Affinage

AMOTL1

Angiomotin-like protein 1 · UniProt Q8IY63

Length
956 aa
Mass
106.6 kDa
Annotated
2026-04-28
21 papers in source corpus 14 papers cited in narrative 14 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

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).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 2001 Medium

    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

    PMID:11733531

    Open questions at the time
    • No functional perturbation performed
    • Mechanism of membrane recruitment not determined
    • No signaling pathway linkage established
  2. 2007 High

    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

    PMID:17397395

    Open questions at the time
    • No loss-of-function phenotype assessed
    • Other localization determinants not mapped
    • Functional consequence of MUPP1/Patj binding unknown
  3. 2016 High

    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)

    PMID:26806348 PMID:27498087

    Open questions at the time
    • 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
  4. 2017 High

    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

    PMID:28239148

    Open questions at the time
    • 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
  5. 2020 High

    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

    PMID:32313226

    Open questions at the time
    • 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
  6. 2023 Medium

    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

    PMID:37558679

    Open questions at the time
    • 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
  7. 2024 High

    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

    PMID:38839936

    Open questions at the time
    • 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
  8. 2026 High

    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

    PMID:41580069

    Open questions at the time
    • 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
  9. 2026 High

    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

    PMID:42012498

    Open questions at the time
    • 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
  10. 2026 Medium

    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

    PMID:41727965

    Open questions at the time
    • 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

Open questions

Synthesis pass · forward-looking unresolved questions
  • 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.
  • 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

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 3 GO:0098772 molecular function regulator activity 2
Localization
GO:0005634 nucleus 3 GO:0005829 cytosol 3 GO:0005886 plasma membrane 2
Pathway
R-HSA-1500931 Cell-Cell communication 4 R-HSA-162582 Signal Transduction 4
Partners
HECW2KIBRANEDD4NF2PFKPRNF146TNKS1YAP1

Evidence

Reading pass · 14 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2001 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. Fluorescence localization-based expression cloning, immunofluorescence microscopy, co-localization with TJ markers The Journal of biological chemistry Medium 11733531
2007 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. Yeast two-hybrid screening, immunofluorescence microscopy, biochemical fractionation, domain mapping Genes to cells : devoted to molecular & cellular mechanisms High 17397395
2014 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. 3'UTR luciferase reporter assay, miRNA overexpression, in vitro migration/invasion assays Cancer letters Medium 25218344
2016 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. Co-immunoprecipitation, ubiquitination assays (K63-linkage specific), siRNA knockdown, immunofluorescence, angiogenic sprouting assay Cellular signalling High 27498087
2016 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. Co-immunoprecipitation, proteasome inhibition assays, siRNA knockdown, cell migration assays, c-Src activity measurement Neoplasia (New York, N.Y.) Medium 26806348
2017 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. Mouse genetic knockout (Fat4 mutant), immunofluorescence for nuclear/cytoplasmic localization, cardiomyocyte proliferation assays, epistasis analysis Nature communications High 28239148
2019 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. qRT-PCR, gain/loss-of-function assays, luciferase reporter assay for ceRNA mechanism, in vivo xenograft Molecular therapy. Nucleic acids Medium 31812104
2020 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. Co-immunoprecipitation, immunofluorescence, ubiquitination assay, siRNA knockdown, xenograft assay Oncogene High 32313226
2023 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. Transcriptome analysis, RNA-protein binding assay (RRM domain mapping), immunofluorescence for localization, co-immunoprecipitation, functional rescue assays Cell death & disease Medium 37558679
2024 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. ac4C-RIP-seq, siRNA knockdown, fibroblast-specific Nat10 KO/OE mouse models, co-immunoprecipitation, verteporfin YAP inhibition Acta pharmacologica Sinica High 38839936
2026 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. Co-immunoprecipitation, ubiquitination assay, PARylation assay, live-cell imaging of migration, zebrafish embryo expression system Bioscience reports High 42012498
2026 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). Isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR) spectroscopy, quantitative molecular biophysical analyses Journal of molecular biology High 41580069
2026 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. Co-immunoprecipitation, ubiquitination analysis, immunofluorescence, siRNA knockdown functional rescue, in vivo nude mouse tumor model Journal of translational internal medicine Medium 41727965
2025 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. Tankyrase inhibitor treatment in primary lung fibroblasts, lung-on-a-chip, precision-cut lung slices, bleomycin mouse model; immunoblotting and qRT-PCR bioRxivpreprint Medium bio_10.1101_2025.11.13.688191

