{"gene":"AMOTL1","run_date":"2026-04-28T17:12:37","timeline":{"discoveries":[{"year":2001,"finding":"AMOTL1 (JEAP) is a novel peripheral membrane protein that localizes to tight junctions in exocrine cells, identified by fluorescence localization-based expression cloning. It contains a coiled-coil domain and a PDZ-binding motif at the C-terminus, and co-localizes with ZO-1 and occludin at tight junctions in polarized epithelial cells.","method":"Fluorescence localization-based expression cloning, immunofluorescence microscopy, co-localization with TJ markers","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment with defined subcellular phenotype, single lab","pmids":["11733531"],"is_preprint":false},{"year":2007,"finding":"AMOTL1 (JEAP) physically interacts with the multi-PDZ domain proteins MUPP1 and Patj via its C-terminal PDZ-binding motif (PDZ3 of MUPP1 responsible for JEAP interaction). AMOTL1 localizes to tight junctions and apical membranes as a peripheral membrane protein; however, the PDZ-binding motif is not strictly required for TJ localization.","method":"Yeast two-hybrid screening, immunofluorescence microscopy, biochemical fractionation, domain mapping","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 1-2 — yeast two-hybrid plus domain mapping plus fractionation and imaging, multiple orthogonal methods","pmids":["17397395"],"is_preprint":false},{"year":2014,"finding":"miR-124 represses vasculogenic mimicry, migration, and invasion in cervical cancer cells by targeting the 3'UTR of AMOTL1, thereby negatively regulating AMOTL1 expression and suppressing EMT.","method":"3'UTR luciferase reporter assay, miRNA overexpression, in vitro migration/invasion assays","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 3 — functional KD/OE with phenotype and 3'UTR reporter validation, single lab","pmids":["25218344"],"is_preprint":false},{"year":2016,"finding":"The E3 ubiquitin ligase HECW2 physically interacts with AMOTL1 and enhances its protein stability via K63-linked ubiquitination in endothelial cells. HECW2 depletion decreases AMOTL1 stability, loosens cell-to-cell junctions, and causes YAP to translocate from cytoplasm to nucleus, promoting angiogenic sprouting.","method":"Co-immunoprecipitation, ubiquitination assays (K63-linkage specific), siRNA knockdown, immunofluorescence, angiogenic sprouting assay","journal":"Cellular signalling","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP plus ubiquitination linkage determination plus KD phenotype with mechanistic follow-up","pmids":["27498087"],"is_preprint":false},{"year":2016,"finding":"The tumor suppressor Merlin triggers proteasomal degradation of AMOTL1 through direct interaction and recruitment of NEDD family ubiquitin ligases. In parallel, YAP stimulates AMOTL1 expression. AMOTL1 promotes tumor cell migration and proliferation by activating c-Src.","method":"Co-immunoprecipitation, proteasome inhibition assays, siRNA knockdown, cell migration assays, c-Src activity measurement","journal":"Neoplasia (New York, N.Y.)","confidence":"Medium","confidence_rationale":"Tier 2 — direct interaction demonstrated with mechanistic follow-up, single lab","pmids":["26806348"],"is_preprint":false},{"year":2017,"finding":"In the mouse heart, AMOTL1 acts downstream of the atypical cadherin Fat4 in a non-canonical Hippo pathway. Fat4 sequesters AMOTL1 out of the nucleus; loss of Fat4 leads to nuclear translocation of AMOTL1 together with YAP1, promoting cardiomyocyte proliferation and heart overgrowth. Fat4 is not required for canonical Hippo kinase activation.","method":"Mouse genetic knockout (Fat4 mutant), immunofluorescence for nuclear/cytoplasmic localization, cardiomyocyte proliferation assays, epistasis analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis in mouse model with defined cellular phenotype and localization data, multiple orthogonal methods","pmids":["28239148"],"is_preprint":false},{"year":2019,"finding":"circAMOTL1 acts as a competing endogenous RNA (ceRNA) by sponging miR-485-5p, thereby relieving miR-485-5p-mediated repression of AMOTL1 mRNA and increasing AMOTL1 protein levels to promote cervical cancer cell growth.","method":"qRT-PCR, gain/loss-of-function assays, luciferase reporter assay for ceRNA mechanism, in vivo xenograft","journal":"Molecular therapy. Nucleic acids","confidence":"Medium","confidence_rationale":"Tier 3 — ceRNA mechanism with rescue experiments, single lab","pmids":["31812104"],"is_preprint":false},{"year":2020,"finding":"AMOTL1 physically interacts with YAP1 in the cytoplasm via co-immunoprecipitation and immunofluorescence; this interaction protects both proteins from ubiquitin-mediated proteasomal degradation. AMOTL1 promotes YAP1 nuclear translocation to activate downstream targets CTGF and c-Myc in gastric cancer cells.","method":"Co-immunoprecipitation, immunofluorescence, ubiquitination assay, siRNA knockdown, xenograft assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — direct protein interaction with mechanistic follow-up (ubiquitination protection, nuclear translocation, downstream transcription), multiple orthogonal methods","pmids":["32313226"],"is_preprint":false},{"year":2023,"finding":"SRSF3 binds directly to exon 12 of AMOTL1 via its RRM domain to promote inclusion of exon 12, generating a long isoform (AMOTL1-L). AMOTL1-L preferentially localizes intracellularly (versus membrane localization of AMOTL1-S) and more robustly interacts with YAP1 to promote its nuclear translocation and NPC cell proliferation/migration.","method":"Transcriptome analysis, RNA-protein binding assay (RRM domain mapping), immunofluorescence for localization, co-immunoprecipitation, functional rescue assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 — direct RNA-protein interaction mapping plus functional isoform comparison, single lab","pmids":["37558679"],"is_preprint":false},{"year":2024,"finding":"N-acetyltransferase 10 (Nat10) mediates N4-acetylcytidine (ac4C) modification of Amotl1 mRNA, increasing its stability and translation in cardiac fibroblasts. This leads to increased AMOTL1 protein, enhanced interaction with YAP1, and facilitation of YAP1 nuclear