{"gene":"AMOTL2","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2007,"finding":"Amotl2 interacts preferentially with phosphorylated c-Src and facilitates its outward translocation to the membrane, regulating membrane architecture and F-actin organization; knockdown of amotl2 in zebrafish delays epiboly and impairs convergence/extension cell movements, and amotl2-deficient cells fail to migrate properly with loss of membrane protrusions.","method":"Zebrafish morpholino knockdown, mosaic transplantation, co-immunoprecipitation, co-localization with endosomal markers (RhoB, EEA1)","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP and in vivo loss-of-function in zebrafish with defined cellular phenotypes, single lab","pmids":["17293535"],"is_preprint":false},{"year":2011,"finding":"Amotl2 promotes MAPK/ERK activation via c-Src in endothelial cells, dependent on phosphorylation of tyrosine residue at position 103 but independent of the C-terminal PDZ-binding domain; knockdown impairs endothelial cell proliferation, migration, polarity, and tube formation in vitro, and intersegmental vessel growth in zebrafish.","method":"Zebrafish transgenic line knockdown, HUVEC siRNA knockdown, site-directed mutagenesis (Y103), ERK activity assays, cell polarity and tube formation assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — mutagenesis combined with in vitro biochemical assays and in vivo zebrafish validation, two orthogonal model systems","pmids":["21937427"],"is_preprint":false},{"year":2012,"finding":"Amotl2 attenuates Wnt/β-catenin signaling by associating with and trapping β-catenin in Rab11-positive recycling endosomes, reducing the amount of β-catenin available in the cytosol and nucleus; knockdown in zebrafish causes embryonic dorsalization rescued by co-knockdown of β-catenin2.","method":"Zebrafish morpholino knockdown, genetic epistasis with axin1 mutant (masterblind), co-immunoprecipitation, immunofluorescence with Rab11 co-localization in mammalian cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, co-localization with Rab11 endosomes, in vivo epistasis in zebrafish, replicated across cell and animal models","pmids":["22362771"],"is_preprint":false},{"year":2013,"finding":"AMOTL2 co-immunoprecipitates with TAZ via the WW domain of TAZ and the PPXY motif in the N-terminus of AMOTL2; AMOTL2 co-localizes with TAZ in the cytoplasm and regulates TAZ cytoplasm-to-nucleus translocation through direct protein-protein interaction, inhibiting TAZ-dependent transcription of surfactant genes in lung cells in a Hippo-independent manner.","method":"Co-immunoprecipitation, immunofluorescence co-localization, luciferase reporter assay, domain mutagenesis (PPXY motif), overexpression in H441 lung cells","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain mapping and reporter assay, single lab, single cell type","pmids":["23911299"],"is_preprint":false},{"year":2013,"finding":"Amotl2 interacts with the scaffold protein LL5β at synaptic podosomes in myotubes and neuromuscular junctions in vivo; depletion of Amotl2 in myotubes increases size of synaptic podosomes and alters postsynaptic topology, and depletion in fibroblasts disrupts invadopodia.","method":"Affinity purification/mass spectrometry (LL5β-associated proteins), co-immunoprecipitation, immunofluorescence in vivo and in vitro, siRNA knockdown with morphometric analysis","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-based identification followed by co-IP and loss-of-function with defined phenotype, single lab","pmids":["23525008"],"is_preprint":false},{"year":2014,"finding":"AmotL2 associates with the VE-cadherin adherens junction complex and couples it to contractile actin fibres; inactivation of amotL2 in zebrafish, mouse, and endothelial cell culture dissociates VE-cadherin from cytoskeletal tensile forces, impairing aortic vessel lumen expansion.","method":"Gene inactivation in zebrafish and mouse, endothelial cell culture knockdown, co-immunoprecipitation with VE-cadherin complex, traction force microscopy, live imaging","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal model systems (zebrafish, mouse, cell culture), co-IP with junctional complex, functional mechanical readouts across independent labs","pmids":["24806444"],"is_preprint":false},{"year":2014,"finding":"Hypoxic stress induces c-Fos-dependent expression of a p60 AmotL2 isoform; p60 AmotL2 interacts with the Crb3 and Par3 polarity complexes, retaining them in large vesicles and preventing them from reaching the apical membrane, causing loss of apical-basal polarity and promoting tumor invasion.","method":"Immunoprecipitation, immunofluorescence, siRNA knockdown in vitro, mouse xenograft in vivo, c-Fos promoter analysis, vesicle trafficking assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP with polarity complexes, mechanistic promoter analysis, in vitro and in vivo validation with defined cellular phenotype","pmids":["25080976"],"is_preprint":false},{"year":2015,"finding":"mTORC2 phosphorylates AMOTL2 at serine 760; phosphomimetic S760E mutation blocks AMOTL2's ability to bind and repress YAP, increasing YAP target gene expression, foci formation, and metastatic properties, whereas non-phosphorylatable S760A mutant retains YAP repression activity in glioblastoma cells and xenografts.","method":"Gene-trap in mouse glioma model, phosphosite mutagenesis (S760A/S760E), co-immunoprecipitation with YAP, YAP transcriptional reporter assays, foci formation assay, xenograft tumor growth","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — site-directed mutagenesis with biochemical and in vivo validation, multiple orthogonal phenotypic readouts, in vitro and in vivo","pmids":["25998128"],"is_preprint":false},{"year":2017,"finding":"Par3 is essential for localization of AmotL2 to cellular junctions, where it associates with VE/E-cadherin to organize radial actin filaments; loss of this Par3-AmotL2-cadherin-actin axis impairs aortic lumen expansion and epithelial hexagonal packing.","method":"Gene inactivation in zebrafish, siRNA knockdown in cell culture, immunofluorescence, co-immunoprecipitation","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP and in vivo genetic epistasis in zebrafish, single lab, two orthogonal systems","pmids":["28790366"],"is_preprint":false},{"year":2017,"finding":"p100 AmotL2 forms a complex with E-cadherin that associates with radial actin filaments connecting cells across multiple layers; genetic inactivation of amotL2 leads to loss of contractile actin filaments and perturbed epithelial packing geometry; amotL2 is required for blastocyst hatching in mouse and zebrafish via tension generation, phenocopied by myosin II inhibitor blebbistatin.","method":"Co-immunoprecipitation, zebrafish and mouse genetic inactivation, immunofluorescence, blebbistatin pharmacological inhibition, blastocyst hatching assay","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP, multiple genetic model systems (zebrafish, mouse), pharmacological inhibition corroborating mechanism, defined developmental phenotype","pmids":["28842668"],"is_preprint":false},{"year":2020,"finding":"AMOTL2 is a binding partner of PPP2R2A (a PP2A regulatory subunit) in NSCLC cells; AMOTL2 binds PPP2R2A in the cytoplasm, reducing its nuclear localization and thereby preventing PPP2R2A-mediated dephosphorylation of JUN at Thr239, which suppresses AP-1-driven cell proliferation.","method":"CRISPR/Cas9 loss-of-function screen, mass spectrometry, co-immunoprecipitation, GST pull-down, immunofluorescence, subcellular fractionation, phospho-JUN immunoblotting","journal":"Biochimica et biophysica acta. Molecular cell research","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal binding assays (MS, co-IP, GST pull-down), functional mechanistic follow-up with subcellular localization and phosphorylation readout","pmids":["32950569"],"is_preprint":false},{"year":2021,"finding":"E3 ubiquitin ligase WWP1 mono-ubiquitinates AMOTL2 at K347 and K408; mono-ubiquitinated AMOTL2 interacts with LATS2 and facilitates recruitment of SAV1, promoting YAP phosphorylation and cytoplasmic sequestration/degradation; this process is coupled to Crumbs polarity complex at cell junctions under high cell density conditions.","method":"In vitro ubiquitination assay, ubiquitination site mutagenesis (K347/K408), co-immunoprecipitation with LATS2/SAV1, YAP phosphorylation immunoblotting, cell density contact inhibition assays, immunofluorescence","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — in vitro ubiquitination with site mutagenesis plus co-IP and functional YAP phosphorylation readout, multiple orthogonal methods","pmids":["34404733"],"is_preprint":false},{"year":2021,"finding":"AMOTL2 directly binds β-catenin and regulates its nuclear translocation in glioma cells; knockdown of AMOTL2 promotes β-catenin nuclear localization and downstream Wnt target gene expression, enhancing glioma proliferation, migration, and invasion.","method":"Co-immunoprecipitation, immunofluorescence, immunoblotting, siRNA knockdown in glioma cell lines","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — co-IP and immunofluorescence with functional loss-of-function assays, single lab, consistent with prior Wnt pathway findings","pmids":["34036399"],"is_preprint":false},{"year":2021,"finding":"Loss of MAGI1 causes accumulation of E-cadherin and AMOTL2 and increased ROCK and p38 SAPK activities; rescue experiments show that AMOTL2 depletion or p38 inhibition reverses the increased tumorigenicity of MAGI1-deficient cells, placing AMOTL2 upstream of a ROCK/p38 stress pathway in luminal breast cancer.","method":"Genetic epistasis (double knockdown), ROCK/p38 inhibitor pharmacology, cell stiffness measurement, tumorigenicity assays in vitro","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via double-depletion and pharmacological rescue, single lab, functional phenotype defined","pmids":["33707576"],"is_preprint":false},{"year":2021,"finding":"AMOTL2 