{"gene":"CAMSAP3","run_date":"2026-04-28T17:28:52","timeline":{"discoveries":[{"year":2012,"finding":"CAMSAP3 (Nezha) and CAMSAP2 co-cluster at the minus ends of noncentrosomal microtubules in epithelial cells, stabilizing them and suppressing centrosomal microtubule organization; depletion of both CAMSAPs caused marked reduction of microtubules with polymerizing plus ends and perturbed early endosome and Golgi distribution.","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 — clean KD with defined cellular phenotypes, replicated across two CAMSAP family members, highly cited foundational study","pmids":["23169647"],"is_preprint":false},{"year":2015,"finding":"CAMSAP3 accumulates at apical cortices of polarized intestinal epithelial cells and tethers longitudinal microtubule minus ends there, orienting microtubules along the apical-to-basal axis; its CC1 domain is required for apical localization, and forced mislocalization disrupts epithelial architecture.","method":"Camsap3 mutation/depletion in mouse intestinal cells and Caco-2 cells, domain mutagenesis, immunofluorescence, forced mislocalization experiments","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with defined phenotype plus domain mutagenesis, replicated in two cell systems","pmids":["26715742"],"is_preprint":false},{"year":2016,"finding":"ACF7 (MACF1), a spectraplakin family cytoskeletal crosslinking protein, specifically binds CAMSAP3 and is required for apical localization of CAMSAP3-decorated microtubule minus ends in intestinal epithelial cells; CAMSAP3 knockout (but not CAMSAP2) caused microtubule reorganization into a centrosomal radial array and redistribution of Rab11-positive endosomes away from the apical surface.","method":"Reciprocal Co-IP, knockout cell lines, immunofluorescence, 3D cyst culture","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP plus KO with multiple defined phenotypes","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; this CAMSAP3-ACF7 complex couples microtubule and actin dynamics to regulate retrograde flow, focal adhesion size, and cell migration in Caco-2 epithelial cells.","method":"Co-IP, pulldown, live imaging, siRNA depletion, focal adhesion and migration assays","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 — Co-IP plus loss-of-function with multiple orthogonal phenotypic readouts","pmids":["27693509"],"is_preprint":false},{"year":2017,"finding":"CAMSAP3 accumulates in the pericentrosomal area and cooperates with dynein and katanin to mediate microtubule detachment from the centrosome; depletion of CAMSAP3 prevents microtubule release and causes focusing of microtubules at centrosomes.","method":"siRNA depletion, immunofluorescence, live imaging, Co-IP with dynein and katanin","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function with defined phenotype and Co-IP with molecular partners, single lab","pmids":["28386021"],"is_preprint":false},{"year":2016,"finding":"CAMSAP3 interacts with CG-NAP and regulates Golgi vesicle translocation in epithelial cells; depletion of either CAMSAP3 or CG-NAP causes Golgi membrane fragmentation, and stathmin-dependent microtubule dynamics graded along the radial axis contributes to perinuclear Golgi assembly.","method":"Co-IP, siRNA depletion, immunofluorescence, Golgi morphology assays","journal":"Journal of genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus loss-of-function with defined Golgi phenotype, single lab","pmids":["28089391"],"is_preprint":false},{"year":2018,"finding":"CAMSAP3 is enriched in axons of hippocampal neurons and preferentially localizes to less-acetylated microtubules; CAMSAP3 mutation causes supernumerary axons and increased tubulin acetylation; CAMSAP3 depletion promotes αTAT1-dependent tubulin acetylation, and αTAT1 depletion abolishes multi-axon formation caused by CAMSAP3 loss, placing CAMSAP3 upstream of αTAT1 in axonal polarity maintenance.","method":"Genetic epistasis (double KD), mouse mutants, immunofluorescence, nocodazole resistance assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — epistasis experiment plus loss-of-function in primary neurons with multiple defined phenotypes","pmids":["30190432"],"is_preprint":false},{"year":2018,"finding":"Loss of CAMSAP3 in lung carcinoma cells increases tubulin acetylation, which upregulates Akt activity and promotes EMT at the transcriptional level; CAMSAP3 thus protects epithelial phenotype by suppressing Akt activity via microtubule regulation.","method":"CAMSAP3 deletion cell lines, Akt activity assays, tubulin acetylation immunoblot, EMT marker analysis","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — KO with mechanistic pathway placement, single lab","pmids":["30282632"],"is_preprint":false},{"year":2020,"finding":"CAMSAP3 localizes to the base of axonemes and at basal bodies in multiciliated cells; its loss results in absence of the central microtubule pair in most cilia and disorganized basal body polarity, causing defective synchronized ciliary beating and PCD-like phenotypes; confirmed in Xenopus by morpholino knockdown.","method":"Hypomorphic knockdown mouse model, Xenopus morpholino, electron microscopy, immunofluorescence, ciliary beat analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function in two model organisms with ultrastructural and functional validation","pmids":["32482850"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 concentrates at the axonemal basal plate (transition zone) where the central microtubule pair initiates; CAMSAP3 dysfunction causes loss of the central pair and partial distortion of the basal plate, as well as failure of coordinated multicilia beating in tracheal airway epithelial cells.","method":"Mouse mutants, super-resolution immunofluorescence, electron microscopy, ciliary beat analysis","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 — ultrastructural localization plus loss-of-function with defined structural and functional phenotypes","pmids":["34319756"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 localizes to the base of cilia in a polarized manner in oviduct multiciliated cells; its mutation disrupts intracellular coordination of basal body orientation and the assembly of microtubules interconnecting basal bodies, without affecting planar cell polarity factor localization, indicating a distinct mechanism from CELSR1.","method":"Mouse mutants, immunofluorescence, super-resolution microscopy, basal body orientation quantification","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis/pathway separation plus loss-of-function with structural phenotype","pmids":["33468623"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 concentrates at apical domains of differentiating ependymal cells and generates apical MT networks that support lysosomal positioning; CAMSAP3 mutation downregulates mTORC1 activity, reduces apical lysosome distribution, and impairs ependymal cell apical domain expansion, leading to abnormal lateral ventricle narrowing.","method":"Camsap3 mutant mice, mTORC1 activity assays, lysosome immunofluorescence, ventricle morphometry","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function in mouse with mechanistic pathway placement, single lab","pmids":["33462112"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3-mediated microtubule networks are required to maintain