchronized beating in tracheal airway epithelial cells.","method":"Camsap3 mutant mouse model, immunofluorescence, electron microscopy, high-speed videomicroscopy of ciliary beating","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with structural (EM) and functional (beat) readouts, consistent with independent findings (PMID 32482850)","pmids":["34319756"],"is_preprint":false},{"year":2021,"finding":"In mouse oviduct MCCs, CAMSAP3 localizes to the base of cilia in a polarized manner and is critical for intracellular basal body (BB) orientation and for assembly of microtubules interconnecting basal bodies. CAMSAP3 mutation disrupts intracellular coordination of BB orientation without affecting planar cell polarity (PCP) factor localization, placing CAMSAP3 function downstream of or parallel to PCP signaling.","method":"Camsap3 mutant mice, immunofluorescence, super-resolution microscopy, genetic comparison with CELSR1 mutants","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with structural readout, epistatic placement relative to PCP, single lab","pmids":["33468623"],"is_preprint":false},{"year":2021,"finding":"In ependymal cells, CAMSAP3 concentrates at apical domains to generate MT networks. Camsap3-mutated mice exhibit narrowed lateral ventricles due to failure of ependymal cells to broaden their apical domain. mTORC1 activity, required for ependymal cell growth, is downregulated in mutant cells, and lysosomes (which activate mTORC1) are reduced at apical regions where MT networks are disorganized, linking CAMSAP3-dependent MT organization to mTORC1 signaling and ventricle shaping.","method":"Camsap3 mutant mouse, immunofluorescence, lysosome localization assay, mTORC1 activity measurement, apical domain morphometry","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with pathway mechanistic follow-up (mTORC1/lysosome), single lab, two orthogonal mechanistic readouts","pmids":["33462112"],"is_preprint":false},{"year":2021,"finding":"In mouse proximal convoluted tubules, CAMSAP3 mutation perturbs microtubule arrays and activates mechanosensors YAP and PIEZO1, leading to enhanced cell proliferation, cell flattening, and cyst formation. This places CAMSAP3-mediated microtubule networks upstream of mechanosensitive YAP/PIEZO1 signaling in renal tubule morphology maintenance.","method":"Camsap3 mutant mice, immunofluorescence, YAP/PIEZO1 activity assays, histology","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with mechanistic pathway placement (YAP/PIEZO1), single lab","pmids":["33712686"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 knockout promotes lung cancer cell senescence-associated phenotypes and G1 cell cycle arrest. Mechanistically, phosphorylated ERK is markedly downregulated in CAMSAP3-deleted cells, suppressing cyclin D1. Proteomic analysis identified vimentin as a scaffold required for CAMSAP3-modulated ERK signaling. Full-length CAMSAP3 abrogates these phenotypes.","method":"CAMSAP3 CRISPR knockout, immunoprecipitation/mass spectrometry, Western blotting, xenograft model","journal":"Cancer medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO with proteomic identification of scaffold (vimentin), in vivo validation, single lab","pmids":["34724356"],"is_preprint":false},{"year":2023,"finding":"Paracingulin (CGNL1), but not PLEKHA7, recruits CAMSAP3 to tight junctions. GST pulldown shows CGNL1 interacts directly with CAMSAP3 via their respective coiled-coil regions. Knockout of CGNL1 causes loss of junctional CAMSAP3, disorganized cytoplasmic microtubules, irregular nuclei alignment, altered cyst morphogenesis, and disrupted planar apical microtubules. Ultrastructure expansion microscopy shows CAMSAP3-capped microtubules are tethered to junctions by the ZO-1-associated pool of CGNL1.","method":"GST pulldown, CGNL1/PLEKHA7 knockout, ultrastructure expansion microscopy, immunofluorescence, 3D cyst culture, in vivo mouse intestinal epithelium","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct biochemical interaction (GST pulldown) combined with KO phenotype and super-resolution structural imaging, two orthogonal methods, in vitro and in vivo validation","pmids":["37013686"],"is_preprint":false},{"year":2023,"finding":"CAMSAP3 interacts with nucleolin (NCL) to regulate HIF-1α mRNA stabilization. In CAMSAP3-knockout NSCLC cells, HIF-1α expression is upregulated along with VEGFA and MMPs 2 and 9, increasing invasion and angiogenesis. These effects were attenuated by reintroduction of wild-type CAMSAP3.","method":"CAMSAP3 CRISPR knockout, immunoprecipitation, mass spectrometry, RNA immunoprecipitation, qRT-PCR, in vivo xenograft","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interaction with NCL identified by MS, RNA-IP for mRNA stabilization, rescue experiment, single lab","pmids":["37019300"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 forms dimers through its C-terminal α-helix domain. This dimerization enhances the microtubule-binding affinity of the CKK domain and enables the CKK domain to regulate microtubule dynamics. The combined action of the microtubule-binding domain (MBD) and the C-terminal α-helix enables CAMSAP3 to decorate minus ends and achieve dynamic regulation of microtubule minus ends.","method":"Biochemical dimerization assays, domain deletion/mutagenesis, in vitro microtubule binding and dynamics assays","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with domain mutagenesis establishing dimerization mechanism and CKK domain enhancement, single lab but multiple orthogonal biochemical methods","pmids":["39479887"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 interacts with HMGB1, particularly with its acetylated form. In the presence of the HDAC inhibitor trichostatin A (TSA), CAMSAP3-HMGB1 complex formation is elevated, facilitating HMGB1 cytoplasmic translocation and secretion, thereby inducing autophagic cell death. CAMSAP3 knockout abolishes TSA-mediated autophagic cell death in lung cancer cells.","method":"CRISPR-Cas9 knockout, immunoprecipitation, proteomics, immunofluorescence, autophagic detection assays","journal":"Biochimica et biophysica acta. General subjects","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interaction with acetylated HMGB1 and KO functional phenotype, two orthogonal methods, single lab","pmids":["38598971"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 is essential for proper localization of the microtubule-dependent endosomal-lysosomal system. CAMSAP3 depletion causes EGFR translocation to the perinuclear MTOC, blocking plasma membrane recycling and promoting lysosomal EGFR degradation, thereby reducing EGFR signaling and increasing osimertinib resistance in NSCLC.","method":"CAMSAP3 siRNA knockdown and