restrains YAP1 activation in airway smooth muscle cells; overexpression of AMOTL2 suppresses TGF-β1-induced YAP1 nuclear translocation, and reactivation of YAP1 reverses AMOTL2-mediated suppression of proliferation and ECM deposition.","method":"Overexpression and siRNA knockdown in ASM cells, YAP1 nuclear localization assay, proliferation and ECM deposition functional assays, rescue with constitutively active YAP1","journal":"Environmental toxicology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, overexpression/KD with functional readout and YAP1 rescue, no direct binding assay reported in abstract","pmids":["34323359"],"is_preprint":false},{"year":2022,"finding":"MEF2D binds the MEF2 cis-acting element in the upstream promoter region of AMOTL2, inhibiting its transcriptional expression, thereby activating YAP signaling and promoting HCC cell migration and proliferation.","method":"Dual luciferase reporter assay, ChIP (implied by promoter binding), overexpression experiments in hepatoma cells","journal":"International journal of clinical and experimental pathology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — reporter assay for promoter binding, single lab, limited mechanistic depth described in abstract","pmids":["35698637"],"is_preprint":false},{"year":2023,"finding":"CLK2 inhibition promotes alternative splicing of AMOTL2 producing an exon-skipped isoform that can no longer associate with membrane-bound proteins, resulting in decreased YAP phosphorylation and decreased membrane localization of YAP, thereby activating YAP-driven transcription.","method":"High-throughput chemical screen, RT-PCR for alternative splicing, YAP phosphorylation and localization assays, CLK2 inhibitor (SM04690) pharmacology","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chemical-genetic approach with splicing and functional YAP readouts, single lab, mechanism defined but no direct binding reconstitution","pmids":["38126343"],"is_preprint":false},{"year":2023,"finding":"p60 AmotL2 (expressed in invading tumor cells) binds to the p100 AmotL2 isoform and uncouples the mechanical constraint of radial actin filaments from E-cadherin; the E-cadherin/p100AmotL2 complex is directly connected to the nuclear membrane, and p60AmotL2 expression inactivates this connection, altering nuclear lamina properties and potentiating ameboid invasion through extracellular matrix micropores.","method":"Co-immunoprecipitation between isoforms, nuclear lamina connectivity assays, atomic force microscopy, micropore invasion assays, overexpression/knockdown in tumor cell lines","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP between isoforms plus functional invasion assay and biophysical nuclear stiffness measurement, single lab","pmids":["37443716"],"is_preprint":false},{"year":2023,"finding":"AmotL2 connects junctional VE-cadherin and actin filaments to the nuclear lamina in endothelial cells; AmotL2 is essential for radial actin filament formation and endothelial cell alignment in response to blood flow; loss of endothelial AmotL2 in mice causes a pro-inflammatory response and abdominal aortic aneurysms. Molecular analysis showed VE-cadherin binds AmotL2 and actin, transmitting extracellular mechanical signals to the nuclear membrane.","method":"Endothelial-specific mouse knockout, co-immunoprecipitation (VE-cadherin/AmotL2/actin), immunofluorescence for nuclear lamina connectivity, transcriptome analysis, live imaging","journal":"Nature cardiovascular research","confidence":"High","confidence_rationale":"Tier 2 / Strong — endothelial-specific KO mouse with defined vascular disease phenotype, co-IP demonstrating tripartite complex, nuclear lamina connectivity assay, transcriptome validation","pmids":["39195920"],"is_preprint":false},{"year":2024,"finding":"ARNTL2 negatively regulates AMOTL2 transcription by directly binding to the AMOTL2 promoter; reduced AMOTL2 decreases its recruitment and stabilization of LATS1/2 kinases, reducing LATS-dependent YAP phosphorylation and promoting YAP nuclear translocation and NPC invasion; inhibition of AMOTL2 counteracted the effect of ARNTL2 knockdown.","method":"ChIP (ARNTL2 binding to AMOTL2 promoter), siRNA/shRNA knockdown epistasis, LATS1/2 co-immunoprecipitation with AMOTL2, YAP phosphorylation and localization assays, xenograft mouse model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP for promoter binding, co-IP for LATS interaction, epistasis knockdown rescue, single lab","pmids":["38956029"],"is_preprint":false},{"year":2024,"finding":"WBP2 overexpression activates AMOTL2 and nuclear phosphorylated c-JUN in breast cancer cells; AMOTL2 knockdown reduces drug-resistance protein expression caused by WBP2 overexpression, placing AMOTL2 as a mediator in the ITCH/WBP2/AMOTL2/c-JUN chemoresistance axis.","method":"RNA sequencing, siRNA knockdown epistasis, immunoblotting for phospho-c-JUN, in vivo xenograft with proteasome inhibitor (MG132)","journal":"Biochemical pharmacology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — epistasis by knockdown with functional phenotype, no direct binding assay for AMOTL2/c-JUN reported, single lab","pmids":["39709035"],"is_preprint":false},{"year":2025,"finding":"AMOTL2 is identified as a direct cellular target of celastrol by activity-based protein profiling (ABPP); celastrol-AMOTL2 binding activates the Hippo pathway, promoting YAP1 phosphorylation and degradation; AMOTL2 knockdown attenuates celastrol-induced cardiomyocyte apoptosis by enhancing YAP1 expression and mitochondrial biogenesis.","method":"Activity-based protein profiling (ABPP) for direct target identification, shRNA knockdown of AMOTL2, YAP1 phosphorylation/expression assays, mitochondrial biogenesis assays, in vivo mouse cardiotoxicity model","journal":"Chemico-biological interactions","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ABPP for direct target ID plus functional KD rescue, single lab, multiple readouts","pmids":["41412440"],"is_preprint":false},{"year":2025,"finding":"AMOTL2 inhibits Zika virus replication by promoting the host type I interferon response; AMOTL2 modulates STAT1 levels and activation in response to type I IFN, promoting downstream expression of interferon-stimulated genes.","method":"CRISPR knockout screen for ZIKV host factors, AMOTL2 KO validation, STAT1 activation assays (phospho-STAT1, STAT1 protein levels), ISG expression assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genome-wide CRISPR screen with mechanistic follow-up via STAT1 assays, single lab, new function for AMOTL2","pmids":["40892926"],"is_preprint":false},{"year":2025,"finding":"AmotL2 maintains junctional architecture, actomyosin tension, and appropriate cell packing in endothelial cells; loss of AmotL2 disrupts tension homeostasis, increases tissue compaction, and prevents vascular branch regression (pruning) despite normal perfusion in zebrafish; Yap1 plays an opposing stabilizing role, and AmotL2 and Yap1 together constitute a mechanosensitive balancing module for flow-guided vascular remodeling.","method":"Zebrafish live imaging, quantitative vascular topology analysis, amotL2 and yap1 mutant/knockdown genetic analysis, actomyosin tension measurements, junctional remodeling assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo live imaging with genetic loss-of-function and quantitative tension measurements in zebrafish, preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.11.19.689183"],"is_preprint":true}],"current_model":"AMOTL2 is a scaffold protein that mechanically couples cell-cell junction proteins (VE-cadherin, E-cadherin) to contractile radial actin filaments and the nuclear lamina, transmitting mechanical forces to control cell shape and polarity; it functions as a core Hippo pathway regulator by recruiting and activating LATS1/2 kinases (via WWP1-mediated mono-ubiquitination at K347/K408) to phosphorylate and repress YAP/TAZ, while its activity is itself regulated by mTORC2 phosphorylation at S760 (which blocks YAP repression), alternative splicing (CLK2-dependent), and isoform balance (p100 vs p60); additional mechanisms include sequestering β-catenin in Rab11-positive endosomes to attenuate Wnt signaling, binding PPP2R2A to prevent nuclear JUN dephosphorylation, facilitating c-Src membrane translocation for MAPK/ERK activation, and modulating STAT1-dependent interferon signaling to restrict Zika virus replication."},"narrative":{"mechanistic_narrative":"AMOTL2 is a junctional scaffold protein that mechanically couples cadherin-based cell-cell junctions to the contractile actin cytoskeleton and the nuclear lamina, and acts as a regulatory hub for the Hippo and Wnt pathways governing cell shape, polarity, and proliferation [PMID:24806444, PMID:28842668, PMID:39195920]. At adherens junctions, the p100 AMOTL2 isoform forms complexes with VE-cadherin or E-cadherin that organize radial actin filaments, transmitting extracellular mechanical force through to the nuclear membrane; this coupling requires Par3 for junctional localization and drives lumen expansion, epithelial packing geometry, blastocyst hatching, and flow-induced endothelial alignment, with its loss in endothelium producing pro-inflammatory aortic aneurysm phenotypes [PMID:24806444, PMID:28790366, PMID:28842668, PMID:39195920]. AMOTL2 represses the transcriptional co-activators YAP/TAZ: it binds TAZ through a PPXY-WW interaction to control its nuclear translocation [PMID:23911299], and WWP1-mediated mono-ubiquitination at K347/K408 enables AMOTL2 to recruit LATS2 and SAV1 to promote YAP phosphorylation and cytoplasmic sequestration under high cell density [PMID:34404733], while it also stabilizes LATS1/2 to sustain YAP repression [PMID:38956029]. This YAP-repressive activity is switched off by mTORC2 phosphorylation at S760 and by CLK2-dependent alternative splicing that yields a membrane-uncoupled isoform [PMID:25998128, PMID:38126343]. AMOTL2 additionally attenuates Wnt signaling by trapping β-catenin in Rab11-positive recycling