mechanical properties of proximal convoluted tubule (PCT) cells; CAMSAP3 mutation activates mechanosensors YAP and PIEZO1, induces cell flattening and proliferation, and causes PCT cyst formation.","method":"Camsap3 mutant mice, YAP/PIEZO1 immunostaining, electron microscopy, proliferation assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — mouse loss-of-function with mechanosensor pathway placement, single lab","pmids":["33712686"],"is_preprint":false},{"year":2021,"finding":"CAMSAP3 knockout promotes lung cancer cell senescence-associated phenotypes and G1 arrest via downregulation of phospho-ERK and cyclin D1; vimentin acts as a scaffold for the CAMSAP3-ERK signaling complex, identified by immunoprecipitation/mass spectrometry.","method":"CRISPR-Cas9 KO, IP/mass spectrometry, immunoblotting, flow cytometry, xenograft model","journal":"Cancer medicine","confidence":"Medium","confidence_rationale":"Tier 2 — KO with mechanistic pathway placement and proteomic partner identification, single lab","pmids":["34724356"],"is_preprint":false},{"year":2016,"finding":"The C isoform of CDH23 directly binds the CKK domain of CAMSAP3 via a conserved N-terminal CKK-binding motif (CBM) and inhibits CAMSAP3-induced microtubule bundle formation; the Usher Syndrome 1D mutation CDH23 R3175H (mouse R55H) reduces this interaction both in vitro and in vivo.","method":"Co-IP, pulldown, in vitro binding assays, mutagenesis, in vivo mouse models","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1/2 — direct binding with mutagenesis and in vivo validation","pmids":["27349180"],"is_preprint":false},{"year":2023,"finding":"Paracingulin (CGNL1), but not PLEKHA7, interacts with CAMSAP3 via their coiled-coil regions (GST pulldown) and recruits CAMSAP3 to tight junctions via the ZO-1-associated pool; CGNL1 knockout causes loss of junctional CAMSAP3, disorganized cytoplasmic microtubules, and disrupted epithelial architecture.","method":"GST pulldown, KO cell lines, ultrastructure expansion microscopy, in vivo mouse intestinal epithelium","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1/2 — GST pulldown defines interaction domain plus KO with multiple orthogonal phenotypes in vitro and in vivo","pmids":["37013686"],"is_preprint":false},{"year":2023,"finding":"CAMSAP3 interacts with nucleolin (NCL) to regulate HIF-1α mRNA stabilization; CAMSAP3 knockout increases HIF-1α expression and downstream VEGFA and MMP2/9, promoting invasion and angiogenesis; identified by proteomic analysis and RNA immunoprecipitation.","method":"CRISPR-Cas9 KO, IP/mass spectrometry, RNA immunoprecipitation, in vitro invasion/angiogenesis assays, in vivo xenograft","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 — proteomic interaction plus RNA-IP with loss-of-function, single lab","pmids":["37019300"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 forms dimers through its C-terminal α-helix domain; dimerization enhances the microtubule-binding affinity of the CKK domain and enables the CKK domain to regulate minus-end dynamics; the combined action of the microtubule-binding domain (MBD) and C-terminal α-helix mediates minus-end decoration and dynamic regulation.","method":"Biochemical dimerization assays, in vitro microtubule-binding assays, domain mutagenesis, live cell imaging","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with domain mutagenesis defining molecular mechanism","pmids":["39479887"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 interacts with acetylated HMGB1 in the cytoplasm; TSA-induced HMGB1 acetylation drives HMGB1 cytoplasmic translocation and secretion promoting autophagic cell death, a process that requires CAMSAP3; CAMSAP3 knockout abolishes this TSA-mediated autophagic cell death.","method":"CRISPR-Cas9 KO, proteomic analysis, immunoprecipitation, immunofluorescence, autophagic detection assays","journal":"Biochimica et biophysica acta. General subjects","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus KO with defined functional outcome, single lab","pmids":["38598971"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 depletion in NSCLC cells causes centrosomal microtubule clustering, impairs the endosomal-lysosomal system, and drives EGFR translocation to the perinuclear MTOC, blocking plasma membrane recycling and promoting lysosomal EGFR degradation, thereby conferring osimertinib resistance; CAMSAP3 overexpression in resistant cells restores microtubule organization and drug sensitivity.","method":"siRNA/CRISPR depletion, CAMSAP3 overexpression, EGFR trafficking assays, xenograft model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 — bidirectional genetic manipulation with defined mechanistic pathway, single lab","pmids":["41381443"],"is_preprint":false},{"year":2024,"finding":"CAMSAP3 deficiency in keratinocytes shifts microtubules from non-centrosomal to centrosomal configuration, causes cell cycle exit, delayed cytokinesis, and impairs formation and stability of adherens junctions and tight junctions; re-expression of CAMSAP3 rescues these defects.","method":"Inducible CAMSAP3 KO and re-expression in HaCaT cells, immunofluorescence, flow cytometry","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — bidirectional genetic rescue experiment with multiple phenotypic readouts, single lab","pmids":["38190868"],"is_preprint":false},{"year":2022,"finding":"CAMSAP3 is required for formation and/or maintenance of the central microtubule pair in vestibular hair cell kinocilia; conditional knockout of CAMSAP3 results in shorter kinocilia and more frequent absence of the central MT pair, linking CAMSAP3 to axoneme length and stability.","method":"Conditional KO mouse model, immunofluorescence, transmission electron microscopy, behavioral assays","journal":"Frontiers in cellular neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function with ultrastructural phenotype, single lab","pmids":["35783105"],"is_preprint":false},{"year":2025,"finding":"CAMSAP3-mediated microtubules are required for maintenance of transzonal projections (TZPs) between granulosa cells and oocytes in ovarian follicles; CAMSAP3 KO mice are infertile with fewer late-stage follicles, reduced TZP number, and disorganized microtubules in TZPs; CAMSAP3 also modulates TZP morphology by organizing both microtubules and F-actin.","method":"CAMSAP3 KO mice, super-resolution microscopy, TZP quantification, follicle staging","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — KO with defined ultrastructural and fertility phenotypes; preprint, not yet peer-reviewed","pmids":["bio_10.1101_2025.09.26.678897"],"is_preprint":true}],"current_model":"CAMSAP3 is a microtubule minus-end binding protein that forms dimers via its C-terminal α-helix (enhancing CKK domain binding affinity) and stabilizes noncentrosomal microtubule minus ends through its MBD and CKK domains; it localizes to apical cortices, pericentrosomal regions, tight junctions (via paracingulin/CGNL1), and the axonemal basal plate, where it cooperates with ACF7/MACF1 to anchor minus ends to actin, with dynein and katanin to release microtubules from the centrosome, and with CDH23-C to modulate microtubule bundling, thereby orienting apico-basal microtubule polarity in epithelia, maintaining neuronal polarity by suppressing αTAT1-dependent tubulin acetylation, forming the central microtubule pair in motile cilia and kinocilia, and regulating downstream signaling through Akt, ERK/cyclin D1, and mTORC1 pathways."