overexpression, immunofluorescence, in vitro and xenograft drug sensitivity assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function and gain-of-function with defined subcellular trafficking phenotype, in vivo xenograft validation, single lab","pmids":["41381443"],"is_preprint":false},{"year":2025,"finding":"Camsap3-mediated microtubules are present in transzonal projections (TZPs) of granulosa cells extending to oocytes in ovarian follicles. Camsap3 knockout mice are infertile and lack ovulation; their follicles show reduced TZP numbers, disorganized TZP microtubules, and decreased granulosa cell-oocyte contact, identifying CAMSAP3 as required for TZP architecture and follicle maturation.","method":"Camsap3 knockout mice, super-resolution microscopy, follicle counting, fertility assays","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with specific structural and functional phenotypes, super-resolution imaging, single lab","pmids":["42199928"],"is_preprint":false}],"current_model":"CAMSAP3 is a microtubule minus-end binding protein that stabilizes noncentrosomal microtubules by decorating their minus ends (via its CKK domain and MBD, enhanced by C-terminal α-helix-mediated dimerization), thereby anchoring these microtubules at apical cortices of epithelial cells—tethered there via direct interaction with paracingulin (CGNL1) at tight junctions and with ACF7/MACF1—to establish apico-basal microtubule polarity, suppress centrosomal microtubule dominance, orient organelles (Golgi, endosomes, lysosomes), coordinate with dynein/katanin for centrosomal MT release, form the central microtubule pair in motile cilia and kinocilia, maintain nonacetylated microtubules in neuronal axons (suppressing αTAT1-mediated acetylation to preserve single-axon polarity), and scaffold signaling complexes including ERK (via vimentin), Akt, mTORC1, and HMGB1 that control epithelial integrity, mechanosensing, and cell survival."},"narrative":{"mechanistic_narrative":"CAMSAP3 is a microtubule minus-end binding protein that organizes noncentrosomal microtubule arrays, working together with CAMSAP2 to decorate and stabilize minus ends while suppressing centrosomal microtubule-organizing activity [PMID:23169647]. It binds minus ends through its CKK domain, and C-terminal α-helix-mediated dimerization both enhances CKK affinity and enables dynamic regulation of the minus end [PMID:39479887]. By accumulating at and coordinating with dynein and katanin, it drives microtubule release from the centrosome to generate the noncentrosomal pool [PMID:28386021]. In epithelial cells CAMSAP3 concentrates at apical cortices to tether longitudinal microtubule minus ends and establish apico-basal polarity, a localization that requires its CC1 domain and recruitment to tight junctions via direct coiled-coil interaction with paracingulin (CGNL1) [PMID:26715742, PMID:37013686]; apical anchoring and crosstalk with the actin cytoskeleton additionally depend on the spectraplakin ACF7/MACF1 [PMID:27802168, PMID:27693509]. Through this microtubule scaffold CAMSAP3 positions organelles including the Golgi (via CG-NAP) and the endosomal-lysosomal system [PMID:28089391, PMID:41381443]. In neurons it preferentially decorates less-acetylated axonal microtubules and antagonizes αTAT1-mediated tubulin acetylation to enforce single-axon polarity [PMID:30190432]. In multiciliated cells it localizes to the basal body/basal plate region and is required for assembly of the central microtubule pair and synchronized ciliary beating [PMID:32482850, PMID:34319756, PMID:33468623], and it supports transzonal projection architecture and follicle maturation in the ovary [PMID:42199928]. CAMSAP3 also constrains tumor-relevant signaling, suppressing acetylation-driven Akt activity and EMT, sustaining ERK signaling via a vimentin scaffold, and modulating HIF-1α, mTORC1, YAP/PIEZO1, and HMGB1-dependent outputs [PMID:30282632, PMID:33462112, PMID:33712686, PMID:34724356, PMID:37019300, PMID:38598971].","teleology":[{"year":2012,"claim":"Established that CAMSAP3, together with CAMSAP2, defines and stabilizes noncentrosomal microtubules by clustering at their minus ends while suppressing centrosomal microtubule growth, answering how epithelial cells build microtubule arrays not nucleated at the centrosome.","evidence":"siRNA co-depletion with live imaging and immunofluorescence in epithelial cells","pmids":["23169647"],"confidence":"High","gaps":["Did not resolve the molecular basis of minus-end recognition","Did not distinguish CAMSAP2- vs CAMSAP3-specific roles"]},{"year":2015,"claim":"Showed CAMSAP3 tethers microtubule minus ends to the apical cortex via its CC1 domain to orient arrays along the apico-basal axis, connecting minus-end binding to epithelial architecture and organelle positioning.","evidence":"Mutation/knockdown, domain deletion, and forced mislocalization in mouse intestine and Caco-2 cells","pmids":["26715742"],"confidence":"High","gaps":["Did not identify the apical anchor that captures CAMSAP3","Mechanism of CC1-dependent localization unresolved"]},{"year":2016,"claim":"Identified ACF7/MACF1 as a CAMSAP3 partner that anchors minus-end-decorated microtubules to the apical cortex and to F-actin, explaining how the noncentrosomal array is physically coupled to the actin cytoskeleton and migration machinery.","evidence":"Co-IP/pulldown, CAMSAP3 vs CAMSAP2 knockout comparison, live imaging, and migration/focal adhesion assays in two labs","pmids":["27802168","27693509"],"confidence":"High","gaps":["Structural basis of the CAMSAP3-ACF7 interaction not defined","How retrograde actin flow is transmitted to minus ends unresolved"]},{"year":2016,"claim":"Linked CAMSAP3 to Golgi organization and identified the CKK-binding CDH23-C interaction, showing CAMSAP3 microtubule output positions the Golgi and that a disease-associated CDH23 mutation weakens the interaction.","evidence":"Co-IP and knockdown for CG-NAP; in vitro binding, bundling assays and mutagenesis for CDH23-C","pmids":["28089391","27349180"],"confidence":"Medium","gaps":["CG-NAP findings from a single lab with two methods","Functional consequence of CDH23-C regulation of bundling in vivo not established"]},{"year":2017,"claim":"Defined the source of noncentrosomal microtubules by showing CAMSAP3 acts with dynein and katanin to release microtubules from the centrosome, establishing its role upstream of minus-end stabilization.","evidence":"siRNA knockdown, co-depletion epistasis with dynein/katanin, and live imaging in epithelial cells","pmids":["28386021"],"confidence":"Medium","gaps":["Single lab","Order of severing vs CAMSAP3 capping at the centrosome not fully resolved"]},{"year":2018,"claim":"Revealed a neuronal function in which