endosomes [PMID:22362771, PMID:34036399], and a hypoxia-induced p60 isoform sequesters Crb3/Par3 polarity complexes and uncouples p100 AMOTL2 from the actin-nuclear lamina axis to promote tumor invasion [PMID:25080976, PMID:37443716]. Through c-Src binding it facilitates membrane translocation and MAPK/ERK activation [PMID:17293535, PMID:21937427], and it modulates STAT1-dependent type I interferon signaling to restrict Zika virus replication [PMID:40892926].","teleology":[{"year":2007,"claim":"Established the first cellular role for Amotl2 by linking it to active c-Src and cytoskeletal/membrane organization during morphogenetic cell movements.","evidence":"Zebrafish morpholino knockdown, mosaic transplantation, and co-IP with phospho-c-Src and endosomal markers","pmids":["17293535"],"confidence":"Medium","gaps":["Direct binding interface with c-Src not mapped","Did not connect c-Src translocation to a specific downstream signaling output"]},{"year":2011,"claim":"Defined a tyrosine-103-dependent, PDZ-independent mechanism by which Amotl2 drives c-Src-mediated MAPK/ERK activation in endothelial morphogenesis.","evidence":"HUVEC siRNA, Y103 site-directed mutagenesis, ERK assays, and zebrafish vessel-growth readouts","pmids":["21937427"],"confidence":"High","gaps":["How Y103 phosphorylation is regulated upstream unknown","Relationship of ERK arm to junctional/Hippo functions not integrated"]},{"year":2012,"claim":"Showed Amotl2 restrains Wnt signaling by physically trapping β-catenin in recycling endosomes rather than acting at the destruction complex.","evidence":"Zebrafish epistasis with axin1 mutant, co-IP, and Rab11 endosome co-localization","pmids":["22362771"],"confidence":"High","gaps":["Determinants of β-catenin capture vs release not defined","Whether endosomal sequestration is regulated by signaling unknown"]},{"year":2013,"claim":"Identified AMOTL2 as a direct, Hippo-independent regulator of TAZ nuclear translocation via a PPXY-WW interaction, foreshadowing its central role over YAP/TAZ.","evidence":"Co-IP with domain mutagenesis (PPXY), luciferase reporter, and overexpression in H441 lung cells","pmids":["23911299"],"confidence":"Medium","gaps":["Single cell type","Did not reconcile Hippo-independent claim with later LATS-dependent mechanism"]},{"year":2013,"claim":"Placed Amotl2 at specialized actin-based adhesion structures (synaptic podosomes, invadopodia) through interaction with the scaffold LL5β.","evidence":"Affinity purification/MS, co-IP, and siRNA knockdown with morphometric analysis in myotubes and fibroblasts","pmids":["23525008"],"confidence":"Medium","gaps":["Mechanism linking LL5β binding to podosome size unknown","Not connected to cadherin/Hippo functions"]},{"year":2014,"claim":"Demonstrated that AmotL2 mechanically couples VE-cadherin junctions to contractile actin to generate the tensile forces required for vascular lumen expansion.","evidence":"Zebrafish/mouse inactivation, endothelial co-IP with VE-cadherin complex, and traction force microscopy","pmids":["24806444"],"confidence":"High","gaps":["Direct actin-binding interface not resolved","How force is sensed/transduced not yet defined"]},{"year":2014,"claim":"Revealed isoform-specific control whereby a hypoxia-induced p60 AmotL2 sequesters Crb3/Par3 polarity complexes to disrupt apical-basal polarity and drive invasion.","evidence":"c-Fos promoter analysis, co-IP with polarity complexes, vesicle trafficking assays, and xenograft","pmids":["25080976"],"confidence":"High","gaps":["Isoform-specific structural basis for polarity-complex retention unknown","p60 vs p100 abundance control incompletely defined"]},{"year":2015,"claim":"Identified mTORC2-dependent S760 phosphorylation as an off-switch for AMOTL2's YAP-repressive function, linking growth signaling to Hippo output.","evidence":"Mouse glioma gene-trap, S760A/S760E mutagenesis, co-IP with YAP, and xenograft tumor assays","pmids":["25998128"],"confidence":"High","gaps":["How S760 phosphorylation alters AMOTL2-YAP binding structurally unknown","Other mTORC2 substrates in the axis not addressed"]},{"year":2017,"claim":"Established Par3 as the localization determinant placing AmotL2 at junctions to build the cadherin-actin axis controlling tissue geometry.","evidence":"Zebrafish inactivation, cell-culture siRNA, immunofluorescence, and co-IP","pmids":["28790366"],"confidence":"Medium","gaps":["Direct vs indirect Par3-AmotL2 interaction not distinguished","Single lab"]},{"year":2017,"claim":"Showed the p100 isoform forms an E-cadherin/radial-actin complex that generates the contractile tension required for blastocyst hatching and epithelial packing.","evidence":"Co-IP, zebrafish/mouse inactivation, and blebbistatin phenocopy of myosin-II loss","pmids":["28842668"],"confidence":"High","gaps":["Stoichiometry of E-cadherin/AmotL2/actin complex unresolved","Quantitative force contribution not measured"]},{"year":2020,"claim":"Uncovered a junction-independent role for AMOTL2 in sequestering the PP2A subunit PPP2R2A in the cytoplasm to sustain phospho-JUN and AP-1 proliferation signaling.","evidence":"CRISPR loss-of-function screen, MS, co-IP/GST pull-down, fractionation, and phospho-JUN immunoblot in NSCLC","pmids":["32950569"],"confidence":"High","gaps":["Whether PPP2R2A binding competes with YAP/cadherin functions unknown","Regulation of this interaction not defined"]},{"year":2021,"claim":"Defined WWP1-mediated mono-ubiquitination at K347/K408 as the molecular trigger that enables AMOTL2 to recruit LATS2/SAV1 and drive YAP phosphorylation at high cell density.","evidence":"In vitro ubiquitination, K347/K408 mutagenesis, co-IP with LATS2/SAV1, and contact-inhibition assays","pmids":["34404733"],"confidence":"High","gaps":["Deubiquitinase counteracting WWP1 not identified","How Crumbs coupling is regulated mechanistically unresolved"]},{"year":2021,"claim":"Extended the β-catenin sequestration mechanism to glioma, showing AMOTL2 loss enhances Wnt-driven proliferation and invasion.","evidence":"Co-IP, immunofluorescence, and siRNA knockdown in glioma cell lines","pmids":["34036399"],"confidence":"Medium","gaps":["Single lab, no in vivo validation","Did not test endosomal trapping specifically"]},{"year":2021,"claim":"Placed AMOTL2 downstream of MAGI1 loss as an upstream driver of a ROCK/p38 stress pathway in breast cancer.","evidence":"Double-knockdown epistasis, ROCK/p38 inhibitor rescue, and cell-stiffness/tumorigenicity assays","pmids":["33707576"],"confidence":"Medium","gaps":["Direct molecular link from AMOTL2 to ROCK/p38 not shown","In vitro only"]},{"year":2021,"claim":"Confirmed AMOTL2 restrains TGF-β1-induced YAP1 activation in airway smooth muscle, linking it to proliferation and ECM deposition.","evidence":"Overexpression/knockdown in ASM cells with YAP1 nuclear-localization and constitutively-active YAP1 rescue","pmids":["34323359"],"confidence":"Medium","gaps":["No direct AMOTL2-YAP1 binding assay reported","Single lab"]},{"year":2022,"claim":"Showed AMOTL2 is transcriptionally repressed by MEF2D, derepressing YAP signaling to promote hepatocellular carcinoma.","evidence":"Dual luciferase reporter and overexpression in hepatoma cells","pmids":["35698637"],"confidence":"Low","gaps":["Limited mechanistic depth; promoter binding inferred","No in vivo confirmation"]},{"year":2023,"claim":"Identified CLK2-dependent alternative splicing as a regulator that generates a membrane-uncoupled AMOTL2 isoform, switching off YAP repression.","evidence":"Chemical screen, RT-PCR splicing analysis, and YAP phosphorylation/localization assays with CLK2 inhibitor","pmids":["38126343"],"confidence":"Medium","gaps":["Spliced isoform binding partners not reconstituted","Physiological context of CLK2 control unclear"]},{"year":2023,"claim":"Revealed the mechanism of p60-driven invasion: p60 binds p100 AMOTL2 to uncouple E-cadherin-radial actin from the nuclear lamina, softening the nucleus for ameboid migration.","evidence":"Inter-isoform co-IP, atomic force microscopy of nuclear stiffness, and micropore invasion assays","pmids":["37443716"],"confidence":"Medium","gaps":["Direct nuclear-lamina binding partner not identified","Single lab"]},{"year":2023,"claim":"Demonstrated in vivo that AmotL2 transmits flow-derived mechanical signals from VE-cadherin junctions through actin to the nuclear lamina, with endothelial loss causing aortic aneurysms.","evidence":"Endothelial-specific mouse knockout, tripartite co-IP, nuclear-lamina connectivity assays, and transcriptomics","pmids":["39195920"],"confidence":"High","gaps":["Identity of the nuclear-lamina anchoring partner unresolved","Mechanotransduction signaling steps to inflammation not fully mapped"]},{"year":2024,"claim":"Identified ARNTL2 as a transcriptional repressor of AMOTL2 that, by reducing LATS1/2 recruitment and stabilization, activates YAP to promote nasopharyngeal carcinoma invasion.","evidence":"ChIP, knockdown epistasis with rescue, LATS1/2 co-IP, and xenograft","pmids":["38956029"],"confidence":"Medium","gaps":["Direct LATS1/2 stabilization mechanism vs recruitment not separated","Single lab"]},{"year":2024,"claim":"Positioned AMOTL2 as a mediator in a WBP2/c-JUN chemoresistance axis in breast cancer.","evidence":"RNA-seq, knockdown epistasis, phospho-c-JUN immunoblot, and xenograft with MG132","pmids":["39709035"],"confidence":"Low","gaps":["No direct AMOTL2/c-JUN binding assay","Mechanism of drug-resistance protein induction unclear"]},{"year":2025,"claim":"Identified AMOTL2 as a direct binding target of the natural product celastrol, linking pharmacological engagement to Hippo activation and cardiomyocyte apoptosis.","evidence":"Activity-based protein profiling, shRNA knockdown rescue, YAP1 phosphorylation and mitochondrial biogenesis assays, and mouse cardiotoxicity model","pmids":["41412440"],"confidence":"Medium","gaps":["Celastrol binding site on AMOTL2 not mapped","Whether binding mimics a physiological regulatory event unknown"]},{"year":2025,"claim":"Uncovered an antiviral