},"narrative":{"teleology":[{"year":2012,"claim":"Establishing that CAMSAP3 and CAMSAP2 specifically recognize and stabilize noncentrosomal microtubule minus ends answered the fundamental question of how epithelial cells maintain acentrosomal microtubule networks and organize endomembranes.","evidence":"siRNA depletion with immunofluorescence and live imaging in epithelial cells","pmids":["23169647"],"confidence":"High","gaps":["Mechanism of minus-end recognition not defined at the molecular level","Relative contributions of CAMSAP2 vs CAMSAP3 not separated","How microtubule stabilization suppresses centrosomal organization unclear"]},{"year":2015,"claim":"Demonstrating that CAMSAP3 accumulates at apical cortices via its CC1 domain and tethers minus ends there established the molecular basis of apico-basal microtubule orientation in intestinal epithelia.","evidence":"Domain mutagenesis and forced mislocalization in mouse intestinal cells and Caco-2 cells","pmids":["26715742"],"confidence":"High","gaps":["Identity of the apical receptor for the CC1 domain unknown","Whether CC1-mediated localization is conserved across all polarized epithelia not tested"]},{"year":2016,"claim":"Identification of ACF7/MACF1 as a direct CAMSAP3 partner that anchors minus-end-decorated microtubules to actin filaments resolved how the noncentrosomal microtubule network is physically connected to the cortical cytoskeleton and influences focal adhesion dynamics and migration.","evidence":"Reciprocal Co-IP, pulldown, KO cell lines, live imaging, focal adhesion and migration assays in Caco-2 cells and 3D cysts","pmids":["27802168","27693509"],"confidence":"High","gaps":["Structural basis of CAMSAP3–ACF7 interaction not mapped to specific domains","Contribution of other spectraplakins not excluded"]},{"year":2016,"claim":"Discovery that CDH23-C directly binds the CKK domain via a conserved CBM motif and inhibits CAMSAP3-induced microtubule bundling linked CAMSAP3 to stereocilia/inner ear biology and the Usher Syndrome 1D mutation R3175H.","evidence":"Co-IP, in vitro pulldown, mutagenesis, in vivo mouse models","pmids":["27349180"],"confidence":"High","gaps":["Physiological consequence of CDH23–CAMSAP3 disruption on hearing not directly shown","Whether other CKK-binding partners compete with CDH23 unknown"]},{"year":2016,"claim":"Identification of CAMSAP3 interaction with CG-NAP and its role in Golgi vesicle translocation provided the first link between CAMSAP3-organized microtubules and Golgi ribbon integrity.","evidence":"Co-IP, siRNA depletion, Golgi morphology assays in epithelial cells","pmids":["28089391"],"confidence":"Medium","gaps":["Direct binding interface between CAMSAP3 and CG-NAP not mapped","Golgi phenotype not validated in vivo","Whether CAMSAP2 compensates at the Golgi not tested"]},{"year":2017,"claim":"Showing that CAMSAP3 cooperates with dynein and katanin at the pericentrosomal area to release microtubules from centrosomes answered how noncentrosomal microtubules are generated from centrosomal precursors.","evidence":"siRNA depletion, live imaging, Co-IP with dynein and katanin","pmids":["28386021"],"confidence":"Medium","gaps":["Order of recruitment (CAMSAP3 vs dynein vs katanin) not resolved","Reconstitution of microtubule release in vitro not achieved","Single lab finding"]},{"year":2018,"claim":"Genetic epistasis placing CAMSAP3 upstream of αTAT1-dependent tubulin acetylation in maintaining single-axon polarity established a new regulatory axis: CAMSAP3 suppresses tubulin acetylation to restrict axon number.","evidence":"Double knockdown epistasis, mouse Camsap3 mutants, nocodazole resistance assays in hippocampal neurons","pmids":["30190432"],"confidence":"High","gaps":["Molecular mechanism by which CAMSAP3 inhibits αTAT1 access not defined","Whether this axis operates in other neuronal subtypes not tested"]},{"year":2018,"claim":"Linking CAMSAP3 loss to increased tubulin acetylation, Akt activation, and EMT in lung carcinoma extended the tubulin acetylation axis to signaling and cancer biology, showing CAMSAP3 protects epithelial identity.","evidence":"CAMSAP3 deletion cell lines, Akt activity assays, EMT marker analysis","pmids":["30282632"],"confidence":"Medium","gaps":["Direct mechanism connecting tubulin acetylation to Akt activation not delineated","In vivo tumor progression data lacking at this point"]},{"year":2020,"claim":"Demonstrating that CAMSAP3 localizes to axonemal bases and is required for central microtubule pair formation in motile cilia revealed an unexpected structural role, explaining PCD-like phenotypes upon CAMSAP3 loss.","evidence":"Hypomorphic knockdown mouse, Xenopus morpholino, electron microscopy, ciliary beat analysis","pmids":["32482850"],"confidence":"High","gaps":["How CAMSAP3 nucleates or stabilizes the central pair specifically is unknown","Whether CAMSAP3 acts alone or requires cofactors at the basal plate not resolved"]},{"year":2021,"claim":"Refining CAMSAP3 localization to the basal plate (transition zone) and demonstrating that its loss distorts basal plate ultrastructure and disrupts basal body coordination in multiciliated airways and oviduct cells consolidated the ciliary role and separated it mechanistically from planar cell polarity signaling via CELSR1.","evidence":"Mouse mutants, super-resolution microscopy, electron microscopy, basal body orientation quantification in tracheal and oviduct cells","pmids":["34319756","33468623"],"confidence":"High","gaps":["Basal plate binding partners of CAMSAP3 not identified","Whether CAMSAP3 functions at primary (non-motile) cilia not tested"]},{"year":2021,"claim":"Connecting CAMSAP3-organized microtubules to lysosomal positioning, mTORC1 signaling, and ependymal cell apical domain expansion broadened the downstream effector repertoire beyond acetylation-Akt, and showed tissue-level consequences (ventricle narrowing) of CAMSAP3 loss.","evidence":"Camsap3 mutant mice, mTORC1 activity assays, lysosome immunofluorescence, ventricle morphometry","pmids":["33462112"],"confidence":"Medium","gaps":["Whether mTORC1 downregulation is a direct consequence of lysosome mispositioning or an indirect effect not resolved","Single lab finding"]},{"year":2021,"claim":"Revealing that CAMSAP3 mutation activates mechanosensors YAP and PIEZO1 and causes proximal convoluted tubule cystogenesis established that noncentrosomal microtubule loss alters cell mechanics and mechanotransduction.","evidence":"Camsap3 mutant mice, YAP/PIEZO1 immunostaining, electron microscopy, proliferation assays","pmids":["33712686"],"confidence":"Medium","gaps":["Whether microtubule-dependent cortical stiffness change directly activates PIEZO1 not shown","Cyst progression and reversibility not characterized"]},{"year":2021,"claim":"Demonstrating that CAMSAP3 KO in lung cancer cells causes senescence-associated phenotypes through ERK/cyclin D1 downregulation, with vimentin scaffolding a CAMSAP3-ERK complex, identified a signaling axis linking microtubule organization to cell cycle control.","evidence":"CRISPR KO, IP/mass spectrometry, immunoblotting, flow cytometry, xenograft model","pmids":["34724356"],"confidence":"Medium","gaps":["Direct binding between CAMSAP3 and ERK vs