CAMSAP3 maintains a nonacetylated axonal microtubule pool by antagonizing αTAT1, placing it upstream of tubulin acetylation in single-axon polarity, and connected the same acetylation axis to Akt-driven EMT in cancer.","evidence":"Mouse mutation, knockdown/overexpression with αTAT1 epistasis in neurons; deletion with EMT/Akt readouts in lung carcinoma","pmids":["30190432","30282632"],"confidence":"High","gaps":["Mechanism by which CAMSAP3 inhibits αTAT1 not defined","Cancer Akt/EMT findings from a single lab"]},{"year":2020,"claim":"Extended CAMSAP3 function to motile cilia, showing it localizes to basal bodies/basal plate and is required for central microtubule pair assembly and synchronized beating, and to basal body orientation in oviduct MCCs.","evidence":"Mouse hypomorph and Xenopus morpholino knockdown with EM and ciliary beat analysis; mutant mice with super-resolution imaging","pmids":["32482850","34319756","33468623"],"confidence":"High","gaps":["How a minus-end protein templates the central pair mechanistically unclear","Relationship between CAMSAP3 cilia role and PCP signaling only placed parallel/downstream"]},{"year":2021,"claim":"Connected CAMSAP3-dependent microtubule organization to downstream signaling in tissue morphogenesis, linking it to mTORC1/lysosome positioning in ependymal cells and to YAP/PIEZO1 mechanosensing in renal tubules.","evidence":"Camsap3 mutant mice with lysosome/mTORC1 readouts and with YAP/PIEZO1 assays and histology","pmids":["33462112","33712686"],"confidence":"Medium","gaps":["Each pathway shown by a single lab","Direct vs indirect coupling of microtubule arrays to these signaling nodes not dissected"]},{"year":2023,"claim":"Identified paracingulin (CGNL1) as the direct tight-junction receptor that recruits CAMSAP3 to junctions, providing the long-sought apical anchor and confirming junction-tethered minus-end capping by super-resolution imaging.","evidence":"GST pulldown, CGNL1/PLEKHA7 knockout, ultrastructure expansion microscopy, 3D cyst and in vivo intestine","pmids":["37013686"],"confidence":"High","gaps":["How CGNL1-ZO-1 pool coordinates with ACF7 anchoring unresolved","Structure of the CGNL1-CAMSAP3 coiled-coil interface not defined"]},{"year":2024,"claim":"Provided the biochemical basis of minus-end engagement by showing C-terminal α-helix dimerization enhances CKK-domain affinity and enables dynamic minus-end regulation, and extended the cancer scaffolding role to vimentin/ERK, nucleolin/HIF-1α, HMGB1, and EGFR trafficking.","evidence":"In vitro dimerization/MT binding assays with mutagenesis; CRISPR KO with IP-MS, RNA-IP, trafficking and drug-resistance assays in NSCLC","pmids":["39479887","34724356","37019300","38598971","41381443"],"confidence":"Medium","gaps":["Signaling scaffold findings each from single labs","Whether scaffolding is microtubule-dependent or a distinct moonlighting function unresolved"]},{"year":2025,"claim":"Demonstrated an organismal reproductive requirement, showing CAMSAP3 microtubules support granulosa cell transzonal projections and follicle maturation, with knockout mice infertile.","evidence":"Camsap3 knockout mice with super-resolution microscopy and fertility assays","pmids":["42199928"],"confidence":"Medium","gaps":["Single lab","Mechanism linking minus-end organization to TZP stability not defined"]},{"year":null,"claim":"How CAMSAP3's biochemical minus-end activity is integrated across its diverse anchoring partners and signaling scaffolds to produce tissue-specific outcomes remains unresolved.","evidence":"No single study reconciles the minus-end mechanism with the array of context-specific partners and pathways","pmids":[],"confidence":"Medium","gaps":["No unified structural model of CAMSAP3 with its anchors","Whether signaling scaffold roles require microtubule binding is untested across contexts"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,17]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[6,9,10]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[9,10,11]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,2,15]}],"pathway":[{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[9,10,11]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1,7,20]}],"complexes":[],"partners":["CAMSAP2","MACF1","CGNL1","CDH23","AKAP9","VIM","NCL","HMGB1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9P1Y5","full_name":"Calmodulin-regulated spectrin-associated protein 3","aliases":["Protein Nezha"],"length_aa":1249,"mass_kda":134.8,"function":"Key microtubule-organizing protein that specifically binds the minus-end of non-centrosomal microtubules and regulates their dynamics and organization (PubMed:19041755, PubMed:23169647). Specifically recognizes growing microtubule minus-ends and autonomously decorates and stabilizes microtubule lattice formed by microtubule minus-end polymerization (PubMed:24486153). Acts on free microtubule minus-ends that are not capped by microtubule-nucleating proteins or other factors and protects microtubule minus-ends from depolymerization (PubMed:24486153). In addition, it also reduces the velocity of microtubule polymerization (PubMed:24486153). Required for the biogenesis and the maintenance of zonula adherens by anchoring the minus-end of microtubules to zonula adherens and by recruiting the kinesin KIFC3 to those junctional sites (PubMed:19041755). Required for orienting the apical-to-basal polarity of microtubules in epithelial cells: acts by tethering non-centrosomal microtubules to the apical cortex, leading to their longitudinal orientation (PubMed:26715742, PubMed:27802168). Plays a key role in early embryos, which lack centrosomes: accumulates at the microtubule bridges that connect pairs of cells and enables the formation of a non-centrosomal microtubule-organizing center that directs intracellular transport in the early embryo (By similarity). Couples non-centrosomal microtubules with actin: interaction with MACF1 at the minus ends of non-centrosomal microtubules, tethers the microtubules to actin filaments, regulating focal adhesion size and cell migration (PubMed:27693509). Plays a key role in the generation of non-centrosomal microtubules by accumulating in the pericentrosomal region and cooperating with KATNA1 to release non-centrosomal microtubules from the centrosome (PubMed:28386021). Through the microtubule cytoskeleton, also regulates the organization of cellular organelles including the Golgi and the early endosomes (PubMed:28089391). Through interaction with AKAP9, involved in translocation of Golgi vesicles in epithelial cells, where microtubules are mainly non-centrosomal (PubMed:28089391). Plays an important role in motile cilia function by facilitatating proper orientation of basal bodies and