{"gene":"AMER1","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2007,"finding":"WTX forms a complex with β-catenin, AXIN1, β-TrCP2, and APC (identified by tandem-affinity purification and mass spectrometry), and promotes β-catenin ubiquitination and degradation, thereby antagonizing WNT/β-catenin signaling in cultured cells, Xenopus, and zebrafish.","method":"Tandem-affinity protein purification, mass spectrometry, functional assays in cultured cells, Xenopus, and zebrafish","journal":"Science","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — TAP-MS complex identification combined with functional ubiquitination/degradation assays replicated across multiple model systems","pmids":["17510365"],"is_preprint":false},{"year":2007,"finding":"AMER1 (WTX) is a membrane-associated protein that recruits APC to the plasma membrane by binding to the ARM repeat domain of APC. The N-terminus of AMER1 contains two distinct PtdIns(4,5)P2-binding domains mediating plasma membrane localization. Overexpression redirects APC from microtubule ends to the plasma membrane; siRNA knockdown reduces APC levels, promotes microtubule-end association of APC, and disturbs intercellular junctions.","method":"Co-immunoprecipitation, siRNA knockdown, overexpression, live-cell imaging, domain mapping","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding assays combined with loss- and gain-of-function experiments with specific subcellular and morphological readouts, replicated in multiple cell lines","pmids":["17925383"],"is_preprint":false},{"year":2009,"finding":"WTX translocates to the nucleus (modulated by an endogenous splicing variant and a nuclear export inhibitor), co-localizes with the paraspeckle marker p54NRB/NONO, binds the WT1 zinc-finger transcription factor, and enhances WT1-mediated transcription of the endogenous target gene Amphiregulin.","method":"Nuclear fractionation, co-immunoprecipitation, co-localization/immunofluorescence, reporter transcription assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, nuclear localization confirmed by fractionation, functional transcription assay with endogenous target, single lab but multiple orthogonal methods","pmids":["19416806"],"is_preprint":false},{"year":2011,"finding":"Amer1/WTX directly interacts with the armadillo repeats of β-catenin via repeated REA (arginine-glutamic acid-alanine) motifs, assembles the β-catenin destruction complex (β-catenin, APC, Axin/Conductin) at the plasma membrane, stabilizes Axin, and counteracts Wnt-induced Axin degradation. Deletion or mutation of the membrane-binding domain abolishes membrane localization and abrogates negative Wnt regulation, which is restored by artificial membrane targeting.","method":"Co-immunoprecipitation, domain deletion/point mutagenesis, siRNA knockdown, Wnt target gene reporter assays, artificial membrane targeting","journal":"The Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct binding mapped by mutagenesis, multiple orthogonal functional assays including artificial rescue, consistent with other labs","pmids":["21498506"],"is_preprint":false},{"year":2011,"finding":"Amer1/WTX is essential for Wnt-induced LRP6 phosphorylation by GSK3β and CK1γ at the plasma membrane. Amer1 translocates to the plasma membrane in a PtdIns(4,5)P2-dependent manner after Wnt stimulation, binds CK1γ, recruits Axin and GSK3β to the membrane, and promotes Axin–LRP6 complex formation. Fusion of Amer1 to the cytoplasmic domain of LRP6 induces LRP6 phosphorylation and robust Wnt/β-catenin signaling.","method":"siRNA knockdown, overexpression, LRP6-Amer1 fusion construct, Co-immunoprecipitation, phosphorylation assays, PtdIns(4,5)P2 application","journal":"The EMBO Journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal functional assays (knockdown, fusion protein, PIP2 rescue), mechanistic epistasis established, single lab but rigorous","pmids":["21304492"],"is_preprint":false},{"year":2012,"finding":"WTX interacts with KEAP1, the ubiquitin ligase adaptor that regulates NRF2 ubiquitination. WTX and NRF2 compete for KEAP1 binding; unlike its role with β-catenin (where WTX promotes ubiquitination), WTX inhibits NRF2 ubiquitination and degradation, thereby protecting the antioxidant response. Loss of WTX leads to rapid NRF2 degradation and reduced cellular response to cytotoxic insult.","method":"Co-immunoprecipitation, competition binding assay, ubiquitination assay, WTX knockdown with NRF2 stability readout","journal":"The Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, competition assay, ubiquitination assay, and functional readout; multiple orthogonal methods in single lab","pmids":["22215675"],"is_preprint":false},{"year":2012,"finding":"The C-terminal domain of WTX binds to the DNA-binding domain of p53, enhances p53 binding to CBP, and increases CBP/p300-mediated acetylation of p53 at Lys 373/382. WTX knockdown accelerates CBP/p300 protein turnover and attenuates p53 acetylation. In p53-reconstitution experiments, depletion of WTX suppresses cell-cycle arrest, apoptosis, and p53 target-gene expression.","method":"Co-immunoprecipitation (domain mapping), p53 acetylation assay, WTX knockdown, p53 reconstitution, cell-cycle/apoptosis assays","journal":"Molecular Cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, PTM (acetylation) assay, functional rescue experiment; multiple methods in single lab","pmids":["22285752"],"is_preprint":false},{"year":2013,"finding":"β-arrestin promotes Wnt-induced LRP6 phosphorylation through a direct interaction with Amer1/WTX, regulating Amer1 membrane dynamics (measured by FRAP). β-arrestin is required for Wnt3a-induced Amer1 membrane translocation and interacts with PtdIns kinases PI4KIIα and PIP5KIβ; cells lacking β-arrestin cannot increase PtdIns(4,5)P2 levels in response to Wnt3a.","method":"Co-immunoprecipitation, FRAP (fluorescence recovery after photobleaching), LRP6 phosphorylation assay, β-arrestin knockout cells","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FRAP for membrane dynamics, Co-IP for interaction, functional assay; single lab with multiple methods","pmids":["24265322"],"is_preprint":false},{"year":2015,"finding":"Crystal structures of the APC-ARM domain in complex with Amer1 fragments A1, A2, and newly identified A4 revealed that all three, along with other APC-ARM binding partners (Asef, Sam68), use a common recognition pattern. Amer1-A3 binds the C-terminal side of APC-ARM through a bipartite mode. Composite mutations on either APC or Amer1 disrupting all four interfaces abrogated their association in cultured cells and impaired membrane recruitment of APC by Amer1.","method":"X-ray crystallography, GST pull-down, yeast two-hybrid, isothermal titration calorimetry (ITC), mutagenesis, co-immunoprecipitation in cultured cells","journal":"Cell Discovery","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structures