vimentin-mediated proximity not resolved","Cancer-type specificity not addressed"]},{"year":2022,"claim":"Extending the central pair phenotype to vestibular hair cell kinocilia confirmed that CAMSAP3-dependent central pair formation is a generalizable mechanism across 9+2 axonemes.","evidence":"Conditional KO mouse, TEM, immunofluorescence, behavioral assays","pmids":["35783105"],"confidence":"Medium","gaps":["Whether kinocilium shortening is due to central pair loss or independent function not delineated","Behavioral phenotype not extensively characterized"]},{"year":2023,"claim":"Identification of paracingulin/CGNL1 as the tight junction receptor that recruits CAMSAP3 via coiled-coil interactions through ZO-1 answered how CAMSAP3 is targeted to cell-cell junctions and linked junctional microtubules to epithelial barrier architecture.","evidence":"GST pulldown, KO cell lines, ultrastructure expansion microscopy, in vivo mouse intestinal epithelium","pmids":["37013686"],"confidence":"High","gaps":["Whether CGNL1 is the sole junctional receptor or additional anchors exist not excluded","Functional consequence on barrier permeability not quantified"]},{"year":2024,"claim":"Biochemical reconstitution showing that CAMSAP3 dimerizes through its C-terminal α-helix to enhance CKK domain minus-end binding provided the first molecular-level explanation for how CAMSAP3 achieves high-affinity minus-end decoration and dynamic regulation.","evidence":"In vitro dimerization assays, microtubule-binding assays, domain mutagenesis, live cell imaging","pmids":["39479887"],"confidence":"High","gaps":["Crystal or cryo-EM structure of the dimer not available","Whether dimerization is regulated in vivo (e.g., by phosphorylation) unknown"]},{"year":2024,"claim":"Showing that CAMSAP3 loss drives centrosomal microtubule clustering, impairs endosomal-lysosomal trafficking of EGFR, and confers osimertinib resistance — reversed by CAMSAP3 re-expression — provided a mechanistic link between noncentrosomal microtubule loss and drug resistance in NSCLC.","evidence":"siRNA/CRISPR depletion and overexpression, EGFR trafficking assays, xenograft model","pmids":["41381443"],"confidence":"Medium","gaps":["Generalizability to other receptor tyrosine kinases not tested","Patient-level validation of CAMSAP3 loss in osimertinib resistance lacking"]},{"year":2024,"claim":"Demonstrating that CAMSAP3 deficiency in keratinocytes shifts microtubules to centrosomal configuration and impairs both cell cycle progression and adherens/tight junction stability established that CAMSAP3-dependent noncentrosomal arrays are required for proliferation and intercellular adhesion in stratified epithelia.","evidence":"Inducible KO and rescue in HaCaT cells, immunofluorescence, flow cytometry","pmids":["38190868"],"confidence":"Medium","gaps":["In vivo skin phenotype not characterized","Whether other CAMSAP family members compensate in keratinocytes not addressed"]},{"year":null,"claim":"Key unresolved questions include the structural basis of CAMSAP3 dimerization and CKK minus-end recognition, the mechanism by which CAMSAP3 nucleates or stabilizes the central microtubule pair at the axonemal basal plate, and how CAMSAP3 minus-end occupancy suppresses αTAT1-dependent acetylation at the molecular level.","evidence":"","pmids":[],"confidence":"High","gaps":["No high-resolution structure of CAMSAP3 dimer or CKK-microtubule complex","Basal plate cofactors required for central pair formation unidentified","Molecular mechanism linking minus-end binding to αTAT1 exclusion unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,3,17]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,15]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1,17]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[8,9,10,21]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,15]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,18]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[8,9,10,21]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[15,20]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,11,13]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,5,19]}],"complexes":["CAMSAP3–ACF7/MACF1 minus-end anchoring complex"],"partners":["MACF1","CGNL1","CDH23","DYNC1H1","KATNA1","VIM","NCL","HMGB1"],"other_free_text":[]},"mechanistic_narrative":"CAMSAP3 is a microtubule minus-end binding protein that stabilizes noncentrosomal microtubule arrays and orients apico-basal polarity in diverse epithelial tissues, neurons, and ciliated cells. It dimerizes through a C-terminal α-helix that enhances CKK domain minus-end binding affinity, and its MBD and CKK domains cooperate to decorate and dynamically regulate minus ends; its CC1 domain directs apical cortical localization, where the spectraplakin ACF7/MACF1 anchors CAMSAP3-decorated minus ends to actin, and paracingulin/CGNL1 recruits CAMSAP3 to tight junctions [PMID:39479887, PMID:27802168, PMID:27693509, PMID:37013686]. In motile cilia and kinocilia, CAMSAP3 concentrates at the axonemal basal plate and is required for central microtubule pair formation; its loss produces primary ciliary dyskinesia-like phenotypes with disorganized ciliary beating [PMID:32482850, PMID:34319756, PMID:35783105]. CAMSAP3 suppresses αTAT1-dependent tubulin acetylation to maintain neuronal polarity and epithelial identity, and its loss deregulates Akt, ERK/cyclin D1, and mTORC1 signaling, disrupts endosomal-lysosomal trafficking, activates mechanosensors YAP/PIEZO1, and promotes epithelial-mesenchymal transition [PMID:30190432, PMID:30282632, PMID:33462112, PMID:33712686, PMID:41381443]."},"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 1","ntpm":59.9}],"url":"https://www.proteinatlas.org/search/CAMSAP3"},"hgnc":{"alias_symbol":["Nezha","PPP1R80"],"prev_symbol":["KIAA1543"]},"alphafold":{"accession":"Q9P1Y5","domains":[{"cath_id":"-","chopping":"18-155","consensus_level":"medium","plddt":91.7551,"start":18,"end":155},{"cath_id":"1.10.418.10","chopping":"161-189_220-326","consensus_level":"medium","plddt":90.2585,"start":161,"end":326},{"cath_id":"3.10.20.360","chopping":"1122-1234","consensus_level":"high","plddt":90.3873,"start":1122,"end":1234}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P1Y5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P1Y5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P1Y5-F1-predicted_aligned_error_v6.png","plddt_mean":59.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CAMSAP3","jax_strain_url":"https://www.jax.org/strain/search?query=CAMSAP3"},"sequence":{"accession":"Q9P1Y5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9P1Y5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9P1Y5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P1Y5"}},"corpus_meta":[{"pmid":"23169647","id":"PMC_23169647","title":"Nezha/CAMSAP3 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katanin.","date":"2017","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/28386021","citation_count":18,"is_preprint":false},{"pmid":"37013686","id":"PMC_37013686","title":"Paracingulin recruits CAMSAP3 to tight junctions and regulates microtubule and polarized epithelial cell organization.","date":"2023","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/37013686","citation_count":16,"is_preprint":false},{"pmid":"27349180","id":"PMC_27349180","title":"Cadherin 23-C Regulates Microtubule Networks by Modifying CAMSAP3's Function.