formation of central microtubule pairs in motile cilia (By similarity)","subcellular_location":"Cytoplasm, cytoskeleton; Cell junction, adherens junction; Cytoplasm; Cytoplasm, cytoskeleton, cilium axoneme; Cytoplasm, cytoskeleton, cilium basal body","url":"https://www.uniprot.org/uniprotkb/Q9P1Y5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CAMSAP3","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"ARL8B","stoichiometry":0.2},{"gene":"MIS12","stoichiometry":0.2},{"gene":"TRAPPC11","stoichiometry":0.2},{"gene":"TRAPPC2","stoichiometry":0.2},{"gene":"TUBB4B","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/CAMSAP3","total_profiled":1310},"omim":[{"mim_id":"617078","title":"DEATH EFFECTOR DOMAIN-CONTAINING PROTEIN 2; DEDD2","url":"https://www.omim.org/entry/617078"},{"mim_id":"612685","title":"CALMODULIN-REGULATED SPECTRIN-ASSOCIATED PROTEIN 3; CAMSAP3","url":"https://www.omim.org/entry/612685"},{"mim_id":"164176","title":"POU DOMAIN, CLASS 2, TRANSCRIPTION FACTOR 2; POU2F2","url":"https://www.omim.org/entry/164176"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"skin 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and CAMSAP2 cooperate in epithelial-specific organization of noncentrosomal microtubules.","date":"2012","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/23169647","citation_count":131,"is_preprint":false},{"pmid":"26715742","id":"PMC_26715742","title":"CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells.","date":"2015","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/26715742","citation_count":117,"is_preprint":false},{"pmid":"27802168","id":"PMC_27802168","title":"Control of apico-basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7.","date":"2016","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/27802168","citation_count":71,"is_preprint":false},{"pmid":"30190432","id":"PMC_30190432","title":"CAMSAP3 maintains neuronal polarity through regulation of microtubule stability.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30190432","citation_count":62,"is_preprint":false},{"pmid":"27693509","id":"PMC_27693509","title":"The CAMSAP3-ACF7 Complex Couples Noncentrosomal Microtubules with Actin Filaments to Coordinate Their Dynamics.","date":"2016","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/27693509","citation_count":51,"is_preprint":false},{"pmid":"32482850","id":"PMC_32482850","title":"CAMSAP3 facilitates basal body polarity and the formation of the central pair of microtubules in motile cilia.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32482850","citation_count":32,"is_preprint":false},{"pmid":"30282632","id":"PMC_30282632","title":"Loss of CAMSAP3 promotes EMT via the modification of microtubule-Akt machinery.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/30282632","citation_count":32,"is_preprint":false},{"pmid":"18035045","id":"PMC_18035045","title":"Nezha, a novel active miniature inverted-repeat transposable element in cyanobacteria.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/18035045","citation_count":27,"is_preprint":false},{"pmid":"33468623","id":"PMC_33468623","title":"Intercellular and intracellular cilia orientation is coordinated by CELSR1 and CAMSAP3 in oviduct multi-ciliated cells.","date":"2021","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/33468623","citation_count":23,"is_preprint":false},{"pmid":"28386021","id":"PMC_28386021","title":"CAMSAP3 accumulates in the pericentrosomal area and accompanies microtubule release from the centrosome via katanin.","date":"2017","source":"Journal of cell 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bao","url":"https://pubmed.ncbi.nlm.nih.gov/28089391","citation_count":12,"is_preprint":false},{"pmid":"34319756","id":"PMC_34319756","title":"Tracheal motile cilia in mice require CAMSAP3 for the formation of central microtubule pair and coordinated beating.","date":"2021","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/34319756","citation_count":12,"is_preprint":false},{"pmid":"33462112","id":"PMC_33462112","title":"CAMSAP3 is required for mTORC1-dependent ependymal cell growth and lateral ventricle shaping in mouse brains.","date":"2021","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/33462112","citation_count":11,"is_preprint":false},{"pmid":"39206305","id":"PMC_39206305","title":"Adaptability and nutritional analysis of a newly isolated Chlorella sp. NeZha in brackish and marine environments with potential bioeconomic impacts.","date":"2024","source":"Frontiers in nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/39206305","citation_count":5,"is_preprint":false},{"pmid":"38190868","id":"PMC_38190868","title":"CAMSAP3, a microtubule orientation regulator, plays a vital role in manifesting differentiation-dependent characteristics in keratinocytes.","date":"2024","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/38190868","citation_count":4,"is_preprint":false},{"pmid":"35783105","id":"PMC_35783105","title":"Vestibular Hair Cells Require CAMSAP3, a Microtubule Minus-End Regulator, for Formation of Normal Kinocilia.","date":"2022","source":"Frontiers in cellular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/35783105","citation_count":4,"is_preprint":false},{"pmid":"38598971","id":"PMC_38598971","title":"CAMSAP3-mediated regulation of HMGB1 acetylation and subcellular localization in lung cancer cells: Implications for cell death modulation.","date":"2024","source":"Biochimica et biophysica acta. General subjects","url":"https://pubmed.ncbi.nlm.nih.gov/38598971","citation_count":3,"is_preprint":false},{"pmid":"39479887","id":"PMC_39479887","title":"CAMSAP3 forms dimers via its α-helix domain that directly stabilize non-centrosomal microtubule minus ends.","date":"2024","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/39479887","citation_count":2,"is_preprint":false},{"pmid":"41381443","id":"PMC_41381443","title":"Cytoskeletal remodeling via CAMSAP3 downregulation drives resistance to osimertinib in NSCLC cells.","date":"2025","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/41381443","citation_count":0,"is_preprint":false},{"pmid":"42199928","id":"PMC_42199928","title":"Camsap3-mediated microtubules maintain transzonal projections essential for soma-germ communication during ovarian follicle maturation in mice.","date":"2026","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/42199928","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13886,"output_tokens":5865,"usd":0.064816,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14430,"output_tokens":4137,"usd":0.087788,"stage2_stop_reason":"end_turn"},"total_usd":0.152604,"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\": 2012,\n      \"finding\": \"CAMSAP3 (Nezha) and CAMSAP2 co-cluster at the minus ends of noncentrosomal microtubules in epithelial cells, stabilizing them. Depletion of both CAMSAPs caused marked reduction of microtubules with polymerizing plus ends and induced centrosomal microtubule growth, demonstrating that CAMSAP2 and CAMSAP3 cooperatively maintain noncentrosomal microtubules and suppress centrosomal microtubule-organizing activity.