translocation, driving cardiac fibroblast proliferation and myofibroblast differentiation after myocardial infarction.","method":"ac4C-RIP-seq, siRNA knockdown, fibroblast-specific Nat10 KO/OE mouse models, co-immunoprecipitation, verteporfin YAP inhibition","journal":"Acta pharmacologica Sinica","confidence":"High","confidence_rationale":"Tier 2 — ac4C-RIP-seq plus genetic KO/OE mouse model plus mechanistic follow-up in fibroblasts, multiple orthogonal methods","pmids":["38839936"],"is_preprint":false},{"year":2026,"finding":"AMOTL1 contains a Tankyrase-binding domain (TBD) encompassing residues R157 and P160. Disease-associated R157C and P160L mutations abolish interaction with Tankyrase 1/2 (TNKS1/2) and RNF146, preventing PARylation, ubiquitination, and proteasomal degradation of AMOTL1. These stabilized mutants accumulate in the cytoplasm, disrupt cell junctions and focal adhesions, and impair cell migration velocity and persistence. In zebrafish, R157C expression causes craniofacial malformations and cardiac/skeletal muscle defects.","method":"Co-immunoprecipitation, ubiquitination assay, PARylation assay, live-cell imaging of migration, zebrafish embryo expression system","journal":"Bioscience reports","confidence":"High","confidence_rationale":"Tier 2 — direct interaction mapping with multiple biochemical assays plus in vivo zebrafish validation, multiple orthogonal methods","pmids":["42012498"],"is_preprint":false},{"year":2026,"finding":"AMOTL1 contains three PPxY motifs that engage WW-domain proteins NEDD4-1 and KIBRA through distinct cooperative binding mechanisms. NEDD4-1 simultaneously engages all three PPxY motifs with three of its four WW domains, producing ~10-fold enhanced affinity (promoting AMOTL1 degradation). KIBRA binds primarily via the C-terminal PPxY motif with high affinity, with transient secondary contacts that do not enhance overall binding (protecting AMOTL1 from degradation).","method":"Isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR) spectroscopy, quantitative molecular biophysical analyses","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 — in vitro biophysical reconstitution with ITC and NMR, mechanistic detail on cooperativity","pmids":["41580069"],"is_preprint":false},{"year":2026,"finding":"PFKP (platelet-type phosphofructokinase) directly binds AMOTL1 and inhibits its ubiquitin-mediated proteasomal degradation. PFKP-driven aerobic glycolysis and EMT in head and neck cancer cells are AMOTL1-dependent. PFKP promotes YAP nuclear translocation via AMOTL1, suppressing Hippo pathway activity.","method":"Co-immunoprecipitation, ubiquitination analysis, immunofluorescence, siRNA knockdown functional rescue, in vivo nude mouse tumor model","journal":"Journal of translational internal medicine","confidence":"Medium","confidence_rationale":"Tier 2 — direct binding with ubiquitination and functional rescue assays, single lab","pmids":["41727965"],"is_preprint":false},{"year":2025,"finding":"Tankyrase inhibition (by OM-153) stabilizes AMOTL1 protein, consistent with AMOTL1 being a direct substrate of Tankyrase-mediated regulation, and this suppresses YAP signaling and reduces pro-fibrotic ECM expression in preclinical IPF models.","method":"Tankyrase inhibitor treatment in primary lung fibroblasts, lung-on-a-chip, precision-cut lung slices, bleomycin mouse model; immunoblotting and qRT-PCR","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 3 — pharmacological inhibition in multiple preclinical models, preprint, no mutagenesis confirmation","pmids":["bio_10.1101_2025.11.13.688191"],"is_preprint":true}],"current_model":"AMOTL1 is a scaffold/peripheral membrane protein that localizes to tight junctions and the cytoplasm, where it interacts with YAP1 to protect it from ubiquitin-mediated degradation and promotes its nuclear translocation; AMOTL1 protein levels are regulated by multiple E3 ligase complexes (NEDD4-1 via PPxY–WW domain interactions, RNF146/TNKS1/2 via PARylation of its TBD, and Merlin-recruited NEDD family ligases) and stabilized by HECW2-mediated K63-linked ubiquitination, placing AMOTL1 as a non-canonical Hippo pathway intermediate that bridges upstream signals (Fat4, Tankyrase, Merlin, PFKP) to YAP1-driven transcription and thereby controls cell migration, junction integrity, proliferation, and organ growth."},"narrative":{"teleology":[{"year":2001,"claim":"Identification of AMOTL1 as a peripheral membrane protein at tight junctions established it as a component of the junctional machinery rather than a transmembrane receptor, raising the question of how it is recruited and what signaling it mediates.","evidence":"Fluorescence localization-based expression cloning and co-localization with ZO-1/occludin in polarized epithelial cells","pmids":["11733531"],"confidence":"Medium","gaps":["No functional perturbation performed","Mechanism of membrane recruitment not determined","No signaling pathway linkage established"]},{"year":2007,"claim":"Discovery that AMOTL1 binds the multi-PDZ proteins MUPP1 and Patj via its C-terminal PDZ-binding motif—yet this motif is dispensable for tight-junction localization—revealed that AMOTL1 uses distinct determinants for localization versus protein–protein interactions at junctions.","evidence":"Yeast two-hybrid screening, domain mapping, immunofluorescence, and biochemical fractionation","pmids":["17397395"],"confidence":"High","gaps":["No loss-of-function phenotype assessed","Other localization determinants not mapped","Functional consequence of MUPP1/Patj binding unknown"]},{"year":2016,"claim":"Two independent studies revealed that AMOTL1 stability is regulated by opposing ubiquitin signals—HECW2 stabilizes it via K63-linked ubiquitination while Merlin promotes its proteasomal degradation via NEDD-family ligases—establishing AMOTL1 protein turnover as a signaling integration point controlling YAP localization and cell migration.","evidence":"Reciprocal Co-IP, K63-linkage-specific ubiquitination assays, siRNA knockdown with junction/angiogenesis phenotypes (HECW2 study); Co-IP, proteasome inhibition, c-Src activity measurement (Merlin study)","pmids":["27498087","26806348"],"confidence":"High","gaps":["Whether