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27349180","citation_count":14,"is_preprint":false},{"pmid":"37019300","id":"PMC_37019300","title":"CAMSAP3 negatively regulates lung cancer cell invasion and angiogenesis through nucleolin/HIF-1α mRNA complex stabilization.","date":"2023","source":"Life 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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":"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. 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depletion of both CAMSAPs caused marked reduction of microtubules with polymerizing plus ends and perturbed early endosome and Golgi distribution.\",\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 — clean KD with defined cellular phenotypes, replicated across two CAMSAP family members, highly cited foundational study\",\n      \"pmids\": [\"23169647\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CAMSAP3 accumulates at apical cortices of polarized intestinal epithelial cells and tethers longitudinal microtubule minus ends there, orienting microtubules along the apical-to-basal axis; its CC1 domain is required for apical localization, and forced mislocalization disrupts epithelial architecture.\",\n      \"method\": \"Camsap3 mutation/depletion in mouse intestinal cells and Caco-2 cells, domain mutagenesis, immunofluorescence, forced mislocalization experiments\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined phenotype plus domain mutagenesis, replicated in two cell systems\",\n      \"pmids\": [\"26715742\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ACF7 (MACF1), a spectraplakin family cytoskeletal crosslinking protein, specifically binds CAMSAP3 and is required for apical localization of CAMSAP3-decorated microtubule minus ends in intestinal epithelial cells; CAMSAP3 knockout (but not CAMSAP2) caused microtubule reorganization into a centrosomal radial array and redistribution of Rab11-positive endosomes away from the apical surface.\",\n      \"method\": \"Reciprocal Co-IP, knockout cell lines, immunofluorescence, 3D cyst culture\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP plus KO with multiple defined phenotypes\",\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; this CAMSAP3-ACF7 complex couples microtubule and actin dynamics to regulate retrograde flow, focal adhesion size, and cell migration in Caco-2 epithelial cells.\",\n      \"method\": \"Co-IP, pulldown, live imaging, siRNA depletion, focal adhesion and migration assays\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus loss-of-function with multiple orthogonal phenotypic readouts\",\n      \"pmids\": [\"27693509\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CAMSAP3 accumulates in the pericentrosomal area and cooperates with dynein and katanin to mediate microtubule detachment from the centrosome; depletion of CAMSAP3 prevents microtubule release and causes focusing of microtubules at centrosomes.\",\n      \"method\": \"siRNA depletion, immunofluorescence, live imaging, Co-IP with dynein and katanin\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined phenotype and Co-IP with molecular partners, single lab\",\n      \"pmids\": [\"28386021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CAMSAP3 interacts with CG-NAP and regulates Golgi vesicle translocation in epithelial cells; depletion of either CAMSAP3 or CG-NAP causes Golgi membrane fragmentation, and stathmin-dependent microtubule dynamics graded along the radial axis contributes to perinuclear Golgi assembly.\",\n      \"method\": \"Co-IP, siRNA depletion, immunofluorescence, Golgi morphology assays\",\n      \"journal\": \"Journal of genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus loss-of-function with defined Golgi phenotype, single lab\",\n      \"pmids\": [\"28089391\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CAMSAP3 is enriched in axons of hippocampal neurons and preferentially localizes to less-acetylated microtubules; CAMSAP3 mutation causes supernumerary axons and increased tubulin acetylation; CAMSAP3 depletion promotes αTAT1-dependent tubulin acetylation, and αTAT1 depletion abolishes multi-axon formation caused by CAMSAP3 loss, placing CAMSAP3 upstream of αTAT1 in axonal polarity maintenance.\",\n      \"method\": \"Genetic epistasis (double KD), mouse mutants, immunofluorescence, nocodazole resistance assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — epistasis experiment plus loss-of-function in primary neurons with multiple defined phenotypes\",\n      \"pmids\": [\"30190432\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Loss of CAMSAP3 in lung carcinoma cells increases tubulin acetylation, which upregulates Akt activity and promotes EMT at the transcriptional level; CAMSAP3 thus protects epithelial phenotype by suppressing Akt activity via microtubule regulation.\",\n      \"method\": \"CAMSAP3 deletion cell lines, Akt activity assays, tubulin acetylation immunoblot, EMT marker analysis\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO with mechanistic pathway placement, single lab\",\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; its loss results in absence of the central microtubule pair in most cilia and disorganized basal body polarity, causing defective synchronized ciliary beating and PCD-like phenotypes; confirmed in Xenopus by morpholino knockdown.\",\n      \"method\": \"Hypomorphic knockdown mouse model, Xenopus morpholino, electron microscopy, immunofluorescence, 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 — loss-of-function in two model organisms with ultrastructural and functional validation\",\n      \"pmids\": [\"32482850\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3 concentrates at the axonemal basal plate (transition zone) where the central microtubule pair initiates; CAMSAP3 dysfunction causes loss of the central pair and partial distortion of the basal plate, as well as failure of coordinated multicilia beating in tracheal airway epithelial cells.\",\n      \"method\": \"Mouse mutants, super-resolution immunofluorescence, electron microscopy, ciliary beat analysis\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ultrastructural localization plus loss-of-function with defined structural and functional phenotypes\",\n      \"pmids\": [\"34319756\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3 localizes to the base of cilia in a polarized manner in oviduct multiciliated cells; its mutation disrupts intracellular coordination of basal body orientation and the assembly of microtubules interconnecting basal bodies, without affecting planar cell polarity factor localization, indicating a distinct mechanism from CELSR1.\",\n      \"method\": \"Mouse mutants, immunofluorescence, super-resolution microscopy, basal body orientation quantification\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis/pathway separation plus loss-of-function with structural phenotype\",\n      \"pmids\": [\"33468623\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3 concentrates at apical domains of differentiating ependymal cells and generates apical MT networks that support lysosomal positioning; CAMSAP3 mutation downregulates mTORC1 activity, reduces apical lysosome distribution, and impairs ependymal cell apical domain expansion, leading to abnormal lateral ventricle narrowing.