\",\n      \"method\": \"siRNA depletion, immunofluorescence, live imaging in epithelial cells\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with specific phenotypic readout replicated across two CAMSAP family members, independent validation of minus-end localization, replicated in subsequent studies\",\n      \"pmids\": [\"23169647\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CAMSAP3 accumulates at apical cortices of intestinal and Caco-2 epithelial cells and tethers longitudinal microtubule minus ends to these sites, thereby orienting microtubule arrays along the apico-basal axis with minus ends apical. The CC1 domain of CAMSAP3 is required for apical localization; forced mislocalization of CAMSAP3 disrupts epithelial architecture. Loss of CAMSAP3 causes random microtubule orientation and perturbed Golgi/nucleus positioning.\",\n      \"method\": \"Camsap3 mutation/siRNA knockdown, domain deletion analysis, forced mislocalization, immunofluorescence in mouse intestinal cells and Caco-2 cells\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function combined with domain mapping and forced mislocalization rescue, replicated in two cell systems\",\n      \"pmids\": [\"26715742\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ACF7 (MACF1), a spectraplakin, specifically binds to CAMSAP3 and is required for apical localization of CAMSAP3-decorated microtubule minus ends in intestinal epithelial cells. Knockout of CAMSAP3 (but not CAMSAP2) caused microtubule reorganization into a radial centrosomal array, redistribution of Rab11-positive endosomes from apical surface to pericentrosomal region, and inhibition of actin brush border formation at the apical side.\",\n      \"method\": \"Co-IP/binding assay, CAMSAP3 knockout, CAMSAP2 knockout, immunofluorescence, 3D cyst culture\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding demonstrated, KO with specific phenotypic readouts, two orthogonal KO comparisons, independently confirmed by a second lab (PMID 27693509)\",\n      \"pmids\": [\"27802168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ACF7 interacts with CAMSAP3 at the minus ends of noncentrosomal microtubules and anchors them to actin filaments in Caco-2 epithelial cells. This CAMSAP3-ACF7 complex enables noncentrosomal microtubules to crosstalk with F-actin via retrograde flow, maintaining microtubule length and perpendicular orientation to the cell edge, and regulating focal adhesion size and cell migration.\",\n      \"method\": \"Co-IP, pulldown, live imaging, siRNA knockdown, focal adhesion/migration assays in Caco-2 cells\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP plus functional phenotypes (migration, focal adhesion), two orthogonal methods, consistent with parallel findings from another lab (PMID 27802168)\",\n      \"pmids\": [\"27693509\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CAMSAP3 binds CG-NAP (centrosome and Golgi localized PKN-associated protein) and plays a role in translocation and clustering of Golgi vesicles in epithelial cells. Depletion of either CAMSAP3 or CG-NAP causes Golgi membrane fragmentation. Stathmin-dependent microtubule dynamics, highest at the perinuclear region, also contributes to perinuclear Golgi distribution.\",\n      \"method\": \"Co-IP, siRNA knockdown, immunofluorescence in epithelial cells\",\n      \"journal\": \"Journal of genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP interaction with CG-NAP and loss-of-function phenotype, single lab, two methods\",\n      \"pmids\": [\"28089391\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CDH23-C (C isoform of cadherin-related 23) directly binds the CKK domain of CAMSAP3 via a conserved N-terminal CKK-binding motif (CBM), and this interaction inhibits CAMSAP3-induced microtubule bundle formation. The Usher Syndrome 1D-associated mutation CDH23 R3175H (mouse R55H) reduces CDH23-C interaction with CAMSAP3 both in vivo and in vitro.\",\n      \"method\": \"In vitro binding assay, pulldown, in vitro MT bundling assay, site-directed mutagenesis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with mutagenesis, single lab\",\n      \"pmids\": [\"27349180\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CAMSAP3 accumulates in the pericentrosomal area and accompanies microtubule release from the centrosome. Depletion of CAMSAP3 prevents MT release and causes MT focusing at centrosomes. CAMSAP3 coordinates with dynein and katanin to regulate MT detachment from the centrosome, identifying it as a key molecule for generation of noncentrosomal microtubules.\",\n      \"method\": \"siRNA knockdown, immunofluorescence, live imaging, co-depletion experiments with dynein and katanin in epithelial cells\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with specific subcellular phenotype, epistatic relationship with katanin/dynein established, single lab\",\n      \"pmids\": [\"28386021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CAMSAP3 is enriched in axons of hippocampal neurons where it localizes preferentially along less-acetylated microtubules. CAMSAP3 mutation causes supernumerary axons and increased nocodazole-resistant/acetylated microtubules. CAMSAP3 depletion promotes tubulin acetylation and overexpression inhibits it, indicating CAMSAP3 retains a nonacetylated microtubule pool by interfering with αTAT1 activity. Depletion of αTAT1 abolished CAMSAP3-loss-dependent multiple-axon formation, placing CAMSAP3 upstream of αTAT1 in neuronal polarity maintenance.\",\n      \"method\": \"Camsap3 mouse mutation, siRNA knockdown, overexpression, αTAT1 depletion epistasis, immunofluorescence, nocodazole resistance assay in hippocampal neurons\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis (αTAT1 rescue), multiple loss-of-function approaches, quantitative phenotypic readouts, in vivo mutation\",\n      \"pmids\": [\"30190432\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CAMSAP3 loss in lung carcinoma cells increases tubulin acetylation, which upregulates Akt (protein kinase B) activity and promotes epithelial-to-mesenchymal transition (EMT) at the transcriptional level. CAMSAP3 therefore functions to suppress Akt activity via microtubule regulation, protecting epithelial phenotype.