HECW2 and Merlin pathways converge on the same ubiquitination sites is unknown","In vivo genetic validation of HECW2–AMOTL1 axis not performed","Structural basis of Merlin–AMOTL1 interaction not determined"]},{"year":2017,"claim":"Genetic epistasis in the mouse heart showed that Fat4 sequesters AMOTL1 out of the nucleus; loss of Fat4 causes AMOTL1–YAP1 nuclear co-translocation and cardiomyocyte overgrowth, placing AMOTL1 as a non-canonical Hippo pathway intermediate downstream of Fat4 and independent of canonical Hippo kinases.","evidence":"Fat4 knockout mouse, immunofluorescence for nuclear/cytoplasmic localization, cardiomyocyte proliferation assays, epistasis analysis","pmids":["28239148"],"confidence":"High","gaps":["Direct physical interaction between Fat4 and AMOTL1 not biochemically demonstrated","Whether this pathway operates outside the heart is unknown","Mechanism by which Fat4 retains AMOTL1 cytoplasmically not resolved"]},{"year":2020,"claim":"Demonstration that AMOTL1 physically binds YAP1, reciprocally protects both proteins from proteasomal degradation, and promotes YAP1 nuclear translocation to activate CTGF and c-Myc established the core biochemical mechanism linking AMOTL1 to transcriptional output.","evidence":"Co-immunoprecipitation, ubiquitination assay, immunofluorescence, siRNA knockdown, xenograft assay in gastric cancer cells","pmids":["32313226"],"confidence":"High","gaps":["Identity of the E3 ligase targeting the AMOTL1–YAP1 complex is not specified","Whether AMOTL1 enters the nucleus with YAP1 or acts at nuclear pores is unresolved","Structural basis of AMOTL1–YAP1 interaction not determined"]},{"year":2023,"claim":"Identification of SRSF3-regulated alternative splicing producing a long AMOTL1 isoform (AMOTL1-L) that preferentially localizes intracellularly and more robustly promotes YAP1 nuclear translocation revealed that isoform-specific regulation tunes AMOTL1's signaling output.","evidence":"RNA-protein binding assay with RRM domain mapping, immunofluorescence for isoform localization, Co-IP, functional rescue in NPC cells","pmids":["37558679"],"confidence":"Medium","gaps":["Structural difference between AMOTL1-L and AMOTL1-S explaining differential localization not resolved","Relative abundance of isoforms across tissues not surveyed","Single-lab finding awaiting independent replication"]},{"year":2024,"claim":"Nat10-mediated ac4C modification of Amotl1 mRNA was shown to increase its stability and translation, connecting an epitranscriptomic input to AMOTL1 protein levels and downstream YAP1 signaling in cardiac fibroblasts after myocardial infarction.","evidence":"ac4C-RIP-seq, fibroblast-specific Nat10 KO/OE mouse models, Co-IP, verteporfin YAP inhibition","pmids":["38839936"],"confidence":"High","gaps":["Specific ac4C sites on Amotl1 mRNA not mapped at nucleotide resolution","Whether ac4C regulation of AMOTL1 extends beyond cardiac fibroblasts is unknown","Contribution relative to miRNA-mediated regulation not compared"]},{"year":2026,"claim":"Biophysical reconstitution of AMOTL1's three PPxY motifs with NEDD4-1 and KIBRA WW domains revealed that cooperative multivalent engagement by NEDD4-1 (10-fold affinity enhancement) versus single-site binding by KIBRA explains how competing E3 ligase and protector complexes differentially regulate AMOTL1 stability.","evidence":"Isothermal titration calorimetry (ITC) and NMR spectroscopy with purified proteins","pmids":["41580069"],"confidence":"High","gaps":["Cellular validation of cooperativity-dependent degradation rates not performed","Whether post-translational modifications modulate WW–PPxY affinities in vivo is unknown","KIBRA's protective effect not confirmed by in vivo genetic experiment"]},{"year":2026,"claim":"Mapping of the Tankyrase-binding domain and identification of disease-associated R157C/P160L mutations that abolish TNKS1/2 and RNF146 binding, PARylation, and degradation established a direct PARylation-dependent degradation axis; stabilized mutants disrupt junctions and cause craniofacial and cardiac defects in zebrafish.","evidence":"Co-IP, PARylation and ubiquitination assays, live-cell migration imaging, zebrafish embryo expression","pmids":["42012498"],"confidence":"High","gaps":["Human genetic disease association not confirmed by family segregation or GWAS","Whether R157C/P160L mutations affect NEDD4-1/KIBRA axis is untested","Rescue of zebrafish phenotype by Tankyrase overexpression not attempted"]},{"year":2026,"claim":"Discovery that PFKP directly binds and stabilizes AMOTL1 to promote YAP nuclear translocation linked glycolytic metabolism to Hippo pathway suppression via AMOTL1, explaining how metabolic reprogramming drives EMT in head and neck cancer.","evidence":"Co-IP, ubiquitination analysis, siRNA functional rescue, nude mouse xenograft","pmids":["41727965"],"confidence":"Medium","gaps":["Mechanism by which PFKP inhibits AMOTL1 ubiquitination is not defined","Single-lab finding awaiting independent replication","Whether PFKP enzymatic activity or scaffolding function is required is unresolved"]},{"year":null,"claim":"The structural basis of the AMOTL1–YAP1 interaction, the identity of the E3 ligase(s) targeting this complex, and whether AMOTL1 enters the nucleus as a co-factor or acts solely at the cytoplasmic-nuclear boundary remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No atomic-resolution structure of AMOTL1 or AMOTL1–YAP1 complex","Nuclear versus cytoplasmic function of AMOTL1 not genetically separated","Integration of multiple degradation axes (Tankyrase, NEDD4-1, Merlin, HECW2, PFKP, KIBRA) into a unified quantitative model not achieved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[5,7,12]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[7,5]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[7,8,10]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[5,7,8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[5,7,9,12]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[0,1,3,10]}],"complexes":[],"partners":["YAP1","NEDD4","KIBRA","TNKS1","RNF146","HECW2","NF2","PFKP"],"other_free_text":[]},"mechanistic_narrative":"AMOTL1 