role in which AMOTL2 promotes the type I interferon response by modulating STAT1 to restrict Zika virus replication.","evidence":"Genome-wide CRISPR knockout screen for ZIKV host factors with STAT1 activation and ISG expression follow-up","pmids":["40892926"],"confidence":"Medium","gaps":["Direct molecular interaction with STAT1 not demonstrated","Relation to junctional/Hippo roles unknown"]},{"year":2025,"claim":"Showed AmotL2 and Yap1 form an opposing mechanosensitive module balancing actomyosin tension to enable flow-guided vascular pruning.","evidence":"Zebrafish live imaging, amotL2/yap1 genetic analysis, and actomyosin tension measurements (preprint)","pmids":["bio_10.1101_2025.11.19.689183"],"confidence":"Medium","gaps":["Preprint not yet peer-reviewed","Molecular basis of the AmotL2-Yap1 balance during pruning not resolved"]},{"year":null,"claim":"The structural basis by which AMOTL2 physically bridges cadherin, actin, and the nuclear lamina, and how its multiple regulatory inputs (ubiquitination, phosphorylation, splicing, isoform balance, transcriptional repression) are integrated within a single cell, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of AMOTL2 complexes","Nuclear-lamina anchoring partner unidentified","Cross-talk between Hippo, Wnt, MAPK, and interferon roles not integrated"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,5,9,11,18]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[5,9,18]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[7,11,19,14]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[2,6,10]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,5,8,18]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3,10]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[2,6]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[5,9,18]},{"term_id":"GO:0005635","term_label":"nuclear envelope","supporting_discovery_ids":[17,18]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,7,11,19]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[5,8,9,18]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[22]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[6,10,17]}],"complexes":["VE-cadherin adherens junction complex","E-cadherin/p100-AMOTL2/radial actin complex","LATS2/SAV1 Hippo complex"],"partners":["CDH5","CDH1","TAZ/WWTR1","YAP1","LATS2","CTNNB1","PPP2R2A","PARD3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y2J4","full_name":"Angiomotin-like protein 2","aliases":["Leman coiled-coil protein","LCCP"],"length_aa":779,"mass_kda":85.8,"function":"Regulates the translocation of phosphorylated SRC to peripheral cell-matrix adhesion sites. Required for proper architecture of actin filaments. Plays a role in coupling actin fibers to cell junctions in endothelial cells and is therefore required for correct endothelial cell morphology via facilitating transcellular transmission of mechanical force resulting in endothelial cell elongation (By similarity). Required for the anchoring of radial actin fibers to CDH1 junction complexes at the cell membrane which facilitates organization of radial actin fiber structure and cellular response to contractile forces (PubMed:28842668). This contributes to maintenance of cell area, size, shape, epithelial sheet organization and trophectoderm cell properties that facilitate blastocyst zona hatching (PubMed:28842668). Inhibits the Wnt/beta-catenin signaling pathway, probably by recruiting CTNNB1 to recycling endosomes and hence preventing its translocation to the nucleus. Participates in angiogenesis. Activates the Hippo signaling pathway in response to cell contact inhibition via interaction with and ubiquitination by Crumbs complex-bound WWP1 (PubMed:34404733). Ubiquitinated AMOTL2 then interacts with LATS2 which in turn phosphorylates YAP1, excluding it from the nucleus and localizing it to the cytoplasm and tight junctions, therefore ultimately repressing YAP1-driven transcription of target genes (PubMed:17293535, PubMed:21205866, PubMed:26598551). Acts to inhibit WWTR1/TAZ transcriptional coactivator activity via sequestering WWTR1/TAZ in the cytoplasm and at tight junctions (PubMed:23911299). Regulates the size and protein composition of the podosome cortex and core at myofibril neuromuscular junctions (PubMed:23525008). Selectively promotes FGF-induced MAPK activation through SRC (PubMed:17293535). May play a role in the polarity, proliferation and migration of endothelial cells","subcellular_location":"Recycling endosome; Cytoplasm; Cell projection, podosome; Cell junction","url":"https://www.uniprot.org/uniprotkb/Q9Y2J4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMOTL2","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"DYNLL1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/AMOTL2","total_profiled":1310},"omim":[{"mim_id":"614658","title":"ANGIOMOTIN-LIKE 2; AMOTL2","url":"https://www.omim.org/entry/614658"},{"mim_id":"614657","title":"ANGIOMOTIN-LIKE 1; AMOTL1","url":"https://www.omim.org/entry/614657"},{"mim_id":"300410","title":"ANGIOMOTIN; AMOT","url":"https://www.omim.org/entry/300410"},{"mim_id":"222900","title":"SUCRASE-ISOMALTASE DEFICIENCY, CONGENITAL; CSID","url":"https://www.omim.org/entry/222900"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cell Junctions","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/AMOTL2"},"hgnc":{"alias_symbol":["LCCP"],"prev_symbol":[]},"alphafold":{"accession":"Q9Y2J4","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y2J4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y2J4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y2J4-F1-predicted_aligned_error_v6.png","plddt_mean":65.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AMOTL2","jax_strain_url":"https://www.jax.org/strain/search?query=AMOTL2"},"sequence":{"accession":"Q9Y2J4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y2J4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y2J4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y2J4"}},"corpus_meta":[{"pmid":"32942233","id":"PMC_32942233","title":"Hypoxic Tumor-Derived Exosomal Long Noncoding RNA UCA1 Promotes Angiogenesis via miR-96-5p/AMOTL2 in Pancreatic Cancer.","date":"2020","source":"Molecular therapy. Nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/32942233","citation_count":153,"is_preprint":false},{"pmid":"25998128","id":"PMC_25998128","title":"Phosphorylation of the Hippo Pathway Component AMOTL2 by the mTORC2 Kinase Promotes YAP Signaling, Resulting in Enhanced Glioblastoma Growth and Invasiveness.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25998128","citation_count":81,"is_preprint":false},{"pmid":"21937427","id":"PMC_21937427","title":"Angiomotin-like2 gene (amotl2) is required for migration and proliferation of endothelial cells during angiogenesis.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21937427","citation_count":64,"is_preprint":false},{"pmid":"24806444","id":"PMC_24806444","title":"AmotL2 links VE-cadherin to contractile actin fibres necessary for aortic lumen expansion.","date":"2014","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/24806444","citation_count":60,"is_preprint":false},{"pmid":"25080976","id":"PMC_25080976","title":"AmotL2 disrupts apical-basal cell polarity and promotes tumour invasion.","date":"2014","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/25080976","citation_count":53,"is_preprint":false},{"pmid":"17293535","id":"PMC_17293535","title":"Amotl2 is essential for cell movements in zebrafish embryo and regulates c-Src translocation.","date":"2007","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/17293535","citation_count":49,"is_preprint":false},{"pmid":"22362771","id":"PMC_22362771","title":"The Amotl2 gene inhibits Wnt/β-catenin signaling and regulates embryonic development in zebrafish.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22362771","citation_count":47,"is_preprint":false},{"pmid":"23525008","id":"PMC_23525008","title":"Amotl2 interacts with LL5β, localizes to podosomes and regulates postsynaptic differentiation in muscle.","date":"2013","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/23525008","citation_count":44,"is_preprint":false},{"pmid":"28842668","id":"PMC_28842668","title":"The E-cadherin/AmotL2 complex organizes actin filaments required for epithelial hexagonal packing and blastocyst hatching.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28842668","citation_count":26,"is_preprint":false},{"pmid":"32950569","id":"PMC_32950569","title":"AMOTL2 inhibits JUN Thr239 dephosphorylation by binding PPP2R2A to suppress the proliferation in non-small cell lung cancer cells.","date":"2020","source":"Biochimica et biophysica acta. Molecular cell research","url":"https://pubmed.ncbi.nlm.nih.gov/32950569","citation_count":19,"is_preprint":false},{"pmid":"39195920","id":"PMC_39195920","title":"The VE-cadherin/AmotL2 mechanosensory pathway suppresses aortic inflammation and the formation of abdominal aortic aneurysms.","date":"2023","source":"Nature cardiovascular research","url":"https://pubmed.ncbi.nlm.nih.gov/39195920","citation_count":14,"is_preprint":false},{"pmid":"38956029","id":"PMC_38956029","title":"The circadian gene ARNTL2 promotes nasopharyngeal carcinoma invasiveness and metastasis through suppressing AMOTL2-LATS-YAP pathway.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/38956029","citation_count":12,"is_preprint":false},{"pmid":"34165350","id":"PMC_34165350","title":"AmotL2, IQGAP1, and FKBP51 Scaffold Proteins in Glioblastoma Stem Cell Niches.","date":"2021","source":"The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society","url":"https://pubmed.ncbi.nlm.nih.gov/34165350","citation_count":11,"is_preprint":false},{"pmid":"33707576","id":"PMC_33707576","title":"MAGI1 inhibits the AMOTL2/p38 stress pathway and prevents luminal breast tumorigenesis.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33707576","citation_count":11,"is_preprint":false},{"pmid":"28790366","id":"PMC_28790366","title":"AmotL2 integrates polarity and junctional cues to modulate cell shape.