with multiple binding sites determined, validated by ITC, pulldown, yeast two-hybrid, and cell-based mutagenesis; multiple orthogonal methods","pmids":["27462415"],"is_preprint":false},{"year":2011,"finding":"Wtx deletion in mice causes neonatal lethality, somatic overgrowth, and malformation of multiple mesenchyme-derived tissues (bone, fat, kidney, heart, spleen). In primary mesenchymal progenitor cells (MPCs), Wtx inactivation leads to aberrant β-catenin activation driving altered lineage fate decisions, while alternative pathways contribute to delayed differentiation of lineage-restricted cells.","method":"Conditional mouse knockout at different developmental stages, primary mesenchymal progenitor cell differentiation assays, β-catenin activation measurement","journal":"Developmental Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean in vivo KO with specific cellular phenotype, stage-specific inactivation, primary cell assays establishing pathway placement","pmids":["21571217"],"is_preprint":false},{"year":2015,"finding":"The transcriptional corepressor TRIM28 is the major binding partner of nuclear WTX. WTX interacts with the coiled-coil domain of TRIM28 via its own coiled-coil and proline-rich domains. WTX knockdown reduces TRIM28 recruitment to a chromatinized reporter and impairs repression of target transcripts including endogenous retroviruses and LINEs. In mesenchymal precursor cells, co-depletion of WTX and TRIM28 causes β-catenin-independent defects in adipogenic and osteogenic differentiation, and WTX knockdown reduces TRIM28 binding to the Pparγ promoter.","method":"Co-immunoprecipitation (domain mapping), chromatin immunoprecipitation (ChIP), single-molecule RNA sequencing, siRNA knockdown, differentiation assays","journal":"The Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, ChIP for chromatin recruitment, smRNA-seq for transcriptional output, differentiation phenotype; multiple orthogonal methods single lab","pmids":["25882849"],"is_preprint":false},{"year":2010,"finding":"Functional analysis of WTX and tumor-associated truncation mutants shows that WTX induction causes G1/G0 cell-cycle arrest and growth inhibition through p21 induction. The short truncation mutant WTX358 lacks this activity, while the longer truncation WTX565 retains it, mapping the critical functional region to amino acids 358–561. Gene expression profiling showed overlap between WTX and WT1-regulated pathways.","method":"Inducible cell-line expression, cell-cycle analysis (flow cytometry), gene expression profiling, truncation mutant analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — inducible expression system, cell-cycle assay, gene profiling, domain mapping by truncation; single lab, multiple methods","pmids":["20956941"],"is_preprint":false},{"year":2020,"finding":"KIF23 directly interacts with Amer1 and competitively blocks the Amer1–APC interaction, thereby relocating Amer1 from the membrane/cytoplasm to the nucleus and attenuating Amer1-mediated negative regulation of Wnt/β-catenin signaling, resulting in pathway activation in gastric cancer cells.","method":"Co-immunoprecipitation, competitive binding assay, subcellular fractionation/immunofluorescence, Wnt reporter assay, KIF23 knockdown/overexpression","journal":"Aging","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP for direct interaction, competitive binding, subcellular relocalization assay, functional Wnt reporter; single lab","pmids":["32365332"],"is_preprint":false},{"year":2019,"finding":"WTX loss in colorectal cancer disrupts the interaction between RhoGDIα and CDC42, triggering CDC42 activation and its downstream cascades to promote tumor invasion and metastasis. WTX binds RhoGDIα; loss of WTX prevents RhoGDIα from suppressing CDC42 activity.","method":"Co-immunoprecipitation, CDC42 activity assay, WTX knockdown/overexpression, in vivo metastasis model","journal":"Nature Communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating WTX-RhoGDIα binding, GTPase activity assay, in vivo validation; single lab but multiple methods","pmids":["30631060"],"is_preprint":false},{"year":2025,"finding":"The long isoform of WTX (WTX-L) promotes ferroptosis in gastric cancer by competitively interacting with β-arrestin2, disrupting its binding to IκBα and thereby activating the NF-κB/LCN2 pathway. LCN2 increases the labile Fe2+ pool and promotes lipid peroxidation to trigger ferroptosis.","method":"Co-immunoprecipitation (competitive), WTX-L knockdown/overexpression, NF-κB reporter, ferroptosis inducers (erastin/RSL3), in vivo xenograft","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — competitive Co-IP, pathway reporter, and in vivo functional assay; single lab, multiple methods","pmids":["40109379"],"is_preprint":false},{"year":2026,"finding":"The short isoform WTX-S interacts with NONO via its coiled-coil domain 2 binding to the NONO coiled-coil domain, driving formation of nuclear bodies (WTX-NB) through liquid-liquid phase separation. This sequesters NONO from paraspeckles, disrupting paraspeckle stability and enhancing chemotherapy sensitivity in gastric cancer.","method":"Co-immunoprecipitation (domain mapping), liquid-liquid phase separation assay, live-cell imaging, siRNA knockdown, chemosensitivity assay","journal":"Cancer Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with domain mapping, LLPS assay, nuclear body imaging, functional chemosensitivity readout; single lab, multiple orthogonal methods","pmids":["41115290"],"is_preprint":false},{"year":2018,"finding":"A C-terminal truncation of WTX/AMER1 (modeled in mice) fails to co-immunoprecipitate β-catenin directly, but partial recruitment of β-catenin can occur indirectly via AXIN/AXIN2 as a molecular bridge, explaining how some truncation mutations retain partial WTX function.","method":"Co-immunoprecipitation in vitro, mouse knock-in allele generation, β-catenin recruitment assay","journal":"Journal of Bone and Mineral Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with truncation mutants in a knock-in mouse model, indirect recruitment via Axin demonstrated; single lab","pmids":["29329488"],"is_preprint":false},{"year":2022,"finding":"IRF-2 directly activates AMER1 transcription (established by ChIP-seq), thereby regulating the Wnt/β-catenin signaling pathway and inhibiting gastric cancer cell proliferation.","method":"Chromatin immunoprecipitation sequencing (ChIP-Seq), western blot, cell proliferation assays, xenograft model","journal":"Journal of Translational Medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP-Seq directly showing IRF-2 binding to AMER1 locus, functional proliferation assay; single lab","pmids":["35115027"],"is_preprint":false},{"year":2008,"finding":"Germline loss-of-function mutations in WTX cause osteopathia striata congenita with cranial sclerosis (OSCS), an X-linked sclerosing