is a scaffold protein that operates as a non-canonical intermediate in the Hippo signaling pathway, integrating upstream signals from tight junctions, cadherins, and metabolic regulators to control YAP1-dependent transcription, cell proliferation, migration, and organ growth. AMOTL1 physically binds YAP1 in the cytoplasm, protecting it from ubiquitin-mediated degradation and facilitating its nuclear translocation to activate targets including CTGF and c-Myc; in the developing heart, the atypical cadherin Fat4 sequesters AMOTL1 from the nucleus, and loss of Fat4 causes AMOTL1–YAP1 co-translocation and cardiomyocyte overgrowth [PMID:32313226, PMID:28239148]. AMOTL1 protein turnover is tightly regulated by multiple E3 ligase systems: NEDD4-1 cooperatively engages its three PPxY motifs via WW domains to promote degradation, Tankyrase 1/2 PARylates its Tankyrase-binding domain enabling RNF146-mediated ubiquitination, and Merlin recruits NEDD-family ligases for proteasomal destruction, while HECW2 stabilizes AMOTL1 through K63-linked ubiquitination and KIBRA competes with NEDD4-1 at the PPxY motifs to protect it [PMID:41580069, PMID:42012498, PMID:27498087, PMID:26806348]. Disease-associated mutations (R157C, P160L) in the Tankyrase-binding domain abolish PARylation-dependent degradation, causing AMOTL1 accumulation that disrupts cell junctions and focal adhesions and produces craniofacial and cardiac defects in zebrafish [PMID:42012498]."},"prefetch_data":{"uniprot":{"accession":"Q8IY63","full_name":"Angiomotin-like protein 1","aliases":[],"length_aa":956,"mass_kda":106.6,"function":"Inhibits the Wnt/beta-catenin signaling pathway, probably by recruiting CTNNB1 to recycling endosomes and hence preventing its translocation to the nucleus","subcellular_location":"Cell junction, tight junction","url":"https://www.uniprot.org/uniprotkb/Q8IY63/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMOTL1","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":"DYNLL1","stoichiometry":0.2},{"gene":"DYNLL2","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/AMOTL1","total_profiled":1310},"omim":[{"mim_id":"621192","title":"CRANIOFACIOCARDIOHEPATIC SYNDROME; CFCHS","url":"https://www.omim.org/entry/621192"},{"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":"610396","title":"TRAFFICKING PROTEIN PARTICLE COMPLEX, SUBUNIT 6A; TRAPPC6A","url":"https://www.omim.org/entry/610396"},{"mim_id":"300410","title":"ANGIOMOTIN; AMOT","url":"https://www.omim.org/entry/300410"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Cell Junctions","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":104.6}],"url":"https://www.proteinatlas.org/search/AMOTL1"},"hgnc":{"alias_symbol":["JEAP"],"prev_symbol":[]},"alphafold":{"accession":"Q8IY63","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IY63","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IY63-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IY63-F1-predicted_aligned_error_v6.png","plddt_mean":59.91},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AMOTL1","jax_strain_url":"https://www.jax.org/strain/search?query=AMOTL1"},"sequence":{"accession":"Q8IY63","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IY63.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IY63/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IY63"}},"corpus_meta":[{"pmid":"25218344","id":"PMC_25218344","title":"MiR-124 represses vasculogenic mimicry and cell motility by targeting amotL1 in cervical cancer cells.","date":"2014","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/25218344","citation_count":83,"is_preprint":false},{"pmid":"17397395","id":"PMC_17397395","title":"Molecular characterization of angiomotin/JEAP family proteins: interaction with MUPP1/Patj and their endogenous properties.","date":"2007","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/17397395","citation_count":81,"is_preprint":false},{"pmid":"28239148","id":"PMC_28239148","title":"Amotl1 mediates sequestration of the Hippo effector Yap1 downstream of Fat4 to restrict heart growth.","date":"2017","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/28239148","citation_count":81,"is_preprint":false},{"pmid":"31812104","id":"PMC_31812104","title":"circAMOTL1 Motivates AMOTL1 Expression to Facilitate Cervical Cancer Growth.","date":"2019","source":"Molecular therapy. Nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/31812104","citation_count":73,"is_preprint":false},{"pmid":"11733531","id":"PMC_11733531","title":"JEAP, a novel component of tight junctions in exocrine cells.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11733531","citation_count":58,"is_preprint":false},{"pmid":"27498087","id":"PMC_27498087","title":"The endothelial E3 ligase HECW2 promotes endothelial cell junctions by increasing AMOTL1 protein stability via K63-linked ubiquitination.","date":"2016","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/27498087","citation_count":43,"is_preprint":false},{"pmid":"32313226","id":"PMC_32313226","title":"AMOTL1 enhances YAP1 stability and promotes YAP1-driven gastric oncogenesis.","date":"2020","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/32313226","citation_count":37,"is_preprint":false},{"pmid":"26806348","id":"PMC_26806348","title":"AMOTL1 Promotes Breast Cancer Progression and Is Antagonized by Merlin.","date":"2016","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/26806348","citation_count":32,"is_preprint":false},{"pmid":"32884340","id":"PMC_32884340","title":"The circ-AMOTL1/ENO1 Axis Implicated in the Tumorigenesis of OLP-Associated Oral Squamous Cell Carcinoma.","date":"2020","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/32884340","citation_count":17,"is_preprint":false},{"pmid":"37586467","id":"PMC_37586467","title":"Circ-AMOTL1 enhances cardiac fibrosis through binding with EIF4A3 and stabilizing MARCKS expression in diabetic cardiomyopathy.","date":"2023","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/37586467","citation_count":13,"is_preprint":false},{"pmid":"36768425","id":"PMC_36768425","title":"CircAMOTL1 RNA and AMOTL1 Protein: Complex Functions of AMOTL1 Gene Products.