\",\n      \"method\": \"CAMSAP3 deletion, immunoblotting, EMT marker analysis, Akt activity assay in lung carcinoma cell lines\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with mechanistic pathway (acetylation→Akt→EMT), single lab, two orthogonal readouts\",\n      \"pmids\": [\"30282632\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CAMSAP3 localizes to the base of axonemes and at basal bodies in multiciliated cells (MCCs). In Camsap3 knockdown/hypomorph mice and Xenopus morpholino knockdown, the central microtubule pairs are missing from the majority of cilia, basal body polarity is disorganized, and synchronized ciliary beating is defective. This identifies CAMSAP3 as required for central MT pair assembly/stability in motile cilia.\",\n      \"method\": \"Camsap3 hypomorph mouse model, Xenopus morpholino knockdown, immunofluorescence, electron microscopy, ciliary beat analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — two independent model organisms (mouse and Xenopus), electron microscopy structural validation, loss-of-function with defined phenotype, replicated in subsequent studies (PMIDs 34319756, 35783105)\",\n      \"pmids\": [\"32482850\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3 concentrates at multiple sites of the cilium-basal body complex including the axonemal basal plate where the central pair of microtubules (CP) initiates. CAMSAP3 dysfunction results in loss of the CP, partial distortion of the basal plate, and failure of multicilia to undergo synchronized beating in tracheal airway epithelial cells.\",\n      \"method\": \"Camsap3 mutant mouse model, immunofluorescence, electron microscopy, high-speed videomicroscopy of ciliary beating\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with structural (EM) and functional (beat) readouts, consistent with independent findings (PMID 32482850)\",\n      \"pmids\": [\"34319756\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In mouse oviduct MCCs, CAMSAP3 localizes to the base of cilia in a polarized manner and is critical for intracellular basal body (BB) orientation and for assembly of microtubules interconnecting basal bodies. CAMSAP3 mutation disrupts intracellular coordination of BB orientation without affecting planar cell polarity (PCP) factor localization, placing CAMSAP3 function downstream of or parallel to PCP signaling.\",\n      \"method\": \"Camsap3 mutant mice, immunofluorescence, super-resolution microscopy, genetic comparison with CELSR1 mutants\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with structural readout, epistatic placement relative to PCP, single lab\",\n      \"pmids\": [\"33468623\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In ependymal cells, CAMSAP3 concentrates at apical domains to generate MT networks. Camsap3-mutated mice exhibit narrowed lateral ventricles due to failure of ependymal cells to broaden their apical domain. mTORC1 activity, required for ependymal cell growth, is downregulated in mutant cells, and lysosomes (which activate mTORC1) are reduced at apical regions where MT networks are disorganized, linking CAMSAP3-dependent MT organization to mTORC1 signaling and ventricle shaping.\",\n      \"method\": \"Camsap3 mutant mouse, immunofluorescence, lysosome localization assay, mTORC1 activity measurement, apical domain morphometry\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with pathway mechanistic follow-up (mTORC1/lysosome), single lab, two orthogonal mechanistic readouts\",\n      \"pmids\": [\"33462112\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In mouse proximal convoluted tubules, CAMSAP3 mutation perturbs microtubule arrays and activates mechanosensors YAP and PIEZO1, leading to enhanced cell proliferation, cell flattening, and cyst formation. This places CAMSAP3-mediated microtubule networks upstream of mechanosensitive YAP/PIEZO1 signaling in renal tubule morphology maintenance.\",\n      \"method\": \"Camsap3 mutant mice, immunofluorescence, YAP/PIEZO1 activity assays, histology\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with mechanistic pathway placement (YAP/PIEZO1), single lab\",\n      \"pmids\": [\"33712686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3 knockout promotes lung cancer cell senescence-associated phenotypes and G1 cell cycle arrest. Mechanistically, phosphorylated ERK is markedly downregulated in CAMSAP3-deleted cells, suppressing cyclin D1. Proteomic analysis identified vimentin as a scaffold required for CAMSAP3-modulated ERK signaling. Full-length CAMSAP3 abrogates these phenotypes.\",\n      \"method\": \"CAMSAP3 CRISPR knockout, immunoprecipitation/mass spectrometry, Western blotting, xenograft model\",\n      \"journal\": \"Cancer medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO with proteomic identification of scaffold (vimentin), in vivo validation, single lab\",\n      \"pmids\": [\"34724356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Paracingulin (CGNL1), but not PLEKHA7, recruits CAMSAP3 to tight junctions. GST pulldown shows CGNL1 interacts directly with CAMSAP3 via their respective coiled-coil regions. Knockout of CGNL1 causes loss of junctional CAMSAP3, disorganized cytoplasmic microtubules, irregular nuclei alignment, altered cyst morphogenesis, and disrupted planar apical microtubules. Ultrastructure expansion microscopy shows CAMSAP3-capped microtubules are tethered to junctions by the ZO-1-associated pool of CGNL1.\",\n      \"method\": \"GST pulldown, CGNL1/PLEKHA7 knockout, ultrastructure expansion microscopy, immunofluorescence, 3D cyst culture, in vivo mouse intestinal epithelium\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct biochemical interaction (GST pulldown) combined with KO phenotype and super-resolution structural imaging, two orthogonal methods, in vitro and in vivo validation\",\n      \"pmids\": [\"37013686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CAMSAP3 interacts with nucleolin (NCL) to regulate HIF-1α mRNA stabilization. In CAMSAP3-knockout NSCLC cells, HIF-1α expression is upregulated along with VEGFA and MMPs 2 and 9, increasing invasion and angiogenesis. These effects were attenuated by reintroduction of wild-type CAMSAP3.\",\n      \"method\": \"CAMSAP3 CRISPR knockout, immunoprecipitation, mass spectrometry, RNA immunoprecipitation, qRT-PCR, in vivo xenograft\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interaction with NCL identified by MS, RNA-IP for mRNA stabilization, rescue experiment, single lab\",\n      \"pmids\": [\"37019300\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 forms dimers through its C-terminal α-helix domain. This dimerization enhances the microtubule-binding affinity of the CKK domain and enables the CKK domain to regulate microtubule dynamics. The combined action of the microtubule-binding domain (MBD) and the C-terminal α-helix enables CAMSAP3 to decorate minus ends and achieve dynamic regulation of microtubule minus ends.