bone dysplasia, with the mouse homolog expressed in fetal skeleton. Alternative splicing implicating plasma membrane localization of WTX is associated with male survival in OSCS, linking subcellular localization to functional outcome.","method":"Germline mutation identification, mouse expression analysis, alternative splicing analysis, genotype-phenotype correlation","journal":"Nature Genetics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — human germline mutation-phenotype linkage across multiple families combined with mouse expression data; splicing/localization inference is indirect","pmids":["19079258"],"is_preprint":false},{"year":2019,"finding":"In zebrafish, wtx knockdown increases Wnt activity and causes embryonic dorsalization. In wtx mutants, Wnt target genes are transiently upregulated during caudal fin regeneration, confirming a negative regulatory role in Wnt signaling in vivo. However, wtx single or wtx/amer2/amer3 triple mutants develop normally and are fertile, demonstrating that the zebrafish Wtx/Amer family members do not compensate for each other.","method":"Morpholino knockdown, TALEN knockout, Wnt target gene expression analysis, fin regeneration assay","journal":"Developmental Dynamics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — morpholino and TALEN knockout with Wnt reporter readout; single lab, orthologous zebrafish model","pmids":["31290212"],"is_preprint":false}],"current_model":"AMER1/WTX is a multifunctional scaffold protein that acts as a context-dependent regulator of Wnt/β-catenin signaling: at the plasma membrane it assembles the β-catenin destruction complex (with APC, Axin, β-TrCP2) by binding PtdIns(4,5)P2 and directly interacts with the APC-ARM domain and β-catenin REA motifs to promote β-catenin ubiquitination/degradation, while also facilitating LRP6 phosphorylation by recruiting CK1γ and GSK3β; in the nucleus it binds WT1 to enhance target-gene transcription, associates with TRIM28 for epigenetic silencing and regulation of differentiation, interacts with p53 to promote CBP/p300-mediated p53 acetylation, and via its short isoform drives NONO sequestration and paraspeckle disintegration through liquid-liquid phase separation; additionally, WTX competes with NRF2 for KEAP1 binding to inhibit NRF2 ubiquitination, interacts with RhoGDIα to suppress CDC42 activation, and through its long isoform engages β-arrestin2 to activate NF-κB/LCN2-dependent ferroptosis."},"narrative":{"mechanistic_narrative":"AMER1 (WTX) is a membrane-associated scaffold protein that functions as a negative regulator of Wnt/β-catenin signaling, assembling the β-catenin destruction complex at the plasma membrane and promoting β-catenin ubiquitination and degradation [PMID:17510365]. It localizes to the membrane through two N-terminal PtdIns(4,5)P2-binding domains and recruits APC there by binding the APC armadillo-repeat domain through multiple interfaces resolved by crystallography [PMID:17925383, PMID:27462415]. AMER1 directly engages β-catenin armadillo repeats via tandem REA motifs, stabilizes Axin, and counteracts Wnt-induced Axin degradation; tumor-associated C-terminal truncations lose direct β-catenin binding but retain partial activity through Axin-bridged recruitment [PMID:21498506, PMID:29329488]. Paradoxically, AMER1 is also required for Wnt-induced LRP6 phosphorylation, translocating to the membrane in a PtdIns(4,5)P2-dependent manner to recruit CK1γ, Axin and GSK3β and promote Axin–LRP6 complex formation, with β-arrestin governing its membrane dynamics and PtdIns(4,5)P2 production [PMID:21304492, PMID:24265322]. In vivo, Wtx loss causes neonatal lethality, somatic overgrowth, and aberrant β-catenin–driven mesenchymal lineage decisions, and germline loss-of-function mutations cause the X-linked sclerosing bone dysplasia osteopathia striata congenita with cranial sclerosis [PMID:21571217, PMID:19079258]. Beyond Wnt, AMER1 acts in the nucleus, binding the WT1 transcription factor to enhance target-gene transcription, partnering with TRIM28 to repress endogenous retroviruses and LINEs and to control adipogenic/osteogenic differentiation, and binding p53 to promote CBP/p300-mediated acetylation and p53-dependent cell-cycle arrest and apoptosis [PMID:19416806, PMID:25882849, PMID:22285752]. It additionally competes with NRF2 for KEAP1 binding to stabilize NRF2 and protect the antioxidant response, and binds RhoGDIα to restrain CDC42 activation [PMID:22215675, PMID:30631060]. Isoform-specific functions include WTX-S driving NONO sequestration and paraspeckle disassembly via liquid-liquid phase separation, and WTX-L promoting NF-κB/LCN2-dependent ferroptosis through competitive binding to β-arrestin2 [PMID:41115290, PMID:40109379].","teleology":[{"year":2007,"claim":"Established AMER1/WTX as a bona fide component of the β-catenin destruction complex, defining its core role as a Wnt antagonist that promotes β-catenin turnover.","evidence":"Tandem-affinity purification/mass spectrometry and ubiquitination/degradation assays in cells, Xenopus, and zebrafish","pmids":["17510365"],"confidence":"High","gaps":["Did not resolve where in the cell the complex assembles","Mechanism of how WTX promotes ubiquitination not detailed"]},{"year":2007,"claim":"Defined WTX as a membrane-anchored protein that recruits APC to the plasma membrane, placing destruction-complex assembly at the membrane.","evidence":"Co-IP, domain mapping of two PtdIns(4,5)P2-binding domains, siRNA, overexpression, and live imaging in multiple cell lines","pmids":["17925383"],"confidence":"High","gaps":["Structural basis of APC-ARM binding not yet determined","Functional consequence of membrane vs microtubule APC pools unresolved"]},{"year":2010,"claim":"Linked WTX expression to growth suppression via p21-mediated G1/G0 arrest and mapped the responsible region, framing tumor-associated truncations as functional losses.","evidence":"Inducible expression, flow-cytometry cell-cycle analysis, truncation mutant mapping, and expression profiling","pmids":["20956941"],"confidence":"Medium","gaps":["Molecular link between WTX and p21 induction not established","Single inducible cell system"]},{"year":2011,"claim":"Showed WTX directly binds β-catenin armadillo repeats through REA motifs and stabilizes Axin at the membrane, defining the molecular interface for destruction-complex assembly.","evidence":"Co-IP, domain deletion/point mutagenesis, reporter assays, and artificial membrane-targeting rescue","pmids":["21498506"],"confidence":"High","gaps":["Stoichiometry of the membrane destruction complex unresolved","How membrane assembly couples to ubiquitination not detailed"]},{"year":2011,"claim":"Revealed the paradoxical positive role of WTX in Wnt-induced LRP6 phosphorylation, recruiting CK1γ/GSK3β/Axin to the membrane and producing the receptor-activating signal.","evidence":"siRNA, LRP6-Amer1 fusion