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36768425","citation_count":8,"is_preprint":false},{"pmid":"37558679","id":"PMC_37558679","title":"SRSF3/AMOTL1 splicing axis promotes the tumorigenesis of nasopharyngeal carcinoma through regulating the nucleus translocation of YAP1.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37558679","citation_count":7,"is_preprint":false},{"pmid":"38839936","id":"PMC_38839936","title":"Acetylcytidine modification of Amotl1 by N-acetyltransferase 10 contributes to cardiac fibrotic expansion in mice after myocardial infarction.","date":"2024","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/38839936","citation_count":6,"is_preprint":false},{"pmid":"38318480","id":"PMC_38318480","title":"circ-Amotl1 in extracellular vesicles derived from ADSCs improves wound healing by upregulating SPARC translation.","date":"2024","source":"Regenerative therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38318480","citation_count":5,"is_preprint":false},{"pmid":"36751037","id":"PMC_36751037","title":"A mutational hotspot in AMOTL1 defines a new syndrome of orofacial clefting, cardiac anomalies, and tall stature.","date":"2023","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/36751037","citation_count":4,"is_preprint":false},{"pmid":"38018210","id":"PMC_38018210","title":"Hsa_circ_0001278 Facilitates Colorectal Cancer Progression via Sponging miR-338-5p and Regulating AMOTL1 Expression.","date":"2025","source":"Combinatorial chemistry & high throughput screening","url":"https://pubmed.ncbi.nlm.nih.gov/38018210","citation_count":3,"is_preprint":false},{"pmid":"39643184","id":"PMC_39643184","title":"Harnessing machine learning and multi-omics to explore tumor evolutionary characteristics and the role of AMOTL1 in prostate cancer.","date":"2024","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/39643184","citation_count":2,"is_preprint":false},{"pmid":"36116699","id":"PMC_36116699","title":"Clinical report: Chronic liver dysfunction in an individual with an AMOTL1 variant.","date":"2022","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36116699","citation_count":1,"is_preprint":false},{"pmid":"42012498","id":"PMC_42012498","title":"Patients-derived AMOTL1 mutations lead to defective cell migration and tissue development.","date":"2026","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/42012498","citation_count":0,"is_preprint":false},{"pmid":"41580069","id":"PMC_41580069","title":"Multivalent AMOTL1 Engages NEDD4-1 and KIBRA Through Distinct Cooperative Binding Mechanisms.","date":"2026","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/41580069","citation_count":0,"is_preprint":false},{"pmid":"41727965","id":"PMC_41727965","title":"PFKP binding AMOTL1 promotes tumor aerobic glycolysis and epithelial-mesenchymal transition by modulating Hippo pathway in head and neck cancer.","date":"2026","source":"Journal of translational internal medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41727965","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.11.13.688191","title":"Tankyrase inhibition demonstrates anti-fibrotic effects in preclinical pulmonary fibrosis models","date":"2025-11-13","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.13.688191","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12325,"output_tokens":3587,"usd":0.04539},"stage2":{"model":"claude-opus-4-6","input_tokens":6970,"output_tokens":3225,"usd":0.173212},"total_usd":0.218602,"stage1_batch_id":"msgbatch_01TqNM6p6FsoMBT2jzQTwKBr","stage2_batch_id":"msgbatch_01Ya473eco8sTHbvRymQaU8o","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"AMOTL1 (JEAP) is a novel peripheral membrane protein that localizes to tight junctions in exocrine cells, identified by fluorescence localization-based expression cloning. It contains a coiled-coil domain and a PDZ-binding motif at the C-terminus, and co-localizes with ZO-1 and occludin at tight junctions in polarized epithelial cells.\",\n      \"method\": \"Fluorescence localization-based expression cloning, immunofluorescence microscopy, co-localization with TJ markers\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment with defined subcellular phenotype, single lab\",\n      \"pmids\": [\"11733531\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"AMOTL1 (JEAP) physically interacts with the multi-PDZ domain proteins MUPP1 and Patj via its C-terminal PDZ-binding motif (PDZ3 of MUPP1 responsible for JEAP interaction). AMOTL1 localizes to tight junctions and apical membranes as a peripheral membrane protein; however, the PDZ-binding motif is not strictly required for TJ localization.\",\n      \"method\": \"Yeast two-hybrid screening, immunofluorescence microscopy, biochemical fractionation, domain mapping\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — yeast two-hybrid plus domain mapping plus fractionation and imaging, multiple orthogonal methods\",\n      \"pmids\": [\"17397395\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"miR-124 represses vasculogenic mimicry, migration, and invasion in cervical cancer cells by targeting the 3'UTR of AMOTL1, thereby negatively regulating AMOTL1 expression and suppressing EMT.\",\n      \"method\": \"3'UTR luciferase reporter assay, miRNA overexpression, in vitro migration/invasion assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — functional KD/OE with phenotype and 3'UTR reporter validation, single lab\",\n      \"pmids\": [\"25218344\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The E3 ubiquitin ligase HECW2 physically interacts with AMOTL1 and enhances its protein stability via K63-linked ubiquitination in endothelial cells. HECW2 depletion decreases AMOTL1 stability, loosens cell-to-cell junctions, and causes YAP to translocate from cytoplasm to nucleus, promoting angiogenic sprouting.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays (K63-linkage specific), siRNA knockdown, immunofluorescence, angiogenic sprouting assay\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP plus ubiquitination linkage determination plus KD phenotype with mechanistic follow-up\",\n      \"pmids\": [\"27498087\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The tumor suppressor Merlin triggers proteasomal degradation of AMOTL1 through direct interaction and recruitment of NEDD family ubiquitin ligases. In parallel, YAP stimulates AMOTL1 expression. AMOTL1 promotes tumor cell migration and proliferation by activating c-Src.