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28790366","citation_count":10,"is_preprint":false},{"pmid":"34036399","id":"PMC_34036399","title":"AMOTL2‑knockdown promotes the proliferation, migration and invasion of glioma by regulating β‑catenin nuclear localization.","date":"2021","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/34036399","citation_count":10,"is_preprint":false},{"pmid":"23911299","id":"PMC_23911299","title":"AMOTL2 interaction with TAZ causes the inhibition of surfactant proteins expression in lung cells.","date":"2013","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/23911299","citation_count":10,"is_preprint":false},{"pmid":"35563891","id":"PMC_35563891","title":"FKBP51, AmotL2 and IQGAP1 Involvement in Cilastatin Prevention of Cisplatin-Induced Tubular Nephrotoxicity in Rats.","date":"2022","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/35563891","citation_count":8,"is_preprint":false},{"pmid":"34323359","id":"PMC_34323359","title":"AMOTL2 restrains transforming growth factor-β1-induced proliferation and extracellular matrix deposition of airway smooth muscle cells via the down-regulation of YAP1 activation.","date":"2021","source":"Environmental toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/34323359","citation_count":8,"is_preprint":false},{"pmid":"39709035","id":"PMC_39709035","title":"E3 ubiquitin ligase ITCH-mediated proteasomal degradation of WBP2 sensitizes breast cancer cells to chemotherapy through restraining AMOTL2/c-JUN axis.","date":"2024","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39709035","citation_count":8,"is_preprint":false},{"pmid":"35698637","id":"PMC_35698637","title":"Myocyte enhancer factor 2D promotes hepatocellular carcinoma through AMOTL2/YAP signaling that inhibited by luteolin.","date":"2022","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/35698637","citation_count":6,"is_preprint":false},{"pmid":"39467346","id":"PMC_39467346","title":"LCCP exposure leads to skin cell senescence damage by triggering oxidative stress mediated by mitochondrial Ca2+ overload.","date":"2024","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39467346","citation_count":5,"is_preprint":false},{"pmid":"34404733","id":"PMC_34404733","title":"AMOTL2 mono-ubiquitination by WWP1 promotes contact inhibition by facilitating LATS activation.","date":"2021","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/34404733","citation_count":4,"is_preprint":false},{"pmid":"39335466","id":"PMC_39335466","title":"Changes in AmotL2 Expression in Cells of the Human Enteral Nervous System in Oxaliplatin-Induced Enteric Neuropathy.","date":"2024","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/39335466","citation_count":4,"is_preprint":false},{"pmid":"38126343","id":"PMC_38126343","title":"Pharmacological inhibition of CLK2 activates YAP by promoting alternative splicing of AMOTL2.","date":"2023","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/38126343","citation_count":3,"is_preprint":false},{"pmid":"40154222","id":"PMC_40154222","title":"Bone marrow mesenchymal stem cells senescence induced by LCCP through activation of cGAS-STING-mediated inflammation.","date":"2025","source":"Ecotoxicology and environmental safety","url":"https://pubmed.ncbi.nlm.nih.gov/40154222","citation_count":2,"is_preprint":false},{"pmid":"37443716","id":"PMC_37443716","title":"Modulation of E-Cadherin Function through the AmotL2 Isoforms Promotes Ameboid Cell Invasion.","date":"2023","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/37443716","citation_count":2,"is_preprint":false},{"pmid":"41412440","id":"PMC_41412440","title":"Celastrol induces cardiotoxicity by directly targeting AMOTL2 and inhibiting YAP1/PGC-1α/TFAM-dependent mitochondrial biogenesis.","date":"2025","source":"Chemico-biological interactions","url":"https://pubmed.ncbi.nlm.nih.gov/41412440","citation_count":1,"is_preprint":false},{"pmid":"40892926","id":"PMC_40892926","title":"Identification of AMOTL2 as an antiviral factor that enhances the human type I interferon response against Zika virus.","date":"2025","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/40892926","citation_count":1,"is_preprint":false},{"pmid":"37131806","id":"PMC_37131806","title":"Pharmacological inhibition of CLK2 activates YAP by promoting alternative splicing of AMOTL2.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/37131806","citation_count":0,"is_preprint":false},{"pmid":"40569478","id":"PMC_40569478","title":"Circ_0001461 Regulates Colorectal Cancer Growth, Movement and Immune Escape Through miR-532-3p/AMOTL2.","date":"2025","source":"Biochemical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/40569478","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.11.19.689183","title":"An AmotL2–Yap1 Module Integrates Flow and Junctional Mechanics to Specify Vascular Pruning Hotspots","date":"2025-11-19","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.19.689183","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.11.13.24317257","title":"Single cell variant to enhancer to gene map for coronary artery disease","date":"2024-11-13","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.13.24317257","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":16891,"output_tokens":6361,"usd":0.073044,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15153,"output_tokens":6780,"usd":0.122633,"stage2_stop_reason":"end_turn"},"total_usd":0.195677,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"Amotl2 interacts preferentially with phosphorylated c-Src and facilitates its outward translocation to the membrane, regulating membrane architecture and F-actin organization; knockdown of amotl2 in zebrafish delays epiboly and impairs convergence/extension cell movements, and amotl2-deficient cells fail to migrate properly with loss of membrane protrusions.\",\n      \"method\": \"Zebrafish morpholino knockdown, mosaic transplantation, co-immunoprecipitation, co-localization with endosomal markers (RhoB, EEA1)\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP and in vivo loss-of-function in zebrafish with defined cellular phenotypes, single lab\",\n      \"pmids\": [\"17293535\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Amotl2 promotes MAPK/ERK activation via c-Src in endothelial cells, dependent on phosphorylation of tyrosine residue at position 103 but independent of the C-terminal PDZ-binding domain; knockdown impairs endothelial cell proliferation, migration, polarity, and tube formation in vitro, and intersegmental vessel growth in zebrafish.\",\n      \"method\": \"Zebrafish transgenic line knockdown, HUVEC siRNA knockdown, site-directed mutagenesis (Y103), ERK activity assays, cell polarity and tube formation assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — mutagenesis combined with in vitro biochemical assays and in vivo zebrafish validation, two orthogonal model systems\",\n      \"pmids\": [\"21937427\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Amotl2 attenuates Wnt/β-catenin signaling by associating with and trapping β-catenin in Rab11-positive recycling endosomes, reducing the amount of β-catenin available in the cytosol and nucleus; knockdown in zebrafish causes embryonic dorsalization rescued by co-knockdown of β-catenin2.\",\n      \"method\": \"Zebrafish morpholino knockdown, genetic epistasis with axin1 mutant (masterblind), co-immunoprecipitation, immunofluorescence with Rab11 co-localization in mammalian cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, co-localization with Rab11 endosomes, in vivo epistasis in zebrafish, replicated across cell and animal models\",\n      \"pmids\": [\"22362771\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AMOTL2 co-immunoprecipitates with TAZ via the WW domain of TAZ and the PPXY motif in the N-terminus of AMOTL2; AMOTL2 co-localizes with TAZ in the cytoplasm and regulates TAZ cytoplasm-to-nucleus translocation through direct protein-protein interaction, inhibiting TAZ-dependent transcription of surfactant genes in lung cells in a Hippo-independent manner.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence co-localization, luciferase reporter assay, domain mutagenesis (PPXY motif), overexpression in H441 lung cells\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain mapping and reporter assay, single lab, single cell type\",\n      \"pmids\": [\"23911299\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Amotl2 interacts with the scaffold protein LL5β at synaptic podosomes in myotubes and neuromuscular junctions in vivo; depletion of Amotl2 in myotubes increases size of synaptic podosomes and alters postsynaptic topology, and depletion in fibroblasts disrupts invadopodia.\",\n      \"method\": \"Affinity purification/mass spectrometry (LL5β-associated proteins), co-immunoprecipitation, immunofluorescence in vivo and in vitro, siRNA knockdown with morphometric analysis\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-based identification followed by co-IP and loss-of-function with defined phenotype, single lab\",\n      \"pmids\": [\"23525008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"AmotL2 associates with the VE-cadherin adherens junction complex and couples it to contractile actin fibres; inactivation of amotL2 in zebrafish, mouse, and endothelial cell culture dissociates VE-cadherin from cytoskeletal tensile forces, impairing aortic vessel lumen expansion.\",\n      \"method\": \"Gene inactivation in zebrafish and mouse, endothelial cell culture knockdown, co-immunoprecipitation with VE-cadherin complex, traction force microscopy, live imaging\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal model systems (zebrafish, mouse, cell culture), co-IP with junctional complex, functional mechanical readouts across independent labs\",\n      \"pmids\": [\"24806444\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Hypoxic stress induces c-Fos-dependent expression of a p60 AmotL2 isoform; p60 AmotL2 interacts with the Crb3 and Par3 polarity complexes, retaining them in large vesicles and preventing them from reaching the apical membrane, causing loss of apical-basal polarity and promoting tumor invasion.