construct, phosphorylation assays, and PtdIns(4,5)P2 application","pmids":["21304492"],"confidence":"High","gaps":["How the same protein both activates LRP6 and degrades β-catenin is not mechanistically reconciled","Temporal switch between roles unknown"]},{"year":2011,"claim":"Demonstrated in vivo that Wtx is essential for development and that its loss drives β-catenin–dependent mesenchymal lineage defects.","evidence":"Conditional/stage-specific mouse knockout and primary mesenchymal progenitor differentiation assays","pmids":["21571217"],"confidence":"High","gaps":["β-catenin-independent contributions to differentiation delay not molecularly defined"]},{"year":2009,"claim":"Identified a nuclear function for WTX as a WT1 transcriptional cofactor and noted association with the paraspeckle marker NONO.","evidence":"Nuclear fractionation, Co-IP, co-localization, and WT1 target reporter assays","pmids":["19416806"],"confidence":"High","gaps":["Functional significance of NONO co-localization not yet established","Regulation of nuclear vs membrane partitioning incompletely defined"]},{"year":2012,"claim":"Showed WTX inhibits rather than promotes ubiquitination in the KEAP1–NRF2 axis, competing with NRF2 to stabilize the antioxidant response and revealing context-dependent E3-adaptor effects.","evidence":"Co-IP, competition binding, ubiquitination assay, and NRF2 stability readout on WTX knockdown","pmids":["22215675"],"confidence":"High","gaps":["Why WTX promotes β-catenin but blocks NRF2 ubiquitination not mechanistically explained"]},{"year":2012,"claim":"Established WTX as a p53 cofactor that enhances CBP/p300-mediated p53 acetylation and p53-dependent arrest/apoptosis, broadening its tumor-suppressor functions.","evidence":"Domain-mapping Co-IP, p53 acetylation assays, knockdown, and p53 reconstitution with cell-cycle/apoptosis readouts","pmids":["22285752"],"confidence":"High","gaps":["How WTX stabilizes CBP/p300 not defined","Crosstalk with WTX Wnt functions unexplored"]},{"year":2013,"claim":"Identified β-arrestin as a regulator of WTX membrane dynamics and PtdIns(4,5)P2 production during Wnt-induced LRP6 phosphorylation.","evidence":"Co-IP, FRAP, LRP6 phosphorylation assays in β-arrestin knockout cells","pmids":["24265322"],"confidence":"Medium","gaps":["Direct vs indirect coupling to PI kinases not fully resolved","Single-lab characterization"]},{"year":2015,"claim":"Determined the structural basis of WTX–APC recognition, showing multiple Amer1 fragments bind APC-ARM using a shared mode shared with other partners.","evidence":"X-ray crystallography, ITC, GST pull-down, yeast two-hybrid, and cell-based composite mutagenesis","pmids":["27462415"],"confidence":"High","gaps":["Structure of WTX with β-catenin or Axin not solved","Full-length WTX architecture unknown"]},{"year":2015,"claim":"Defined TRIM28 as the major nuclear partner of WTX, establishing a β-catenin-independent role in epigenetic silencing of retroelements and control of mesenchymal differentiation.","evidence":"Domain-mapping Co-IP, ChIP, single-molecule RNA-seq, and differentiation assays","pmids":["25882849"],"confidence":"High","gaps":["Whether WTX directly contacts chromatin unresolved","How WTX is targeted to specific loci unknown"]},{"year":2018,"claim":"Explained the partial function of C-terminal WTX truncations by showing Axin can indirectly bridge β-catenin recruitment when direct binding is lost.","evidence":"In vitro Co-IP with truncation mutants and a knock-in mouse allele","pmids":["29329488"],"confidence":"Medium","gaps":["Quantitative contribution of indirect bridging in vivo not measured"]},{"year":2019,"claim":"Extended WTX's negative Wnt role to vertebrate development and regeneration and showed Amer family members do not functionally compensate.","evidence":"Zebrafish morpholino knockdown, TALEN knockout, Wnt target analysis, and fin regeneration assay","pmids":["31290212"],"confidence":"Medium","gaps":["Normal development of mutants despite Wnt dysregulation not reconciled with mouse lethality"]},{"year":2019,"claim":"Uncovered a Wnt-independent cytoskeletal/invasion function in which WTX supports RhoGDIα suppression of CDC42, with loss driving metastasis.","evidence":"Co-IP, CDC42 activity assays, knockdown/overexpression, and in vivo metastasis model","pmids":["30631060"],"confidence":"Medium","gaps":["Direct vs indirect WTX–RhoGDIα binding not structurally defined","Single-lab finding"]},{"year":2020,"claim":"Identified KIF23 as a competitive disruptor of the Amer1–APC interaction that relocalizes Amer1 to the nucleus and activates Wnt signaling in cancer.","evidence":"Co-IP, competitive binding, subcellular relocalization, and Wnt reporter assays","pmids":["32365332"],"confidence":"Medium","gaps":["Whether nuclear-relocalized Amer1 gains nuclear function not tested","Single-lab finding"]},{"year":2022,"claim":"Placed AMER1 downstream of IRF-2 transcriptional control, identifying an upstream regulator of its expression that constrains Wnt-driven proliferation.","evidence":"ChIP-seq, western blot, proliferation assays, and xenograft model","pmids":["35115027"],"confidence":"Medium","gaps":["Other transcriptional regulators of AMER1 not surveyed"]},{"year":2025,"claim":"Defined an isoform-specific WTX-L function promoting ferroptosis via competitive β-arrestin2 binding and NF-κB/LCN2 activation.","evidence":"Competitive Co-IP, NF-κB reporter, ferroptosis inducers, and xenograft","pmids":["40109379"],"confidence":"Medium","gaps":["Generality beyond gastric cancer unknown","How isoform choice is regulated unclear"]},{"year":2026,"claim":"Defined an isoform-specific WTX-S function that sequesters NONO into phase-separated nuclear bodies to disassemble paraspeckles and sensitize cells to chemotherapy.","evidence":"Domain-mapping Co-IP, LLPS assay, live-cell imaging, and chemosensitivity assays","pmids":["41115290"],"confidence":"Medium","gaps":["Physiological triggers of WTX-S nuclear body formation unknown","Single-lab finding"]},{"year":null,"claim":"How WTX integrates its opposing roles — destruction-complex scaffold versus LRP6-activating cofactor, and pro- versus anti-ubiquitination E3-adaptor functions — within a single regulatory logic remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model reconciles WTX's positive and negative Wnt roles","Determinants of isoform- and partner-specific function not defined","Full-length structure and complex stoichiometry unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,3,4,8]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[1,4]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[5,6,13]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[2,10,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,3,4]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,10,6]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,12]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,3,4,9]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[2,10,6]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[5]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[9,18]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[14,6]}],"complexes":["β-catenin