\",\n      \"method\": \"Co-immunoprecipitation, proteasome inhibition assays, siRNA knockdown, cell migration assays, c-Src activity measurement\",\n      \"journal\": \"Neoplasia (New York, N.Y.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct interaction demonstrated with mechanistic follow-up, single lab\",\n      \"pmids\": [\"26806348\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"In the mouse heart, AMOTL1 acts downstream of the atypical cadherin Fat4 in a non-canonical Hippo pathway. Fat4 sequesters AMOTL1 out of the nucleus; loss of Fat4 leads to nuclear translocation of AMOTL1 together with YAP1, promoting cardiomyocyte proliferation and heart overgrowth. Fat4 is not required for canonical Hippo kinase activation.\",\n      \"method\": \"Mouse genetic knockout (Fat4 mutant), immunofluorescence for nuclear/cytoplasmic localization, cardiomyocyte proliferation assays, epistasis analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis in mouse model with defined cellular phenotype and localization data, multiple orthogonal methods\",\n      \"pmids\": [\"28239148\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"circAMOTL1 acts as a competing endogenous RNA (ceRNA) by sponging miR-485-5p, thereby relieving miR-485-5p-mediated repression of AMOTL1 mRNA and increasing AMOTL1 protein levels to promote cervical cancer cell growth.\",\n      \"method\": \"qRT-PCR, gain/loss-of-function assays, luciferase reporter assay for ceRNA mechanism, in vivo xenograft\",\n      \"journal\": \"Molecular therapy. Nucleic acids\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — ceRNA mechanism with rescue experiments, single lab\",\n      \"pmids\": [\"31812104\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AMOTL1 physically interacts with YAP1 in the cytoplasm via co-immunoprecipitation and immunofluorescence; this interaction protects both proteins from ubiquitin-mediated proteasomal degradation. AMOTL1 promotes YAP1 nuclear translocation to activate downstream targets CTGF and c-Myc in gastric cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, ubiquitination assay, siRNA knockdown, xenograft assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct protein interaction with mechanistic follow-up (ubiquitination protection, nuclear translocation, downstream transcription), multiple orthogonal methods\",\n      \"pmids\": [\"32313226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"SRSF3 binds directly to exon 12 of AMOTL1 via its RRM domain to promote inclusion of exon 12, generating a long isoform (AMOTL1-L). AMOTL1-L preferentially localizes intracellularly (versus membrane localization of AMOTL1-S) and more robustly interacts with YAP1 to promote its nuclear translocation and NPC cell proliferation/migration.\",\n      \"method\": \"Transcriptome analysis, RNA-protein binding assay (RRM domain mapping), immunofluorescence for localization, co-immunoprecipitation, functional rescue assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct RNA-protein interaction mapping plus functional isoform comparison, single lab\",\n      \"pmids\": [\"37558679\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"N-acetyltransferase 10 (Nat10) mediates N4-acetylcytidine (ac4C) modification of Amotl1 mRNA, increasing its stability and translation in cardiac fibroblasts. This leads to increased AMOTL1 protein, enhanced interaction with YAP1, and facilitation of YAP1 nuclear translocation, driving cardiac fibroblast proliferation and myofibroblast differentiation after myocardial infarction.\",\n      \"method\": \"ac4C-RIP-seq, siRNA knockdown, fibroblast-specific Nat10 KO/OE mouse models, co-immunoprecipitation, verteporfin YAP inhibition\",\n      \"journal\": \"Acta pharmacologica Sinica\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ac4C-RIP-seq plus genetic KO/OE mouse model plus mechanistic follow-up in fibroblasts, multiple orthogonal methods\",\n      \"pmids\": [\"38839936\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"AMOTL1 contains a Tankyrase-binding domain (TBD) encompassing residues R157 and P160. Disease-associated R157C and P160L mutations abolish interaction with Tankyrase 1/2 (TNKS1/2) and RNF146, preventing PARylation, ubiquitination, and proteasomal degradation of AMOTL1. These stabilized mutants accumulate in the cytoplasm, disrupt cell junctions and focal adhesions, and impair cell migration velocity and persistence. In zebrafish, R157C expression causes craniofacial malformations and cardiac/skeletal muscle defects.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, PARylation assay, live-cell imaging of migration, zebrafish embryo expression system\",\n      \"journal\": \"Bioscience reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct interaction mapping with multiple biochemical assays plus in vivo zebrafish validation, multiple orthogonal methods\",\n      \"pmids\": [\"42012498\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"AMOTL1 contains three PPxY motifs that engage WW-domain proteins NEDD4-1 and KIBRA through distinct cooperative binding mechanisms. NEDD4-1 simultaneously engages all three PPxY motifs with three of its four WW domains, producing ~10-fold enhanced affinity (promoting AMOTL1 degradation). KIBRA binds primarily via the C-terminal PPxY motif with high affinity, with transient secondary contacts that do not enhance overall binding (protecting AMOTL1 from degradation).\",\n      \"method\": \"Isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR) spectroscopy, quantitative molecular biophysical analyses\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro biophysical reconstitution with ITC and NMR, mechanistic detail on cooperativity\",\n      \"pmids\": [\"41580069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"PFKP (platelet-type phosphofructokinase) directly binds AMOTL1 and inhibits its ubiquitin-mediated proteasomal degradation. PFKP-driven aerobic glycolysis and EMT in head and neck cancer cells are AMOTL1-dependent. PFKP promotes YAP nuclear translocation via AMOTL1, suppressing Hippo pathway activity.