\",\n      \"method\": \"Camsap3 mutant mice, mTORC1 activity assays, lysosome immunofluorescence, ventricle morphometry\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function in mouse with mechanistic pathway placement, single lab\",\n      \"pmids\": [\"33462112\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CAMSAP3-mediated microtubule networks are required to maintain mechanical properties of proximal convoluted tubule (PCT) cells; CAMSAP3 mutation activates mechanosensors YAP and PIEZO1, induces cell flattening and proliferation, and causes PCT cyst formation.\",\n      \"method\": \"Camsap3 mutant mice, YAP/PIEZO1 immunostaining, electron microscopy, proliferation assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mouse loss-of-function with mechanosensor pathway placement, 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 arrest via downregulation of phospho-ERK and cyclin D1; vimentin acts as a scaffold for the CAMSAP3-ERK signaling complex, identified by immunoprecipitation/mass spectrometry.\",\n      \"method\": \"CRISPR-Cas9 KO, IP/mass spectrometry, immunoblotting, flow cytometry, xenograft model\",\n      \"journal\": \"Cancer medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO with mechanistic pathway placement and proteomic partner identification, single lab\",\n      \"pmids\": [\"34724356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The C isoform of CDH23 directly binds the CKK domain of CAMSAP3 via a conserved N-terminal CKK-binding motif (CBM) and inhibits CAMSAP3-induced microtubule bundle formation; the Usher Syndrome 1D mutation CDH23 R3175H (mouse R55H) reduces this interaction both in vitro and in vivo.\",\n      \"method\": \"Co-IP, pulldown, in vitro binding assays, mutagenesis, in vivo mouse models\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — direct binding with mutagenesis and in vivo validation\",\n      \"pmids\": [\"27349180\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Paracingulin (CGNL1), but not PLEKHA7, interacts with CAMSAP3 via their coiled-coil regions (GST pulldown) and recruits CAMSAP3 to tight junctions via the ZO-1-associated pool; CGNL1 knockout causes loss of junctional CAMSAP3, disorganized cytoplasmic microtubules, and disrupted epithelial architecture.\",\n      \"method\": \"GST pulldown, KO cell lines, ultrastructure expansion microscopy, in vivo mouse intestinal epithelium\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — GST pulldown defines interaction domain plus KO with multiple orthogonal phenotypes in vitro and in vivo\",\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; CAMSAP3 knockout increases HIF-1α expression and downstream VEGFA and MMP2/9, promoting invasion and angiogenesis; identified by proteomic analysis and RNA immunoprecipitation.\",\n      \"method\": \"CRISPR-Cas9 KO, IP/mass spectrometry, RNA immunoprecipitation, in vitro invasion/angiogenesis assays, in vivo xenograft\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — proteomic interaction plus RNA-IP with loss-of-function, 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; dimerization enhances the microtubule-binding affinity of the CKK domain and enables the CKK domain to regulate minus-end dynamics; the combined action of the microtubule-binding domain (MBD) and C-terminal α-helix mediates minus-end decoration and dynamic regulation.\",\n      \"method\": \"Biochemical dimerization assays, in vitro microtubule-binding assays, domain mutagenesis, live cell imaging\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with domain mutagenesis defining molecular mechanism\",\n      \"pmids\": [\"39479887\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 interacts with acetylated HMGB1 in the cytoplasm; TSA-induced HMGB1 acetylation drives HMGB1 cytoplasmic translocation and secretion promoting autophagic cell death, a process that requires CAMSAP3; CAMSAP3 knockout abolishes this TSA-mediated autophagic cell death.\",\n      \"method\": \"CRISPR-Cas9 KO, proteomic analysis, immunoprecipitation, immunofluorescence, autophagic detection assays\",\n      \"journal\": \"Biochimica et biophysica acta. General subjects\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus KO with defined functional outcome, single lab\",\n      \"pmids\": [\"38598971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 depletion in NSCLC cells causes centrosomal microtubule clustering, impairs the endosomal-lysosomal system, and drives EGFR translocation to the perinuclear MTOC, blocking plasma membrane recycling and promoting lysosomal EGFR degradation, thereby conferring osimertinib resistance; CAMSAP3 overexpression in resistant cells restores microtubule organization and drug sensitivity.\",\n      \"method\": \"siRNA/CRISPR depletion, CAMSAP3 overexpression, EGFR trafficking assays, xenograft model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — bidirectional genetic manipulation with defined mechanistic pathway, single lab\",\n      \"pmids\": [\"41381443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CAMSAP3 deficiency in keratinocytes shifts microtubules from non-centrosomal to centrosomal configuration, causes cell cycle exit, delayed cytokinesis, and impairs formation and stability of adherens junctions and tight junctions; re-expression of CAMSAP3 rescues these defects.\",\n      \"method\": \"Inducible CAMSAP3 KO and re-expression in HaCaT cells, immunofluorescence, flow cytometry\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — bidirectional genetic rescue experiment with multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"38190868\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CAMSAP3 is required for formation and/or maintenance of the central microtubule pair in vestibular hair cell kinocilia; conditional knockout of CAMSAP3 results in shorter kinocilia and more frequent absence of the central MT pair, linking CAMSAP3 to axoneme length and stability.\",\n      \"method\": \"Conditional KO mouse model, immunofluorescence, transmission electron microscopy, behavioral assays\",\n      \"journal\": \"Frontiers in cellular neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with ultrastructural phenotype, single lab\",\n      \"pmids\": [\"35783105\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CAMSAP3-mediated microtubules are required for maintenance of transzonal projections (TZPs) between granulosa cells and oocytes in ovarian follicles; CAMSAP3 KO mice are infertile with fewer late-stage follicles, reduced TZP number, and disorganized microtubules in TZPs; CAMSAP3 also modulates TZP morphology by organizing both microtubules and F-actin.