destruction complex"],"partners":["APC","CTNNB1","AXIN1","KEAP1","TP53","TRIM28","NONO","ARRB2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q5JTC6","full_name":"APC membrane recruitment protein 1","aliases":["Protein FAM123B","Wilms tumor gene on the X chromosome protein"],"length_aa":1135,"mass_kda":124.0,"function":"Regulator of the canonical Wnt signaling pathway. Acts by specifically binding phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), translocating to the cell membrane and interacting with key regulators of the canonical Wnt signaling pathway, such as components of the beta-catenin destruction complex. Acts both as a positive and negative regulator of the Wnt signaling pathway, depending on the context: acts as a positive regulator by promoting LRP6 phosphorylation. Also acts as a negative regulator by acting as a scaffold protein for the beta-catenin destruction complex and promoting stabilization of Axin at the cell membrane. Promotes CTNNB1 ubiquitination and degradation. Involved in kidney development","subcellular_location":"Cytoplasm; Cell membrane; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q5JTC6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMER1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AMER1","total_profiled":1310},"omim":[{"mim_id":"619975","title":"TUMOR PREDISPOSITION SYNDROME 2; TPDS2","url":"https://www.omim.org/entry/619975"},{"mim_id":"614659","title":"APC MEMBRANE RECRUITMENT PROTEIN 2; AMER2","url":"https://www.omim.org/entry/614659"},{"mim_id":"603574","title":"METHYL-CpG-BINDING DOMAIN PROTEIN 4; MBD4","url":"https://www.omim.org/entry/603574"},{"mim_id":"300647","title":"APC MEMBRANE RECRUITMENT PROTEIN 1; 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The N-terminus of AMER1 contains two distinct PtdIns(4,5)P2-binding domains mediating plasma membrane localization. Overexpression redirects APC from microtubule ends to the plasma membrane; siRNA knockdown reduces APC levels, promotes microtubule-end association of APC, and disturbs intercellular junctions.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, overexpression, live-cell imaging, domain mapping\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding assays combined with loss- and gain-of-function experiments with specific subcellular and morphological readouts, replicated in multiple cell lines\",\n      \"pmids\": [\"17925383\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"WTX translocates to the nucleus (modulated by an endogenous splicing variant and a nuclear export inhibitor), co-localizes with the paraspeckle marker p54NRB/NONO, binds the WT1 zinc-finger transcription factor, and enhances WT1-mediated transcription of the endogenous target gene Amphiregulin.\",\n      \"method\": \"Nuclear fractionation, co-immunoprecipitation, co-localization/immunofluorescence, reporter transcription assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, nuclear localization confirmed by fractionation, functional transcription assay with endogenous target, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"19416806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Amer1/WTX directly interacts with the armadillo repeats of β-catenin via repeated REA (arginine-glutamic acid-alanine) motifs, assembles the β-catenin destruction complex (β-catenin, APC, Axin/Conductin) at the plasma membrane, stabilizes Axin, and counteracts Wnt-induced Axin degradation. Deletion or mutation of the membrane-binding domain abolishes membrane localization and abrogates negative Wnt regulation, which is restored by artificial membrane targeting.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion/point mutagenesis, siRNA knockdown, Wnt target gene reporter assays, artificial membrane targeting\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct binding mapped by mutagenesis, multiple orthogonal functional assays including artificial rescue, consistent with other labs\",\n      \"pmids\": [\"21498506\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Amer1/WTX is essential for Wnt-induced LRP6 phosphorylation by GSK3β and CK1γ at the plasma membrane. Amer1 translocates to the plasma membrane in a PtdIns(4,5)P2-dependent manner after Wnt stimulation, binds CK1γ, recruits Axin and GSK3β to the membrane, and promotes Axin–LRP6 complex formation. Fusion of Amer1 to the cytoplasmic domain of LRP6 induces LRP6 phosphorylation and robust Wnt/β-catenin signaling.\",\n      \"method\": \"siRNA knockdown, overexpression, LRP6-Amer1 fusion construct, Co-immunoprecipitation, phosphorylation assays, PtdIns(4,5)P2 application\",\n      \"journal\": \"The EMBO Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal functional assays (knockdown, fusion protein, PIP2 rescue), mechanistic epistasis established, single lab but rigorous\",\n      \"pmids\": [\"21304492\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"WTX interacts with KEAP1, the ubiquitin ligase adaptor that regulates NRF2 ubiquitination. WTX and NRF2 compete for KEAP1 binding; unlike its role with β-catenin (where WTX promotes ubiquitination), WTX inhibits NRF2 ubiquitination and degradation, thereby protecting the antioxidant response. Loss of WTX leads to rapid NRF2 degradation and reduced cellular response to cytotoxic insult.\",\n      \"method\": \"Co-immunoprecipitation, competition binding assay, ubiquitination assay, WTX knockdown with NRF2 stability readout\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, competition assay, ubiquitination assay, and functional readout; multiple orthogonal methods in single lab\",\n      \"pmids\": [\"22215675\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The C-terminal domain of WTX binds to the DNA-binding domain of p53, enhances p53 binding to CBP, and increases CBP/p300-mediated acetylation of p53 at Lys 373/382. WTX knockdown accelerates CBP/p300 protein turnover and attenuates p53 acetylation. In p53-reconstitution experiments, depletion of WTX suppresses cell-cycle arrest, apoptosis, and p53 target-gene expression.\",\n      \"method\": \"Co-immunoprecipitation (domain mapping), p53 acetylation assay, WTX knockdown, p53 reconstitution, cell-cycle/apoptosis assays\",\n      \"journal\": \"Molecular Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, PTM (acetylation) assay, functional rescue experiment; multiple methods in single lab\",\n      \"pmids\": [\"22285752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"β-arrestin promotes Wnt-induced LRP6 phosphorylation through a direct interaction with Amer1/WTX, regulating Amer1 membrane dynamics (measured by FRAP). β-arrestin is required for Wnt3a-induced Amer1 membrane translocation and interacts with PtdIns kinases PI4KIIα and PIP5KIβ; cells lacking β-arrestin cannot increase PtdIns(4,5)P2 levels in response to Wnt3a.