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination analysis, immunofluorescence, siRNA knockdown functional rescue, in vivo nude mouse tumor model\",\n      \"journal\": \"Journal of translational internal medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct binding with ubiquitination and functional rescue assays, single lab\",\n      \"pmids\": [\"41727965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Tankyrase inhibition (by OM-153) stabilizes AMOTL1 protein, consistent with AMOTL1 being a direct substrate of Tankyrase-mediated regulation, and this suppresses YAP signaling and reduces pro-fibrotic ECM expression in preclinical IPF models.\",\n      \"method\": \"Tankyrase inhibitor treatment in primary lung fibroblasts, lung-on-a-chip, precision-cut lung slices, bleomycin mouse model; immunoblotting and qRT-PCR\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — pharmacological inhibition in multiple preclinical models, preprint, no mutagenesis confirmation\",\n      \"pmids\": [\"bio_10.1101_2025.11.13.688191\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"AMOTL1 is a scaffold/peripheral membrane protein that localizes to tight junctions and the cytoplasm, where it interacts with YAP1 to protect it from ubiquitin-mediated degradation and promotes its nuclear translocation; AMOTL1 protein levels are regulated by multiple E3 ligase complexes (NEDD4-1 via PPxY–WW domain interactions, RNF146/TNKS1/2 via PARylation of its TBD, and Merlin-recruited NEDD family ligases) and stabilized by HECW2-mediated K63-linked ubiquitination, placing AMOTL1 as a non-canonical Hippo pathway intermediate that bridges upstream signals (Fat4, Tankyrase, Merlin, PFKP) to YAP1-driven transcription and thereby controls cell migration, junction integrity, proliferation, and organ growth.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"AMOTL1 is a scaffold protein that operates as a non-canonical intermediate in the Hippo signaling pathway, integrating upstream signals from tight junctions, cadherins, and metabolic regulators to control YAP1-dependent transcription, cell proliferation, migration, and organ growth. AMOTL1 physically binds YAP1 in the cytoplasm, protecting it from ubiquitin-mediated degradation and facilitating its nuclear translocation to activate targets including CTGF and c-Myc; in the developing heart, the atypical cadherin Fat4 sequesters AMOTL1 from the nucleus, and loss of Fat4 causes AMOTL1–YAP1 co-translocation and cardiomyocyte overgrowth [PMID:32313226, PMID:28239148]. AMOTL1 protein turnover is tightly regulated by multiple E3 ligase systems: NEDD4-1 cooperatively engages its three PPxY motifs via WW domains to promote degradation, Tankyrase 1/2 PARylates its Tankyrase-binding domain enabling RNF146-mediated ubiquitination, and Merlin recruits NEDD-family ligases for proteasomal destruction, while HECW2 stabilizes AMOTL1 through K63-linked ubiquitination and KIBRA competes with NEDD4-1 at the PPxY motifs to protect it [PMID:41580069, PMID:42012498, PMID:27498087, PMID:26806348]. Disease-associated mutations (R157C, P160L) in the Tankyrase-binding domain abolish PARylation-dependent degradation, causing AMOTL1 accumulation that disrupts cell junctions and focal adhesions and produces craniofacial and cardiac defects in zebrafish [PMID:42012498].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Identification of AMOTL1 as a peripheral membrane protein at tight junctions established it as a component of the junctional machinery rather than a transmembrane receptor, raising the question of how it is recruited and what signaling it mediates.\",\n      \"evidence\": \"Fluorescence localization-based expression cloning and co-localization with ZO-1/occludin in polarized epithelial cells\",\n      \"pmids\": [\"11733531\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional perturbation performed\", \"Mechanism of membrane recruitment not determined\", \"No signaling pathway linkage established\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Discovery that AMOTL1 binds the multi-PDZ proteins MUPP1 and Patj via its C-terminal PDZ-binding motif—yet this motif is dispensable for tight-junction localization—revealed that AMOTL1 uses distinct determinants for localization versus protein–protein interactions at junctions.\",\n      \"evidence\": \"Yeast two-hybrid screening, domain mapping, immunofluorescence, and biochemical fractionation\",\n      \"pmids\": [\"17397395\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No loss-of-function phenotype assessed\", \"Other localization determinants not mapped\", \"Functional consequence of MUPP1/Patj binding unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Two independent studies revealed that AMOTL1 stability is regulated by opposing ubiquitin signals—HECW2 stabilizes it via K63-linked ubiquitination while Merlin promotes its proteasomal degradation via NEDD-family ligases—establishing AMOTL1 protein turnover as a signaling integration point controlling YAP localization and cell migration.\",\n      \"evidence\": \"Reciprocal Co-IP, K63-linkage-specific ubiquitination assays, siRNA knockdown with junction/angiogenesis phenotypes (HECW2 study); Co-IP, proteasome inhibition, c-Src activity measurement (Merlin study)\",\n      \"pmids\": [\"27498087\", \"26806348\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether HECW2 and Merlin pathways converge on the same ubiquitination sites is unknown\", \"In vivo genetic validation of HECW2–AMOTL1 axis not performed\", \"Structural basis of Merlin–AMOTL1 interaction not determined\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Genetic epistasis in the mouse heart showed that Fat4 sequesters AMOTL1 out of the nucleus; loss of Fat4 causes AMOTL1–YAP1 nuclear co-translocation and cardiomyocyte overgrowth, placing AMOTL1 as a non-canonical Hippo pathway intermediate downstream of Fat4 and independent of canonical Hippo kinases.