\",\n      \"method\": \"Biochemical dimerization assays, domain deletion/mutagenesis, in vitro microtubule binding and dynamics assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with domain mutagenesis establishing dimerization mechanism and CKK domain enhancement, single lab but multiple orthogonal biochemical methods\",\n      \"pmids\": [\"39479887\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 interacts with HMGB1, particularly with its acetylated form. In the presence of the HDAC inhibitor trichostatin A (TSA), CAMSAP3-HMGB1 complex formation is elevated, facilitating HMGB1 cytoplasmic translocation and secretion, thereby inducing autophagic cell death. CAMSAP3 knockout abolishes TSA-mediated autophagic cell death in lung cancer cells.\",\n      \"method\": \"CRISPR-Cas9 knockout, immunoprecipitation, proteomics, immunofluorescence, autophagic detection assays\",\n      \"journal\": \"Biochimica et biophysica acta. General subjects\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interaction with acetylated HMGB1 and KO functional phenotype, two orthogonal methods, single lab\",\n      \"pmids\": [\"38598971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 is essential for proper localization of the microtubule-dependent endosomal-lysosomal system. CAMSAP3 depletion causes EGFR translocation to the perinuclear MTOC, blocking plasma membrane recycling and promoting lysosomal EGFR degradation, thereby reducing EGFR signaling and increasing osimertinib resistance in NSCLC.\",\n      \"method\": \"CAMSAP3 siRNA knockdown and overexpression, immunofluorescence, in vitro and xenograft drug sensitivity assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function and gain-of-function with defined subcellular trafficking phenotype, in vivo xenograft validation, single lab\",\n      \"pmids\": [\"41381443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Camsap3-mediated microtubules are present in transzonal projections (TZPs) of granulosa cells extending to oocytes in ovarian follicles. Camsap3 knockout mice are infertile and lack ovulation; their follicles show reduced TZP numbers, disorganized TZP microtubules, and decreased granulosa cell-oocyte contact, identifying CAMSAP3 as required for TZP architecture and follicle maturation.\",\n      \"method\": \"Camsap3 knockout mice, super-resolution microscopy, follicle counting, fertility assays\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with specific structural and functional phenotypes, super-resolution imaging, single lab\",\n      \"pmids\": [\"42199928\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CAMSAP3 is a microtubule minus-end binding protein that stabilizes noncentrosomal microtubules by decorating their minus ends (via its CKK domain and MBD, enhanced by C-terminal α-helix-mediated dimerization), thereby anchoring these microtubules at apical cortices of epithelial cells—tethered there via direct interaction with paracingulin (CGNL1) at tight junctions and with ACF7/MACF1—to establish apico-basal microtubule polarity, suppress centrosomal microtubule dominance, orient organelles (Golgi, endosomes, lysosomes), coordinate with dynein/katanin for centrosomal MT release, form the central microtubule pair in motile cilia and kinocilia, maintain nonacetylated microtubules in neuronal axons (suppressing αTAT1-mediated acetylation to preserve single-axon polarity), and scaffold signaling complexes including ERK (via vimentin), Akt, mTORC1, and HMGB1 that control epithelial integrity, mechanosensing, and cell survival.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CAMSAP3 is a microtubule minus-end binding protein that organizes noncentrosomal microtubule arrays, working together with CAMSAP2 to decorate and stabilize minus ends while suppressing centrosomal microtubule-organizing activity [#0]. It binds minus ends through its CKK domain, and C-terminal \\u03b1-helix-mediated dimerization both enhances CKK affinity and enables dynamic regulation of the minus end [#17]. By accumulating at and coordinating with dynein and katanin, it drives microtubule release from the centrosome to generate the noncentrosomal pool [#6]. In epithelial cells CAMSAP3 concentrates at apical cortices to tether longitudinal microtubule minus ends and establish apico-basal polarity, a localization that requires its CC1 domain and recruitment to tight junctions via direct coiled-coil interaction with paracingulin (CGNL1) [#1, #15]; apical anchoring and crosstalk with the actin cytoskeleton additionally depend on the spectraplakin ACF7/MACF1 [#2, #3]. Through this microtubule scaffold CAMSAP3 positions organelles including the Golgi (via CG-NAP) and the endosomal-lysosomal system [#4, #19]. In neurons it preferentially decorates less-acetylated axonal microtubules and antagonizes \\u03b1TAT1-mediated tubulin acetylation to enforce single-axon polarity [#7]. In multiciliated cells it localizes to the basal body/basal plate region and is required for assembly of the central microtubule pair and synchronized ciliary beating [#9, #10, #11], and it supports transzonal projection architecture and follicle maturation in the ovary [#20]. CAMSAP3 also constrains tumor-relevant signaling, suppressing acetylation-driven Akt activity and EMT, sustaining ERK signaling via a vimentin scaffold, and modulating HIF-1\\u03b1, mTORC1, YAP/PIEZO1, and HMGB1-dependent outputs [#8, #12, #13, #14, #16, #18].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established that CAMSAP3, together with CAMSAP2, defines and stabilizes noncentrosomal microtubules by clustering at their minus ends while suppressing centrosomal microtubule growth, answering how epithelial cells build microtubule arrays not nucleated at the centrosome.\",\n      \"evidence\": \"siRNA co-depletion with live imaging and immunofluorescence in epithelial cells\",\n      \"pmids\": [\"23169647\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the molecular basis of minus-end recognition\", \"Did not distinguish CAMSAP2- vs CAMSAP3-specific roles\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showed CAMSAP3 tethers microtubule minus ends to the apical cortex via its CC1 domain to orient arrays along the apico-basal axis, connecting minus-end binding to epithelial architecture and organelle positioning.