\",\n      \"method\": \"Co-immunoprecipitation, FRAP (fluorescence recovery after photobleaching), LRP6 phosphorylation assay, β-arrestin knockout cells\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FRAP for membrane dynamics, Co-IP for interaction, functional assay; single lab with multiple methods\",\n      \"pmids\": [\"24265322\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Crystal structures of the APC-ARM domain in complex with Amer1 fragments A1, A2, and newly identified A4 revealed that all three, along with other APC-ARM binding partners (Asef, Sam68), use a common recognition pattern. Amer1-A3 binds the C-terminal side of APC-ARM through a bipartite mode. Composite mutations on either APC or Amer1 disrupting all four interfaces abrogated their association in cultured cells and impaired membrane recruitment of APC by Amer1.\",\n      \"method\": \"X-ray crystallography, GST pull-down, yeast two-hybrid, isothermal titration calorimetry (ITC), mutagenesis, co-immunoprecipitation in cultured cells\",\n      \"journal\": \"Cell Discovery\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structures with multiple binding sites determined, validated by ITC, pulldown, yeast two-hybrid, and cell-based mutagenesis; multiple orthogonal methods\",\n      \"pmids\": [\"27462415\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Wtx deletion in mice causes neonatal lethality, somatic overgrowth, and malformation of multiple mesenchyme-derived tissues (bone, fat, kidney, heart, spleen). In primary mesenchymal progenitor cells (MPCs), Wtx inactivation leads to aberrant β-catenin activation driving altered lineage fate decisions, while alternative pathways contribute to delayed differentiation of lineage-restricted cells.\",\n      \"method\": \"Conditional mouse knockout at different developmental stages, primary mesenchymal progenitor cell differentiation assays, β-catenin activation measurement\",\n      \"journal\": \"Developmental Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean in vivo KO with specific cellular phenotype, stage-specific inactivation, primary cell assays establishing pathway placement\",\n      \"pmids\": [\"21571217\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The transcriptional corepressor TRIM28 is the major binding partner of nuclear WTX. WTX interacts with the coiled-coil domain of TRIM28 via its own coiled-coil and proline-rich domains. WTX knockdown reduces TRIM28 recruitment to a chromatinized reporter and impairs repression of target transcripts including endogenous retroviruses and LINEs. In mesenchymal precursor cells, co-depletion of WTX and TRIM28 causes β-catenin-independent defects in adipogenic and osteogenic differentiation, and WTX knockdown reduces TRIM28 binding to the Pparγ promoter.\",\n      \"method\": \"Co-immunoprecipitation (domain mapping), chromatin immunoprecipitation (ChIP), single-molecule RNA sequencing, siRNA knockdown, differentiation assays\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, ChIP for chromatin recruitment, smRNA-seq for transcriptional output, differentiation phenotype; multiple orthogonal methods single lab\",\n      \"pmids\": [\"25882849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Functional analysis of WTX and tumor-associated truncation mutants shows that WTX induction causes G1/G0 cell-cycle arrest and growth inhibition through p21 induction. The short truncation mutant WTX358 lacks this activity, while the longer truncation WTX565 retains it, mapping the critical functional region to amino acids 358–561. Gene expression profiling showed overlap between WTX and WT1-regulated pathways.\",\n      \"method\": \"Inducible cell-line expression, cell-cycle analysis (flow cytometry), gene expression profiling, truncation mutant analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — inducible expression system, cell-cycle assay, gene profiling, domain mapping by truncation; single lab, multiple methods\",\n      \"pmids\": [\"20956941\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"KIF23 directly interacts with Amer1 and competitively blocks the Amer1–APC interaction, thereby relocating Amer1 from the membrane/cytoplasm to the nucleus and attenuating Amer1-mediated negative regulation of Wnt/β-catenin signaling, resulting in pathway activation in gastric cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, competitive binding assay, subcellular fractionation/immunofluorescence, Wnt reporter assay, KIF23 knockdown/overexpression\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP for direct interaction, competitive binding, subcellular relocalization assay, functional Wnt reporter; single lab\",\n      \"pmids\": [\"32365332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"WTX loss in colorectal cancer disrupts the interaction between RhoGDIα and CDC42, triggering CDC42 activation and its downstream cascades to promote tumor invasion and metastasis. WTX binds RhoGDIα; loss of WTX prevents RhoGDIα from suppressing CDC42 activity.\",\n      \"method\": \"Co-immunoprecipitation, CDC42 activity assay, WTX knockdown/overexpression, in vivo metastasis model\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating WTX-RhoGDIα binding, GTPase activity assay, in vivo validation; single lab but multiple methods\",\n      \"pmids\": [\"30631060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The long isoform of WTX (WTX-L) promotes ferroptosis in gastric cancer by competitively interacting with β-arrestin2, disrupting its binding to IκBα and thereby activating the NF-κB/LCN2 pathway. LCN2 increases the labile Fe2+ pool and promotes lipid peroxidation to trigger ferroptosis.\",\n      \"method\": \"Co-immunoprecipitation (competitive), WTX-L knockdown/overexpression, NF-κB reporter, ferroptosis inducers (erastin/RSL3), in vivo xenograft\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — competitive Co-IP, pathway reporter, and in vivo functional assay; single lab, multiple methods\",\n      \"pmids\": [\"40109379\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The short isoform WTX-S interacts with NONO via its coiled-coil domain 2 binding to the NONO coiled-coil domain, driving formation of nuclear bodies (WTX-NB) through liquid-liquid phase separation. This sequesters NONO from paraspeckles, disrupting paraspeckle stability and enhancing chemotherapy sensitivity in gastric cancer.