\",\n      \"method\": \"CAMSAP3 KO mice, super-resolution microscopy, TZP quantification, follicle staging\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO with defined ultrastructural and fertility phenotypes; preprint, not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.09.26.678897\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"CAMSAP3 is a microtubule minus-end binding protein that forms dimers via its C-terminal α-helix (enhancing CKK domain binding affinity) and stabilizes noncentrosomal microtubule minus ends through its MBD and CKK domains; it localizes to apical cortices, pericentrosomal regions, tight junctions (via paracingulin/CGNL1), and the axonemal basal plate, where it cooperates with ACF7/MACF1 to anchor minus ends to actin, with dynein and katanin to release microtubules from the centrosome, and with CDH23-C to modulate microtubule bundling, thereby orienting apico-basal microtubule polarity in epithelia, maintaining neuronal polarity by suppressing αTAT1-dependent tubulin acetylation, forming the central microtubule pair in motile cilia and kinocilia, and regulating downstream signaling through Akt, ERK/cyclin D1, and mTORC1 pathways.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"CAMSAP3 is a microtubule minus-end binding protein that stabilizes noncentrosomal microtubule arrays and orients apico-basal polarity in diverse epithelial tissues, neurons, and ciliated cells. It dimerizes through a C-terminal α-helix that enhances CKK domain minus-end binding affinity, and its MBD and CKK domains cooperate to decorate and dynamically regulate minus ends; its CC1 domain directs apical cortical localization, where the spectraplakin ACF7/MACF1 anchors CAMSAP3-decorated minus ends to actin, and paracingulin/CGNL1 recruits CAMSAP3 to tight junctions [PMID:39479887, PMID:27802168, PMID:27693509, PMID:37013686]. In motile cilia and kinocilia, CAMSAP3 concentrates at the axonemal basal plate and is required for central microtubule pair formation; its loss produces primary ciliary dyskinesia-like phenotypes with disorganized ciliary beating [PMID:32482850, PMID:34319756, PMID:35783105]. CAMSAP3 suppresses αTAT1-dependent tubulin acetylation to maintain neuronal polarity and epithelial identity, and its loss deregulates Akt, ERK/cyclin D1, and mTORC1 signaling, disrupts endosomal-lysosomal trafficking, activates mechanosensors YAP/PIEZO1, and promotes epithelial-mesenchymal transition [PMID:30190432, PMID:30282632, PMID:33462112, PMID:33712686, PMID:41381443].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Establishing that CAMSAP3 and CAMSAP2 specifically recognize and stabilize noncentrosomal microtubule minus ends answered the fundamental question of how epithelial cells maintain acentrosomal microtubule networks and organize endomembranes.\",\n      \"evidence\": \"siRNA depletion with immunofluorescence and live imaging in epithelial cells\",\n      \"pmids\": [\"23169647\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of minus-end recognition not defined at the molecular level\", \"Relative contributions of CAMSAP2 vs CAMSAP3 not separated\", \"How microtubule stabilization suppresses centrosomal organization unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrating that CAMSAP3 accumulates at apical cortices via its CC1 domain and tethers minus ends there established the molecular basis of apico-basal microtubule orientation in intestinal epithelia.\",\n      \"evidence\": \"Domain mutagenesis and forced mislocalization in mouse intestinal cells and Caco-2 cells\",\n      \"pmids\": [\"26715742\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the apical receptor for the CC1 domain unknown\", \"Whether CC1-mediated localization is conserved across all polarized epithelia not tested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of ACF7/MACF1 as a direct CAMSAP3 partner that anchors minus-end-decorated microtubules to actin filaments resolved how the noncentrosomal microtubule network is physically connected to the cortical cytoskeleton and influences focal adhesion dynamics and migration.\",\n      \"evidence\": \"Reciprocal Co-IP, pulldown, KO cell lines, live imaging, focal adhesion and migration assays in Caco-2 cells and 3D cysts\",\n      \"pmids\": [\"27802168\", \"27693509\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of CAMSAP3–ACF7 interaction not mapped to specific domains\", \"Contribution of other spectraplakins not excluded\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Discovery that CDH23-C directly binds the CKK domain via a conserved CBM motif and inhibits CAMSAP3-induced microtubule bundling linked CAMSAP3 to stereocilia/inner ear biology and the Usher Syndrome 1D mutation R3175H.\",\n      \"evidence\": \"Co-IP, in vitro pulldown, mutagenesis, in vivo mouse models\",\n      \"pmids\": [\"27349180\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological consequence of CDH23–CAMSAP3 disruption on hearing not directly shown\", \"Whether other CKK-binding partners compete with CDH23 unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of CAMSAP3 interaction with CG-NAP and its role in Golgi vesicle translocation provided the first link between CAMSAP3-organized microtubules and Golgi ribbon integrity.\",\n      \"evidence\": \"Co-IP, siRNA depletion, Golgi morphology assays in epithelial cells\",\n      \"pmids\": [\"28089391\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding interface between CAMSAP3 and CG-NAP not mapped\", \"Golgi phenotype not validated in vivo\", \"Whether CAMSAP2 compensates at the Golgi not tested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showing that CAMSAP3 cooperates with dynein and katanin at the pericentrosomal area to release microtubules from centrosomes answered how noncentrosomal microtubules are generated from centrosomal precursors.\",\n      \"evidence\": \"siRNA depletion, live imaging, Co-IP with dynein and katanin\",\n      \"pmids\": [\"28386021\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Order of recruitment (CAMSAP3 vs dynein vs katanin) not resolved\", \"Reconstitution of microtubule release in vitro not achieved\", \"Single lab finding\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Genetic epistasis placing CAMSAP3 upstream of αTAT1-dependent tubulin acetylation in maintaining single-axon polarity established a new regulatory axis: CAMSAP3 suppresses tubulin acetylation to restrict axon number.\",\n      \"evidence\": \"Double knockdown epistasis, mouse Camsap3 mutants, nocodazole resistance assays in hippocampal neurons\",\n      \"pmids\": [\"30190432\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism by which CAMSAP3 inhibits αTAT1 access not defined\", \"Whether this axis operates in other neuronal subtypes not tested\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Linking CAMSAP3 loss to increased tubulin acetylation, Akt activation, and EMT in lung carcinoma extended the tubulin acetylation axis to signaling and cancer biology, showing CAMSAP3 protects epithelial identity.\",\n      \"evidence\": \"CAMSAP3 deletion cell lines, Akt activity assays, EMT marker analysis\",\n      \"pmids\": [\"30282632\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanism connecting tubulin acetylation to Akt activation not delineated\", \"In vivo tumor progression data lacking at this point\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrating that CAMSAP3 localizes to axonemal bases and is required for central microtubule pair formation in motile cilia revealed an unexpected structural role, explaining PCD-like phenotypes upon CAMSAP3 loss.