\",\n      \"evidence\": \"Fat4 knockout mouse, immunofluorescence for nuclear/cytoplasmic localization, cardiomyocyte proliferation assays, epistasis analysis\",\n      \"pmids\": [\"28239148\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct physical interaction between Fat4 and AMOTL1 not biochemically demonstrated\", \"Whether this pathway operates outside the heart is unknown\", \"Mechanism by which Fat4 retains AMOTL1 cytoplasmically not resolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstration that AMOTL1 physically binds YAP1, reciprocally protects both proteins from proteasomal degradation, and promotes YAP1 nuclear translocation to activate CTGF and c-Myc established the core biochemical mechanism linking AMOTL1 to transcriptional output.\",\n      \"evidence\": \"Co-immunoprecipitation, ubiquitination assay, immunofluorescence, siRNA knockdown, xenograft assay in gastric cancer cells\",\n      \"pmids\": [\"32313226\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the E3 ligase targeting the AMOTL1–YAP1 complex is not specified\", \"Whether AMOTL1 enters the nucleus with YAP1 or acts at nuclear pores is unresolved\", \"Structural basis of AMOTL1–YAP1 interaction not determined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of SRSF3-regulated alternative splicing producing a long AMOTL1 isoform (AMOTL1-L) that preferentially localizes intracellularly and more robustly promotes YAP1 nuclear translocation revealed that isoform-specific regulation tunes AMOTL1's signaling output.\",\n      \"evidence\": \"RNA-protein binding assay with RRM domain mapping, immunofluorescence for isoform localization, Co-IP, functional rescue in NPC cells\",\n      \"pmids\": [\"37558679\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural difference between AMOTL1-L and AMOTL1-S explaining differential localization not resolved\", \"Relative abundance of isoforms across tissues not surveyed\", \"Single-lab finding awaiting independent replication\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Nat10-mediated ac4C modification of Amotl1 mRNA was shown to increase its stability and translation, connecting an epitranscriptomic input to AMOTL1 protein levels and downstream YAP1 signaling in cardiac fibroblasts after myocardial infarction.\",\n      \"evidence\": \"ac4C-RIP-seq, fibroblast-specific Nat10 KO/OE mouse models, Co-IP, verteporfin YAP inhibition\",\n      \"pmids\": [\"38839936\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific ac4C sites on Amotl1 mRNA not mapped at nucleotide resolution\", \"Whether ac4C regulation of AMOTL1 extends beyond cardiac fibroblasts is unknown\", \"Contribution relative to miRNA-mediated regulation not compared\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Biophysical reconstitution of AMOTL1's three PPxY motifs with NEDD4-1 and KIBRA WW domains revealed that cooperative multivalent engagement by NEDD4-1 (10-fold affinity enhancement) versus single-site binding by KIBRA explains how competing E3 ligase and protector complexes differentially regulate AMOTL1 stability.\",\n      \"evidence\": \"Isothermal titration calorimetry (ITC) and NMR spectroscopy with purified proteins\",\n      \"pmids\": [\"41580069\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular validation of cooperativity-dependent degradation rates not performed\", \"Whether post-translational modifications modulate WW–PPxY affinities in vivo is unknown\", \"KIBRA's protective effect not confirmed by in vivo genetic experiment\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Mapping of the Tankyrase-binding domain and identification of disease-associated R157C/P160L mutations that abolish TNKS1/2 and RNF146 binding, PARylation, and degradation established a direct PARylation-dependent degradation axis; stabilized mutants disrupt junctions and cause craniofacial and cardiac defects in zebrafish.\",\n      \"evidence\": \"Co-IP, PARylation and ubiquitination assays, live-cell migration imaging, zebrafish embryo expression\",\n      \"pmids\": [\"42012498\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Human genetic disease association not confirmed by family segregation or GWAS\", \"Whether R157C/P160L mutations affect NEDD4-1/KIBRA axis is untested\", \"Rescue of zebrafish phenotype by Tankyrase overexpression not attempted\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Discovery that PFKP directly binds and stabilizes AMOTL1 to promote YAP nuclear translocation linked glycolytic metabolism to Hippo pathway suppression via AMOTL1, explaining how metabolic reprogramming drives EMT in head and neck cancer.\",\n      \"evidence\": \"Co-IP, ubiquitination analysis, siRNA functional rescue, nude mouse xenograft\",\n      \"pmids\": [\"41727965\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which PFKP inhibits AMOTL1 ubiquitination is not defined\", \"Single-lab finding awaiting independent replication\", \"Whether PFKP enzymatic activity or scaffolding function is required is unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis of the AMOTL1–YAP1 interaction, the identity of the E3 ligase(s) targeting this complex, and whether AMOTL1 enters the nucleus as a co-factor or acts solely at the cytoplasmic-nuclear boundary remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No atomic-resolution structure of AMOTL1 or AMOTL1–YAP1 complex\", \"Nuclear versus cytoplasmic function of AMOTL1 not genetically separated\", \"Integration of multiple degradation axes (Tankyrase, NEDD4-1, Merlin, HECW2, PFKP, KIBRA) into a unified quantitative model not achieved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [5, 7, 12]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [7, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [7, 8, 10]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [5, 7, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [5, 7, 9, 12]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [0, 1, 3, 10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"YAP1\",\n      \"NEDD4\",\n      \"KIBRA\",\n      \"TNKS1\",\n      \"RNF146\",\n      \"HECW2\",\n      \"NF2\",\n      \"PFKP\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}