\",\n      \"method\": \"Immunoprecipitation, immunofluorescence, siRNA knockdown in vitro, mouse xenograft in vivo, c-Fos promoter analysis, vesicle trafficking assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP with polarity complexes, mechanistic promoter analysis, in vitro and in vivo validation with defined cellular phenotype\",\n      \"pmids\": [\"25080976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"mTORC2 phosphorylates AMOTL2 at serine 760; phosphomimetic S760E mutation blocks AMOTL2's ability to bind and repress YAP, increasing YAP target gene expression, foci formation, and metastatic properties, whereas non-phosphorylatable S760A mutant retains YAP repression activity in glioblastoma cells and xenografts.\",\n      \"method\": \"Gene-trap in mouse glioma model, phosphosite mutagenesis (S760A/S760E), co-immunoprecipitation with YAP, YAP transcriptional reporter assays, foci formation assay, xenograft tumor growth\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — site-directed mutagenesis with biochemical and in vivo validation, multiple orthogonal phenotypic readouts, in vitro and in vivo\",\n      \"pmids\": [\"25998128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Par3 is essential for localization of AmotL2 to cellular junctions, where it associates with VE/E-cadherin to organize radial actin filaments; loss of this Par3-AmotL2-cadherin-actin axis impairs aortic lumen expansion and epithelial hexagonal packing.\",\n      \"method\": \"Gene inactivation in zebrafish, siRNA knockdown in cell culture, immunofluorescence, co-immunoprecipitation\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP and in vivo genetic epistasis in zebrafish, single lab, two orthogonal systems\",\n      \"pmids\": [\"28790366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"p100 AmotL2 forms a complex with E-cadherin that associates with radial actin filaments connecting cells across multiple layers; genetic inactivation of amotL2 leads to loss of contractile actin filaments and perturbed epithelial packing geometry; amotL2 is required for blastocyst hatching in mouse and zebrafish via tension generation, phenocopied by myosin II inhibitor blebbistatin.\",\n      \"method\": \"Co-immunoprecipitation, zebrafish and mouse genetic inactivation, immunofluorescence, blebbistatin pharmacological inhibition, blastocyst hatching assay\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP, multiple genetic model systems (zebrafish, mouse), pharmacological inhibition corroborating mechanism, defined developmental phenotype\",\n      \"pmids\": [\"28842668\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AMOTL2 is a binding partner of PPP2R2A (a PP2A regulatory subunit) in NSCLC cells; AMOTL2 binds PPP2R2A in the cytoplasm, reducing its nuclear localization and thereby preventing PPP2R2A-mediated dephosphorylation of JUN at Thr239, which suppresses AP-1-driven cell proliferation.\",\n      \"method\": \"CRISPR/Cas9 loss-of-function screen, mass spectrometry, co-immunoprecipitation, GST pull-down, immunofluorescence, subcellular fractionation, phospho-JUN immunoblotting\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal binding assays (MS, co-IP, GST pull-down), functional mechanistic follow-up with subcellular localization and phosphorylation readout\",\n      \"pmids\": [\"32950569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"E3 ubiquitin ligase WWP1 mono-ubiquitinates AMOTL2 at K347 and K408; mono-ubiquitinated AMOTL2 interacts with LATS2 and facilitates recruitment of SAV1, promoting YAP phosphorylation and cytoplasmic sequestration/degradation; this process is coupled to Crumbs polarity complex at cell junctions under high cell density conditions.\",\n      \"method\": \"In vitro ubiquitination assay, ubiquitination site mutagenesis (K347/K408), co-immunoprecipitation with LATS2/SAV1, YAP phosphorylation immunoblotting, cell density contact inhibition assays, immunofluorescence\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — in vitro ubiquitination with site mutagenesis plus co-IP and functional YAP phosphorylation readout, multiple orthogonal methods\",\n      \"pmids\": [\"34404733\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AMOTL2 directly binds β-catenin and regulates its nuclear translocation in glioma cells; knockdown of AMOTL2 promotes β-catenin nuclear localization and downstream Wnt target gene expression, enhancing glioma proliferation, migration, and invasion.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, immunoblotting, siRNA knockdown in glioma cell lines\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — co-IP and immunofluorescence with functional loss-of-function assays, single lab, consistent with prior Wnt pathway findings\",\n      \"pmids\": [\"34036399\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Loss of MAGI1 causes accumulation of E-cadherin and AMOTL2 and increased ROCK and p38 SAPK activities; rescue experiments show that AMOTL2 depletion or p38 inhibition reverses the increased tumorigenicity of MAGI1-deficient cells, placing AMOTL2 upstream of a ROCK/p38 stress pathway in luminal breast cancer.\",\n      \"method\": \"Genetic epistasis (double knockdown), ROCK/p38 inhibitor pharmacology, cell stiffness measurement, tumorigenicity assays in vitro\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via double-depletion and pharmacological rescue, single lab, functional phenotype defined\",\n      \"pmids\": [\"33707576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AMOTL2 restrains YAP1 activation in airway smooth muscle cells; overexpression of AMOTL2 suppresses TGF-β1-induced YAP1 nuclear translocation, and reactivation of YAP1 reverses AMOTL2-mediated suppression of proliferation and ECM deposition.\",\n      \"method\": \"Overexpression and siRNA knockdown in ASM cells, YAP1 nuclear localization assay, proliferation and ECM deposition functional assays, rescue with constitutively active YAP1\",\n      \"journal\": \"Environmental toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, overexpression/KD with functional readout and YAP1 rescue, no direct binding assay reported in abstract\",\n      \"pmids\": [\"34323359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MEF2D binds the MEF2 cis-acting element in the upstream promoter region of AMOTL2, inhibiting its transcriptional expression, thereby activating YAP signaling and promoting HCC cell migration and proliferation.\",\n      \"method\": \"Dual luciferase reporter assay, ChIP (implied by promoter binding), overexpression experiments in hepatoma cells\",\n      \"journal\": \"International journal of clinical and experimental pathology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — reporter assay for promoter binding, single lab, limited mechanistic depth described in abstract\",\n      \"pmids\": [\"35698637\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CLK2 inhibition promotes alternative splicing of AMOTL2 producing an exon-skipped isoform that can no longer associate with membrane-bound proteins, resulting in decreased YAP phosphorylation and decreased membrane localization of YAP, thereby activating YAP-driven transcription.\",\n      \"method\": \"High-throughput chemical screen, RT-PCR for alternative splicing, YAP phosphorylation and localization assays, CLK2 inhibitor (SM04690) pharmacology\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chemical-genetic approach with splicing and functional YAP readouts, single lab, mechanism defined but no direct binding reconstitution\",\n      \"pmids\": [\"38126343\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"p60 AmotL2 (expressed in invading tumor cells) binds to the p100 AmotL2 isoform and uncouples the mechanical constraint of radial actin filaments from E-cadherin; the E-cadherin/p100AmotL2 complex is directly connected to the nuclear membrane, and p60AmotL2 expression inactivates this connection, altering nuclear lamina properties and potentiating ameboid invasion through extracellular matrix micropores.\",\n      \"method\": \"Co-immunoprecipitation between isoforms, nuclear lamina connectivity assays, atomic force microscopy, micropore invasion assays, overexpression/knockdown in tumor cell lines\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP between isoforms plus functional invasion assay and biophysical nuclear stiffness measurement, single lab\",\n      \"pmids\": [\"37443716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AmotL2 connects junctional VE-cadherin and actin filaments to the nuclear lamina in endothelial cells; AmotL2 is essential for radial actin filament formation and endothelial cell alignment in response to blood flow; loss of endothelial AmotL2 in mice causes a pro-inflammatory response and abdominal aortic aneurysms. Molecular analysis showed VE-cadherin binds AmotL2 and actin, transmitting extracellular mechanical signals to the nuclear membrane.\",\n      \"method\": \"Endothelial-specific mouse knockout, co-immunoprecipitation (VE-cadherin/AmotL2/actin), immunofluorescence for nuclear lamina connectivity, transcriptome analysis, live imaging\",\n      \"journal\": \"Nature cardiovascular research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — endothelial-specific KO mouse with defined vascular disease phenotype, co-IP demonstrating tripartite complex, nuclear lamina connectivity assay, transcriptome validation\",\n      \"pmids\": [\"39195920\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ARNTL2 negatively regulates AMOTL2 transcription by directly binding to the AMOTL2 promoter; reduced AMOTL2 decreases its recruitment and stabilization of LATS1/2 kinases, reducing LATS-dependent YAP phosphorylation and promoting YAP nuclear translocation and NPC invasion; inhibition of AMOTL2 counteracted the effect of ARNTL2 knockdown.