\",\n      \"method\": \"Co-immunoprecipitation (domain mapping), liquid-liquid phase separation assay, live-cell imaging, siRNA knockdown, chemosensitivity assay\",\n      \"journal\": \"Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with domain mapping, LLPS assay, nuclear body imaging, functional chemosensitivity readout; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"41115290\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A C-terminal truncation of WTX/AMER1 (modeled in mice) fails to co-immunoprecipitate β-catenin directly, but partial recruitment of β-catenin can occur indirectly via AXIN/AXIN2 as a molecular bridge, explaining how some truncation mutations retain partial WTX function.\",\n      \"method\": \"Co-immunoprecipitation in vitro, mouse knock-in allele generation, β-catenin recruitment assay\",\n      \"journal\": \"Journal of Bone and Mineral Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with truncation mutants in a knock-in mouse model, indirect recruitment via Axin demonstrated; single lab\",\n      \"pmids\": [\"29329488\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"IRF-2 directly activates AMER1 transcription (established by ChIP-seq), thereby regulating the Wnt/β-catenin signaling pathway and inhibiting gastric cancer cell proliferation.\",\n      \"method\": \"Chromatin immunoprecipitation sequencing (ChIP-Seq), western blot, cell proliferation assays, xenograft model\",\n      \"journal\": \"Journal of Translational Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP-Seq directly showing IRF-2 binding to AMER1 locus, functional proliferation assay; single lab\",\n      \"pmids\": [\"35115027\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Germline loss-of-function mutations in WTX cause osteopathia striata congenita with cranial sclerosis (OSCS), an X-linked sclerosing bone dysplasia, with the mouse homolog expressed in fetal skeleton. Alternative splicing implicating plasma membrane localization of WTX is associated with male survival in OSCS, linking subcellular localization to functional outcome.\",\n      \"method\": \"Germline mutation identification, mouse expression analysis, alternative splicing analysis, genotype-phenotype correlation\",\n      \"journal\": \"Nature Genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — human germline mutation-phenotype linkage across multiple families combined with mouse expression data; splicing/localization inference is indirect\",\n      \"pmids\": [\"19079258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In zebrafish, wtx knockdown increases Wnt activity and causes embryonic dorsalization. In wtx mutants, Wnt target genes are transiently upregulated during caudal fin regeneration, confirming a negative regulatory role in Wnt signaling in vivo. However, wtx single or wtx/amer2/amer3 triple mutants develop normally and are fertile, demonstrating that the zebrafish Wtx/Amer family members do not compensate for each other.\",\n      \"method\": \"Morpholino knockdown, TALEN knockout, Wnt target gene expression analysis, fin regeneration assay\",\n      \"journal\": \"Developmental Dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — morpholino and TALEN knockout with Wnt reporter readout; single lab, orthologous zebrafish model\",\n      \"pmids\": [\"31290212\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AMER1/WTX is a multifunctional scaffold protein that acts as a context-dependent regulator of Wnt/β-catenin signaling: at the plasma membrane it assembles the β-catenin destruction complex (with APC, Axin, β-TrCP2) by binding PtdIns(4,5)P2 and directly interacts with the APC-ARM domain and β-catenin REA motifs to promote β-catenin ubiquitination/degradation, while also facilitating LRP6 phosphorylation by recruiting CK1γ and GSK3β; in the nucleus it binds WT1 to enhance target-gene transcription, associates with TRIM28 for epigenetic silencing and regulation of differentiation, interacts with p53 to promote CBP/p300-mediated p53 acetylation, and via its short isoform drives NONO sequestration and paraspeckle disintegration through liquid-liquid phase separation; additionally, WTX competes with NRF2 for KEAP1 binding to inhibit NRF2 ubiquitination, interacts with RhoGDIα to suppress CDC42 activation, and through its long isoform engages β-arrestin2 to activate NF-κB/LCN2-dependent ferroptosis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AMER1 (WTX) is a membrane-associated scaffold protein that functions as a negative regulator of Wnt/\\u03b2-catenin signaling, assembling the \\u03b2-catenin destruction complex at the plasma membrane and promoting \\u03b2-catenin ubiquitination and degradation [#0]. It localizes to the membrane through two N-terminal PtdIns(4,5)P2-binding domains and recruits APC there by binding the APC armadillo-repeat domain through multiple interfaces resolved by crystallography [#1, #8]. AMER1 directly engages \\u03b2-catenin armadillo repeats via tandem REA motifs, stabilizes Axin, and counteracts Wnt-induced Axin degradation; tumor-associated C-terminal truncations lose direct \\u03b2-catenin binding but retain partial activity through Axin-bridged recruitment [#3, #16]. Paradoxically, AMER1 is also required for Wnt-induced LRP6 phosphorylation, translocating to the membrane in a PtdIns(4,5)P2-dependent manner to recruit CK1\\u03b3, Axin and GSK3\\u03b2 and promote Axin\\u2013LRP6 complex formation, with \\u03b2-arrestin governing its membrane dynamics and PtdIns(4,5)P2 production [#4, #7]. In vivo, Wtx loss causes neonatal lethality, somatic overgrowth, and aberrant \\u03b2-catenin\\u2013driven mesenchymal lineage decisions, and germline loss-of-function mutations cause the X-linked sclerosing bone dysplasia osteopathia striata congenita with cranial sclerosis [#9, #18]. Beyond Wnt, AMER1 acts in the nucleus, binding the WT1 transcription factor to enhance target-gene transcription, partnering with TRIM28 to repress endogenous retroviruses and LINEs and to control adipogenic/osteogenic differentiation, and binding p53 to promote CBP/p300-mediated acetylation and p53-dependent cell-cycle arrest and apoptosis [#2, #10, #6]. It additionally competes with NRF2 for KEAP1 binding to stabilize NRF2 and protect the antioxidant response, and binds RhoGDI\\u03b1 to restrain CDC42 activation [#5, #13]. Isoform-specific functions include WTX-S driving NONO sequestration and paraspeckle disassembly via liquid-liquid phase separation, and WTX-L promoting NF-\\u03baB/LCN2-dependent ferroptosis through competitive binding to \\u03b2-arrestin2 [#15, #14].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established AMER1/WTX as a bona fide component of the \\u03b2-catenin destruction complex, defining its core role as a Wnt antagonist that promotes \\u03b2-catenin turnover.\",\n      \"evidence\": \"Tandem-affinity purification/mass spectrometry and ubiquitination/degradation assays in cells, Xenopus, and zebrafish\",\n      \"pmids\": [\"17510365\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve where in the cell the complex assembles\", \"Mechanism of how WTX promotes ubiquitination not detailed\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defined WTX as a membrane-anchored protein that recruits APC to the plasma membrane, placing destruction-complex assembly at the membrane.