Source papers

Stage 0 corpus · 21 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 MiR-124 represses vasculogenic mimicry and cell motility by targeting amotL1 in cervical cancer cells. Cancer letters 83 25218344
2017 Amotl1 mediates sequestration of the Hippo effector Yap1 downstream of Fat4 to restrict heart growth. Nature communications 81 28239148
2007 Molecular characterization of angiomotin/JEAP family proteins: interaction with MUPP1/Patj and their endogenous properties. Genes to cells : devoted to molecular & cellular mechanisms 81 17397395
2019 circAMOTL1 Motivates AMOTL1 Expression to Facilitate Cervical Cancer Growth. Molecular therapy. Nucleic acids 73 31812104
2001 JEAP, a novel component of tight junctions in exocrine cells. The Journal of biological chemistry 58 11733531
2016 The endothelial E3 ligase HECW2 promotes endothelial cell junctions by increasing AMOTL1 protein stability via K63-linked ubiquitination. Cellular signalling 43 27498087
2020 AMOTL1 enhances YAP1 stability and promotes YAP1-driven gastric oncogenesis. Oncogene 37 32313226
2016 AMOTL1 Promotes Breast Cancer Progression and Is Antagonized by Merlin. Neoplasia (New York, N.Y.) 32 26806348
2020 The circ-AMOTL1/ENO1 Axis Implicated in the Tumorigenesis of OLP-Associated Oral Squamous Cell Carcinoma. Cancer management and research 17 32884340
2023 Circ-AMOTL1 enhances cardiac fibrosis through binding with EIF4A3 and stabilizing MARCKS expression in diabetic cardiomyopathy. Cellular signalling 13 37586467
2023 CircAMOTL1 RNA and AMOTL1 Protein: Complex Functions of AMOTL1 Gene Products. International journal of molecular sciences 8 36768425
2023 SRSF3/AMOTL1 splicing axis promotes the tumorigenesis of nasopharyngeal carcinoma through regulating the nucleus translocation of YAP1. Cell death & disease 7 37558679
2024 Acetylcytidine modification of Amotl1 by N-acetyltransferase 10 contributes to cardiac fibrotic expansion in mice after myocardial infarction. Acta pharmacologica Sinica 6 38839936
2024 circ-Amotl1 in extracellular vesicles derived from ADSCs improves wound healing by upregulating SPARC translation. Regenerative therapy 5 38318480
2023 A mutational hotspot in AMOTL1 defines a new syndrome of orofacial clefting, cardiac anomalies, and tall stature. American journal of medical genetics. Part A 4 36751037
2025 Hsa_circ_0001278 Facilitates Colorectal Cancer Progression via Sponging miR-338-5p and Regulating AMOTL1 Expression. Combinatorial chemistry & high throughput screening 3 38018210
2024 Harnessing machine learning and multi-omics to explore tumor evolutionary characteristics and the role of AMOTL1 in prostate cancer. International journal of biological macromolecules 2 39643184
2022 Clinical report: Chronic liver dysfunction in an individual with an AMOTL1 variant. European journal of medical genetics 1 36116699
2026 Multivalent AMOTL1 Engages NEDD4-1 and KIBRA Through Distinct Cooperative Binding Mechanisms. Journal of molecular biology 0 41580069
2026 PFKP binding AMOTL1 promotes tumor aerobic glycolysis and epithelial-mesenchymal transition by modulating Hippo pathway in head and neck cancer. Journal of translational internal medicine 0 41727965
2026 Patients-derived AMOTL1 mutations lead to defective cell migration and tissue development. Bioscience reports 0 42012498