\",\n      \"method\": \"ChIP (ARNTL2 binding to AMOTL2 promoter), siRNA/shRNA knockdown epistasis, LATS1/2 co-immunoprecipitation with AMOTL2, YAP phosphorylation and localization assays, xenograft mouse model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP for promoter binding, co-IP for LATS interaction, epistasis knockdown rescue, single lab\",\n      \"pmids\": [\"38956029\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"WBP2 overexpression activates AMOTL2 and nuclear phosphorylated c-JUN in breast cancer cells; AMOTL2 knockdown reduces drug-resistance protein expression caused by WBP2 overexpression, placing AMOTL2 as a mediator in the ITCH/WBP2/AMOTL2/c-JUN chemoresistance axis.\",\n      \"method\": \"RNA sequencing, siRNA knockdown epistasis, immunoblotting for phospho-c-JUN, in vivo xenograft with proteasome inhibitor (MG132)\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — epistasis by knockdown with functional phenotype, no direct binding assay for AMOTL2/c-JUN reported, single lab\",\n      \"pmids\": [\"39709035\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AMOTL2 is identified as a direct cellular target of celastrol by activity-based protein profiling (ABPP); celastrol-AMOTL2 binding activates the Hippo pathway, promoting YAP1 phosphorylation and degradation; AMOTL2 knockdown attenuates celastrol-induced cardiomyocyte apoptosis by enhancing YAP1 expression and mitochondrial biogenesis.\",\n      \"method\": \"Activity-based protein profiling (ABPP) for direct target identification, shRNA knockdown of AMOTL2, YAP1 phosphorylation/expression assays, mitochondrial biogenesis assays, in vivo mouse cardiotoxicity model\",\n      \"journal\": \"Chemico-biological interactions\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ABPP for direct target ID plus functional KD rescue, single lab, multiple readouts\",\n      \"pmids\": [\"41412440\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AMOTL2 inhibits Zika virus replication by promoting the host type I interferon response; AMOTL2 modulates STAT1 levels and activation in response to type I IFN, promoting downstream expression of interferon-stimulated genes.\",\n      \"method\": \"CRISPR knockout screen for ZIKV host factors, AMOTL2 KO validation, STAT1 activation assays (phospho-STAT1, STAT1 protein levels), ISG expression assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genome-wide CRISPR screen with mechanistic follow-up via STAT1 assays, single lab, new function for AMOTL2\",\n      \"pmids\": [\"40892926\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AmotL2 maintains junctional architecture, actomyosin tension, and appropriate cell packing in endothelial cells; loss of AmotL2 disrupts tension homeostasis, increases tissue compaction, and prevents vascular branch regression (pruning) despite normal perfusion in zebrafish; Yap1 plays an opposing stabilizing role, and AmotL2 and Yap1 together constitute a mechanosensitive balancing module for flow-guided vascular remodeling.\",\n      \"method\": \"Zebrafish live imaging, quantitative vascular topology analysis, amotL2 and yap1 mutant/knockdown genetic analysis, actomyosin tension measurements, junctional remodeling assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo live imaging with genetic loss-of-function and quantitative tension measurements in zebrafish, preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.11.19.689183\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"AMOTL2 is a scaffold protein that mechanically couples cell-cell junction proteins (VE-cadherin, E-cadherin) to contractile radial actin filaments and the nuclear lamina, transmitting mechanical forces to control cell shape and polarity; it functions as a core Hippo pathway regulator by recruiting and activating LATS1/2 kinases (via WWP1-mediated mono-ubiquitination at K347/K408) to phosphorylate and repress YAP/TAZ, while its activity is itself regulated by mTORC2 phosphorylation at S760 (which blocks YAP repression), alternative splicing (CLK2-dependent), and isoform balance (p100 vs p60); additional mechanisms include sequestering β-catenin in Rab11-positive endosomes to attenuate Wnt signaling, binding PPP2R2A to prevent nuclear JUN dephosphorylation, facilitating c-Src membrane translocation for MAPK/ERK activation, and modulating STAT1-dependent interferon signaling to restrict Zika virus replication.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AMOTL2 is a junctional scaffold protein that mechanically couples cadherin-based cell-cell junctions to the contractile actin cytoskeleton and the nuclear lamina, and acts as a regulatory hub for the Hippo and Wnt pathways governing cell shape, polarity, and proliferation [#5, #9, #18]. At adherens junctions, the p100 AMOTL2 isoform forms complexes with VE-cadherin or E-cadherin that organize radial actin filaments, transmitting extracellular mechanical force through to the nuclear membrane; this coupling requires Par3 for junctional localization and drives lumen expansion, epithelial packing geometry, blastocyst hatching, and flow-induced endothelial alignment, with its loss in endothelium producing pro-inflammatory aortic aneurysm phenotypes [#5, #8, #9, #18]. AMOTL2 represses the transcriptional co-activators YAP/TAZ: it binds TAZ through a PPXY-WW interaction to control its nuclear translocation [#3], and WWP1-mediated mono-ubiquitination at K347/K408 enables AMOTL2 to recruit LATS2 and SAV1 to promote YAP phosphorylation and cytoplasmic sequestration under high cell density [#11], while it also stabilizes LATS1/2 to sustain YAP repression [#19]. This YAP-repressive activity is switched off by mTORC2 phosphorylation at S760 and by CLK2-dependent alternative splicing that yields a membrane-uncoupled isoform [#7, #16]. AMOTL2 additionally attenuates Wnt signaling by trapping \\u03b2-catenin in Rab11-positive recycling endosomes [#2, #12], and a hypoxia-induced p60 isoform sequesters Crb3/Par3 polarity complexes and uncouples p100 AMOTL2 from the actin-nuclear lamina axis to promote tumor invasion [#6, #17]. Through c-Src binding it facilitates membrane translocation and MAPK/ERK activation [#0, #1], and it modulates STAT1-dependent type I interferon signaling to restrict Zika virus replication [#22].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established the first cellular role for Amotl2 by linking it to active c-Src and cytoskeletal/membrane organization during morphogenetic cell movements.\",\n      \"evidence\": \"Zebrafish morpholino knockdown, mosaic transplantation, and co-IP with phospho-c-Src and endosomal markers\",\n      \"pmids\": [\"17293535\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding interface with c-Src not mapped\", \"Did not connect c-Src translocation to a specific downstream signaling output\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined a tyrosine-103-dependent, PDZ-independent mechanism by which Amotl2 drives c-Src-mediated MAPK/ERK activation in endothelial morphogenesis.\",\n      \"evidence\": \"HUVEC siRNA, Y103 site-directed mutagenesis, ERK assays, and zebrafish vessel-growth readouts\",\n      \"pmids\": [\"21937427\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Y103 phosphorylation is regulated upstream unknown\", \"Relationship of ERK arm to junctional/Hippo functions not integrated\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed Amotl2 restrains Wnt signaling by physically trapping \\u03b2-catenin in recycling endosomes rather than acting at the destruction complex.\",\n      \"evidence\": \"Zebrafish epistasis with axin1 mutant, co-IP, and Rab11 endosome co-localization\",\n      \"pmids\": [\"22362771\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Determinants of \\u03b2-catenin capture vs release not defined\", \"Whether endosomal sequestration is regulated by signaling unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified AMOTL2 as a direct, Hippo-independent regulator of TAZ nuclear translocation via a PPXY-WW interaction, foreshadowing its central role over YAP/TAZ.\",\n      \"evidence\": \"Co-IP with domain mutagenesis (PPXY), luciferase reporter, and overexpression in H441 lung cells\",\n      \"pmids\": [\"23911299\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single cell type\", \"Did not reconcile Hippo-independent claim with later LATS-dependent mechanism\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Placed Amotl2 at specialized actin-based adhesion structures (synaptic podosomes, invadopodia) through interaction with the scaffold LL5\\u03b2.\",\n      \"evidence\": \"Affinity purification/MS, co-IP, and siRNA knockdown with morphometric analysis in myotubes and fibroblasts\",\n      \"pmids\": [\"23525008\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking LL5\\u03b2 binding to podosome size unknown\", \"Not connected to cadherin/Hippo functions\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated that AmotL2 mechanically couples VE-cadherin junctions to contractile actin to generate the tensile forces required for vascular lumen expansion.\",\n      \"evidence\": \"Zebrafish/mouse inactivation, endothelial co-IP with VE-cadherin complex, and traction force microscopy\",\n      \"pmids\": [\"24806444\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct actin-binding interface not resolved\", \"How force is sensed/transduced not yet defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed isoform-specific control whereby a hypoxia-induced p60 AmotL2 sequesters Crb3/Par3 polarity complexes to disrupt apical-basal polarity and drive invasion.\",\n      \"evidence\": \"c-Fos promoter analysis, co-IP with polarity complexes, vesicle trafficking assays, and xenograft\",\n      \"pmids\": [\"25080976\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Isoform-specific structural basis for polarity-complex retention unknown\", \"p60 vs p100 abundance control incompletely defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified mTORC2-dependent S760 phosphorylation as an off-switch for AMOTL2's YAP-repressive function, linking growth signaling to Hippo output.