\",\n      \"evidence\": \"Hypomorphic knockdown mouse, Xenopus morpholino, electron microscopy, ciliary beat analysis\",\n      \"pmids\": [\"32482850\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CAMSAP3 nucleates or stabilizes the central pair specifically is unknown\", \"Whether CAMSAP3 acts alone or requires cofactors at the basal plate not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Refining CAMSAP3 localization to the basal plate (transition zone) and demonstrating that its loss distorts basal plate ultrastructure and disrupts basal body coordination in multiciliated airways and oviduct cells consolidated the ciliary role and separated it mechanistically from planar cell polarity signaling via CELSR1.\",\n      \"evidence\": \"Mouse mutants, super-resolution microscopy, electron microscopy, basal body orientation quantification in tracheal and oviduct cells\",\n      \"pmids\": [\"34319756\", \"33468623\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Basal plate binding partners of CAMSAP3 not identified\", \"Whether CAMSAP3 functions at primary (non-motile) cilia not tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connecting CAMSAP3-organized microtubules to lysosomal positioning, mTORC1 signaling, and ependymal cell apical domain expansion broadened the downstream effector repertoire beyond acetylation-Akt, and showed tissue-level consequences (ventricle narrowing) of CAMSAP3 loss.\",\n      \"evidence\": \"Camsap3 mutant mice, mTORC1 activity assays, lysosome immunofluorescence, ventricle morphometry\",\n      \"pmids\": [\"33462112\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether mTORC1 downregulation is a direct consequence of lysosome mispositioning or an indirect effect not resolved\", \"Single lab finding\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Revealing that CAMSAP3 mutation activates mechanosensors YAP and PIEZO1 and causes proximal convoluted tubule cystogenesis established that noncentrosomal microtubule loss alters cell mechanics and mechanotransduction.\",\n      \"evidence\": \"Camsap3 mutant mice, YAP/PIEZO1 immunostaining, electron microscopy, proliferation assays\",\n      \"pmids\": [\"33712686\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether microtubule-dependent cortical stiffness change directly activates PIEZO1 not shown\", \"Cyst progression and reversibility not characterized\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrating that CAMSAP3 KO in lung cancer cells causes senescence-associated phenotypes through ERK/cyclin D1 downregulation, with vimentin scaffolding a CAMSAP3-ERK complex, identified a signaling axis linking microtubule organization to cell cycle control.\",\n      \"evidence\": \"CRISPR KO, IP/mass spectrometry, immunoblotting, flow cytometry, xenograft model\",\n      \"pmids\": [\"34724356\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding between CAMSAP3 and ERK vs vimentin-mediated proximity not resolved\", \"Cancer-type specificity not addressed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Extending the central pair phenotype to vestibular hair cell kinocilia confirmed that CAMSAP3-dependent central pair formation is a generalizable mechanism across 9+2 axonemes.\",\n      \"evidence\": \"Conditional KO mouse, TEM, immunofluorescence, behavioral assays\",\n      \"pmids\": [\"35783105\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether kinocilium shortening is due to central pair loss or independent function not delineated\", \"Behavioral phenotype not extensively characterized\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of paracingulin/CGNL1 as the tight junction receptor that recruits CAMSAP3 via coiled-coil interactions through ZO-1 answered how CAMSAP3 is targeted to cell-cell junctions and linked junctional microtubules to epithelial barrier architecture.\",\n      \"evidence\": \"GST pulldown, KO cell lines, ultrastructure expansion microscopy, in vivo mouse intestinal epithelium\",\n      \"pmids\": [\"37013686\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CGNL1 is the sole junctional receptor or additional anchors exist not excluded\", \"Functional consequence on barrier permeability not quantified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Biochemical reconstitution showing that CAMSAP3 dimerizes through its C-terminal α-helix to enhance CKK domain minus-end binding provided the first molecular-level explanation for how CAMSAP3 achieves high-affinity minus-end decoration and dynamic regulation.\",\n      \"evidence\": \"In vitro dimerization assays, microtubule-binding assays, domain mutagenesis, live cell imaging\",\n      \"pmids\": [\"39479887\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Crystal or cryo-EM structure of the dimer not available\", \"Whether dimerization is regulated in vivo (e.g., by phosphorylation) unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showing that CAMSAP3 loss drives centrosomal microtubule clustering, impairs endosomal-lysosomal trafficking of EGFR, and confers osimertinib resistance — reversed by CAMSAP3 re-expression — provided a mechanistic link between noncentrosomal microtubule loss and drug resistance in NSCLC.\",\n      \"evidence\": \"siRNA/CRISPR depletion and overexpression, EGFR trafficking assays, xenograft model\",\n      \"pmids\": [\"41381443\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Generalizability to other receptor tyrosine kinases not tested\", \"Patient-level validation of CAMSAP3 loss in osimertinib resistance lacking\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrating that CAMSAP3 deficiency in keratinocytes shifts microtubules to centrosomal configuration and impairs both cell cycle progression and adherens/tight junction stability established that CAMSAP3-dependent noncentrosomal arrays are required for proliferation and intercellular adhesion in stratified epithelia.\",\n      \"evidence\": \"Inducible KO and rescue in HaCaT cells, immunofluorescence, flow cytometry\",\n      \"pmids\": [\"38190868\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo skin phenotype not characterized\", \"Whether other CAMSAP family members compensate in keratinocytes not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis of CAMSAP3 dimerization and CKK minus-end recognition, the mechanism by which CAMSAP3 nucleates or stabilizes the central microtubule pair at the axonemal basal plate, and how CAMSAP3 minus-end occupancy suppresses αTAT1-dependent acetylation at the molecular level.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No high-resolution structure of CAMSAP3 dimer or CKK-microtubule complex\", \"Basal plate cofactors required for central pair formation unidentified\", \"Molecular mechanism linking minus-end binding to αTAT1 exclusion unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 3, 17]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 17]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [8, 9, 10, 21]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 15]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 18]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [8, 9, 10, 21]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [15, 20]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 11, 13]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 5, 19]}\n    ],\n    \"complexes\": [\n      \"CAMSAP3–ACF7/MACF1 minus-end anchoring complex\"\n    ],\n    \"partners\": [\n      \"MACF1\",\n      \"CGNL1\",\n      \"CDH23\",\n      \"DYNC1H1\",\n      \"KATNA1\",\n      \"VIM\",\n      \"NCL\",\n      \"HMGB1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}