\",\n      \"evidence\": \"Mutation/knockdown, domain deletion, and forced mislocalization in mouse intestine and Caco-2 cells\",\n      \"pmids\": [\"26715742\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the apical anchor that captures CAMSAP3\", \"Mechanism of CC1-dependent localization unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identified ACF7/MACF1 as a CAMSAP3 partner that anchors minus-end-decorated microtubules to the apical cortex and to F-actin, explaining how the noncentrosomal array is physically coupled to the actin cytoskeleton and migration machinery.\",\n      \"evidence\": \"Co-IP/pulldown, CAMSAP3 vs CAMSAP2 knockout comparison, live imaging, and migration/focal adhesion assays in two labs\",\n      \"pmids\": [\"27802168\", \"27693509\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the CAMSAP3-ACF7 interaction not defined\", \"How retrograde actin flow is transmitted to minus ends unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Linked CAMSAP3 to Golgi organization and identified the CKK-binding CDH23-C interaction, showing CAMSAP3 microtubule output positions the Golgi and that a disease-associated CDH23 mutation weakens the interaction.\",\n      \"evidence\": \"Co-IP and knockdown for CG-NAP; in vitro binding, bundling assays and mutagenesis for CDH23-C\",\n      \"pmids\": [\"28089391\", \"27349180\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CG-NAP findings from a single lab with two methods\", \"Functional consequence of CDH23-C regulation of bundling in vivo not established\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined the source of noncentrosomal microtubules by showing CAMSAP3 acts with dynein and katanin to release microtubules from the centrosome, establishing its role upstream of minus-end stabilization.\",\n      \"evidence\": \"siRNA knockdown, co-depletion epistasis with dynein/katanin, and live imaging in epithelial cells\",\n      \"pmids\": [\"28386021\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Order of severing vs CAMSAP3 capping at the centrosome not fully resolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Revealed a neuronal function in which CAMSAP3 maintains a nonacetylated axonal microtubule pool by antagonizing \\u03b1TAT1, placing it upstream of tubulin acetylation in single-axon polarity, and connected the same acetylation axis to Akt-driven EMT in cancer.\",\n      \"evidence\": \"Mouse mutation, knockdown/overexpression with \\u03b1TAT1 epistasis in neurons; deletion with EMT/Akt readouts in lung carcinoma\",\n      \"pmids\": [\"30190432\", \"30282632\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which CAMSAP3 inhibits \\u03b1TAT1 not defined\", \"Cancer Akt/EMT findings from a single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Extended CAMSAP3 function to motile cilia, showing it localizes to basal bodies/basal plate and is required for central microtubule pair assembly and synchronized beating, and to basal body orientation in oviduct MCCs.\",\n      \"evidence\": \"Mouse hypomorph and Xenopus morpholino knockdown with EM and ciliary beat analysis; mutant mice with super-resolution imaging\",\n      \"pmids\": [\"32482850\", \"34319756\", \"33468623\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a minus-end protein templates the central pair mechanistically unclear\", \"Relationship between CAMSAP3 cilia role and PCP signaling only placed parallel/downstream\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected CAMSAP3-dependent microtubule organization to downstream signaling in tissue morphogenesis, linking it to mTORC1/lysosome positioning in ependymal cells and to YAP/PIEZO1 mechanosensing in renal tubules.\",\n      \"evidence\": \"Camsap3 mutant mice with lysosome/mTORC1 readouts and with YAP/PIEZO1 assays and histology\",\n      \"pmids\": [\"33462112\", \"33712686\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Each pathway shown by a single lab\", \"Direct vs indirect coupling of microtubule arrays to these signaling nodes not dissected\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified paracingulin (CGNL1) as the direct tight-junction receptor that recruits CAMSAP3 to junctions, providing the long-sought apical anchor and confirming junction-tethered minus-end capping by super-resolution imaging.\",\n      \"evidence\": \"GST pulldown, CGNL1/PLEKHA7 knockout, ultrastructure expansion microscopy, 3D cyst and in vivo intestine\",\n      \"pmids\": [\"37013686\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CGNL1-ZO-1 pool coordinates with ACF7 anchoring unresolved\", \"Structure of the CGNL1-CAMSAP3 coiled-coil interface not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided the biochemical basis of minus-end engagement by showing C-terminal \\u03b1-helix dimerization enhances CKK-domain affinity and enables dynamic minus-end regulation, and extended the cancer scaffolding role to vimentin/ERK, nucleolin/HIF-1\\u03b1, HMGB1, and EGFR trafficking.\",\n      \"evidence\": \"In vitro dimerization/MT binding assays with mutagenesis; CRISPR KO with IP-MS, RNA-IP, trafficking and drug-resistance assays in NSCLC\",\n      \"pmids\": [\"39479887\", \"34724356\", \"37019300\", \"38598971\", \"41381443\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Signaling scaffold findings each from single labs\", \"Whether scaffolding is microtubule-dependent or a distinct moonlighting function unresolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated an organismal reproductive requirement, showing CAMSAP3 microtubules support granulosa cell transzonal projections and follicle maturation, with knockout mice infertile.\",\n      \"evidence\": \"Camsap3 knockout mice with super-resolution microscopy and fertility assays\",\n      \"pmids\": [\"42199928\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Mechanism linking minus-end organization to TZP stability not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CAMSAP3's biochemical minus-end activity is integrated across its diverse anchoring partners and signaling scaffolds to produce tissue-specific outcomes remains unresolved.\",\n      \"evidence\": \"No single study reconciles the minus-end mechanism with the array of context-specific partners and pathways\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified structural model of CAMSAP3 with its anchors\", \"Whether signaling scaffold roles require microtubule binding is untested across contexts\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 17]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [6, 9, 10]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [9, 10, 11]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 2, 15]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [9, 10, 11]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1, 7, 20]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"CAMSAP2\", \"MACF1\", \"CGNL1\", \"CDH23\", \"AKAP9\", \"VIM\", \"NCL\", \"HMGB1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}