\",\n      \"evidence\": \"Mouse glioma gene-trap, S760A/S760E mutagenesis, co-IP with YAP, and xenograft tumor assays\",\n      \"pmids\": [\"25998128\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How S760 phosphorylation alters AMOTL2-YAP binding structurally unknown\", \"Other mTORC2 substrates in the axis not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Established Par3 as the localization determinant placing AmotL2 at junctions to build the cadherin-actin axis controlling tissue geometry.\",\n      \"evidence\": \"Zebrafish inactivation, cell-culture siRNA, immunofluorescence, and co-IP\",\n      \"pmids\": [\"28790366\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect Par3-AmotL2 interaction not distinguished\", \"Single lab\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed the p100 isoform forms an E-cadherin/radial-actin complex that generates the contractile tension required for blastocyst hatching and epithelial packing.\",\n      \"evidence\": \"Co-IP, zebrafish/mouse inactivation, and blebbistatin phenocopy of myosin-II loss\",\n      \"pmids\": [\"28842668\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of E-cadherin/AmotL2/actin complex unresolved\", \"Quantitative force contribution not measured\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Uncovered a junction-independent role for AMOTL2 in sequestering the PP2A subunit PPP2R2A in the cytoplasm to sustain phospho-JUN and AP-1 proliferation signaling.\",\n      \"evidence\": \"CRISPR loss-of-function screen, MS, co-IP/GST pull-down, fractionation, and phospho-JUN immunoblot in NSCLC\",\n      \"pmids\": [\"32950569\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PPP2R2A binding competes with YAP/cadherin functions unknown\", \"Regulation of this interaction not defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined WWP1-mediated mono-ubiquitination at K347/K408 as the molecular trigger that enables AMOTL2 to recruit LATS2/SAV1 and drive YAP phosphorylation at high cell density.\",\n      \"evidence\": \"In vitro ubiquitination, K347/K408 mutagenesis, co-IP with LATS2/SAV1, and contact-inhibition assays\",\n      \"pmids\": [\"34404733\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Deubiquitinase counteracting WWP1 not identified\", \"How Crumbs coupling is regulated mechanistically unresolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended the \\u03b2-catenin sequestration mechanism to glioma, showing AMOTL2 loss enhances Wnt-driven proliferation and invasion.\",\n      \"evidence\": \"Co-IP, immunofluorescence, and siRNA knockdown in glioma cell lines\",\n      \"pmids\": [\"34036399\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, no in vivo validation\", \"Did not test endosomal trapping specifically\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Placed AMOTL2 downstream of MAGI1 loss as an upstream driver of a ROCK/p38 stress pathway in breast cancer.\",\n      \"evidence\": \"Double-knockdown epistasis, ROCK/p38 inhibitor rescue, and cell-stiffness/tumorigenicity assays\",\n      \"pmids\": [\"33707576\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link from AMOTL2 to ROCK/p38 not shown\", \"In vitro only\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Confirmed AMOTL2 restrains TGF-\\u03b21-induced YAP1 activation in airway smooth muscle, linking it to proliferation and ECM deposition.\",\n      \"evidence\": \"Overexpression/knockdown in ASM cells with YAP1 nuclear-localization and constitutively-active YAP1 rescue\",\n      \"pmids\": [\"34323359\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct AMOTL2-YAP1 binding assay reported\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed AMOTL2 is transcriptionally repressed by MEF2D, derepressing YAP signaling to promote hepatocellular carcinoma.\",\n      \"evidence\": \"Dual luciferase reporter and overexpression in hepatoma cells\",\n      \"pmids\": [\"35698637\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Limited mechanistic depth; promoter binding inferred\", \"No in vivo confirmation\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified CLK2-dependent alternative splicing as a regulator that generates a membrane-uncoupled AMOTL2 isoform, switching off YAP repression.\",\n      \"evidence\": \"Chemical screen, RT-PCR splicing analysis, and YAP phosphorylation/localization assays with CLK2 inhibitor\",\n      \"pmids\": [\"38126343\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Spliced isoform binding partners not reconstituted\", \"Physiological context of CLK2 control unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed the mechanism of p60-driven invasion: p60 binds p100 AMOTL2 to uncouple E-cadherin-radial actin from the nuclear lamina, softening the nucleus for ameboid migration.\",\n      \"evidence\": \"Inter-isoform co-IP, atomic force microscopy of nuclear stiffness, and micropore invasion assays\",\n      \"pmids\": [\"37443716\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct nuclear-lamina binding partner not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrated in vivo that AmotL2 transmits flow-derived mechanical signals from VE-cadherin junctions through actin to the nuclear lamina, with endothelial loss causing aortic aneurysms.\",\n      \"evidence\": \"Endothelial-specific mouse knockout, tripartite co-IP, nuclear-lamina connectivity assays, and transcriptomics\",\n      \"pmids\": [\"39195920\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the nuclear-lamina anchoring partner unresolved\", \"Mechanotransduction signaling steps to inflammation not fully mapped\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified ARNTL2 as a transcriptional repressor of AMOTL2 that, by reducing LATS1/2 recruitment and stabilization, activates YAP to promote nasopharyngeal carcinoma invasion.\",\n      \"evidence\": \"ChIP, knockdown epistasis with rescue, LATS1/2 co-IP, and xenograft\",\n      \"pmids\": [\"38956029\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct LATS1/2 stabilization mechanism vs recruitment not separated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Positioned AMOTL2 as a mediator in a WBP2/c-JUN chemoresistance axis in breast cancer.\",\n      \"evidence\": \"RNA-seq, knockdown epistasis, phospho-c-JUN immunoblot, and xenograft with MG132\",\n      \"pmids\": [\"39709035\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct AMOTL2/c-JUN binding assay\", \"Mechanism of drug-resistance protein induction unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified AMOTL2 as a direct binding target of the natural product celastrol, linking pharmacological engagement to Hippo activation and cardiomyocyte apoptosis.\",\n      \"evidence\": \"Activity-based protein profiling, shRNA knockdown rescue, YAP1 phosphorylation and mitochondrial biogenesis assays, and mouse cardiotoxicity model\",\n      \"pmids\": [\"41412440\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Celastrol binding site on AMOTL2 not mapped\", \"Whether binding mimics a physiological regulatory event unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Uncovered an antiviral role in which AMOTL2 promotes the type I interferon response by modulating STAT1 to restrict Zika virus replication.\",\n      \"evidence\": \"Genome-wide CRISPR knockout screen for ZIKV host factors with STAT1 activation and ISG expression follow-up\",\n      \"pmids\": [\"40892926\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular interaction with STAT1 not demonstrated\", \"Relation to junctional/Hippo roles unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed AmotL2 and Yap1 form an opposing mechanosensitive module balancing actomyosin tension to enable flow-guided vascular pruning.\",\n      \"evidence\": \"Zebrafish live imaging, amotL2/yap1 genetic analysis, and actomyosin tension measurements (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.11.19.689183\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint not yet peer-reviewed\", \"Molecular basis of the AmotL2-Yap1 balance during pruning not resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis by which AMOTL2 physically bridges cadherin, actin, and the nuclear lamina, and how its multiple regulatory inputs (ubiquitination, phosphorylation, splicing, isoform balance, transcriptional repression) are integrated within a single cell, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of AMOTL2 complexes\", \"Nuclear-lamina anchoring partner unidentified\", \"Cross-talk between Hippo, Wnt, MAPK, and interferon roles not integrated\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 5, 9, 11, 18]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [5, 9, 18]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [7, 11, 19, 14]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [2, 6, 10]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 5, 8, 18]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3, 10]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [2, 6]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [5, 9, 18]},\n      {\"term_id\": \"GO:0005635\", \"supporting_discovery_ids\": [17, 18]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 7, 11, 19]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [5, 8, 9, 18]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [22]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [6, 10, 17]}\n    ],\n    \"complexes\": [\n      \"VE-cadherin adherens junction complex\",\n      \"E-cadherin/p100-AMOTL2/radial actin complex\",\n      \"LATS2/SAV1 Hippo complex\"\n    ],\n    \"partners\": [\n      \"CDH5\",\n      \"CDH1\",\n      \"TAZ/WWTR1\",\n      \"YAP1\",\n      \"LATS2\",\n      \"CTNNB1\",\n      \"PPP2R2A\",\n      \"PARD3\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}