\",\n      \"evidence\": \"Co-IP, domain mapping of two PtdIns(4,5)P2-binding domains, siRNA, overexpression, and live imaging in multiple cell lines\",\n      \"pmids\": [\"17925383\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of APC-ARM binding not yet determined\", \"Functional consequence of membrane vs microtubule APC pools unresolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Linked WTX expression to growth suppression via p21-mediated G1/G0 arrest and mapped the responsible region, framing tumor-associated truncations as functional losses.\",\n      \"evidence\": \"Inducible expression, flow-cytometry cell-cycle analysis, truncation mutant mapping, and expression profiling\",\n      \"pmids\": [\"20956941\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular link between WTX and p21 induction not established\", \"Single inducible cell system\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed WTX directly binds \\u03b2-catenin armadillo repeats through REA motifs and stabilizes Axin at the membrane, defining the molecular interface for destruction-complex assembly.\",\n      \"evidence\": \"Co-IP, domain deletion/point mutagenesis, reporter assays, and artificial membrane-targeting rescue\",\n      \"pmids\": [\"21498506\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of the membrane destruction complex unresolved\", \"How membrane assembly couples to ubiquitination not detailed\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealed the paradoxical positive role of WTX in Wnt-induced LRP6 phosphorylation, recruiting CK1\\u03b3/GSK3\\u03b2/Axin to the membrane and producing the receptor-activating signal.\",\n      \"evidence\": \"siRNA, LRP6-Amer1 fusion construct, phosphorylation assays, and PtdIns(4,5)P2 application\",\n      \"pmids\": [\"21304492\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the same protein both activates LRP6 and degrades \\u03b2-catenin is not mechanistically reconciled\", \"Temporal switch between roles unknown\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstrated in vivo that Wtx is essential for development and that its loss drives \\u03b2-catenin\\u2013dependent mesenchymal lineage defects.\",\n      \"evidence\": \"Conditional/stage-specific mouse knockout and primary mesenchymal progenitor differentiation assays\",\n      \"pmids\": [\"21571217\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"\\u03b2-catenin-independent contributions to differentiation delay not molecularly defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified a nuclear function for WTX as a WT1 transcriptional cofactor and noted association with the paraspeckle marker NONO.\",\n      \"evidence\": \"Nuclear fractionation, Co-IP, co-localization, and WT1 target reporter assays\",\n      \"pmids\": [\"19416806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional significance of NONO co-localization not yet established\", \"Regulation of nuclear vs membrane partitioning incompletely defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed WTX inhibits rather than promotes ubiquitination in the KEAP1\\u2013NRF2 axis, competing with NRF2 to stabilize the antioxidant response and revealing context-dependent E3-adaptor effects.\",\n      \"evidence\": \"Co-IP, competition binding, ubiquitination assay, and NRF2 stability readout on WTX knockdown\",\n      \"pmids\": [\"22215675\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Why WTX promotes \\u03b2-catenin but blocks NRF2 ubiquitination not mechanistically explained\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Established WTX as a p53 cofactor that enhances CBP/p300-mediated p53 acetylation and p53-dependent arrest/apoptosis, broadening its tumor-suppressor functions.\",\n      \"evidence\": \"Domain-mapping Co-IP, p53 acetylation assays, knockdown, and p53 reconstitution with cell-cycle/apoptosis readouts\",\n      \"pmids\": [\"22285752\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How WTX stabilizes CBP/p300 not defined\", \"Crosstalk with WTX Wnt functions unexplored\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified \\u03b2-arrestin as a regulator of WTX membrane dynamics and PtdIns(4,5)P2 production during Wnt-induced LRP6 phosphorylation.\",\n      \"evidence\": \"Co-IP, FRAP, LRP6 phosphorylation assays in \\u03b2-arrestin knockout cells\",\n      \"pmids\": [\"24265322\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect coupling to PI kinases not fully resolved\", \"Single-lab characterization\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Determined the structural basis of WTX\\u2013APC recognition, showing multiple Amer1 fragments bind APC-ARM using a shared mode shared with other partners.\",\n      \"evidence\": \"X-ray crystallography, ITC, GST pull-down, yeast two-hybrid, and cell-based composite mutagenesis\",\n      \"pmids\": [\"27462415\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of WTX with \\u03b2-catenin or Axin not solved\", \"Full-length WTX architecture unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined TRIM28 as the major nuclear partner of WTX, establishing a \\u03b2-catenin-independent role in epigenetic silencing of retroelements and control of mesenchymal differentiation.\",\n      \"evidence\": \"Domain-mapping Co-IP, ChIP, single-molecule RNA-seq, and differentiation assays\",\n      \"pmids\": [\"25882849\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether WTX directly contacts chromatin unresolved\", \"How WTX is targeted to specific loci unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Explained the partial function of C-terminal WTX truncations by showing Axin can indirectly bridge \\u03b2-catenin recruitment when direct binding is lost.\",\n      \"evidence\": \"In vitro Co-IP with truncation mutants and a knock-in mouse allele\",\n      \"pmids\": [\"29329488\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Quantitative contribution of indirect bridging in vivo not measured\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended WTX's negative Wnt role to vertebrate development and regeneration and showed Amer family members do not functionally compensate.\",\n      \"evidence\": \"Zebrafish morpholino knockdown, TALEN knockout, Wnt target analysis, and fin regeneration assay\",\n      \"pmids\": [\"31290212\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Normal development of mutants despite Wnt dysregulation not reconciled with mouse lethality\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Uncovered a Wnt-independent cytoskeletal/invasion function in which WTX supports RhoGDI\\u03b1 suppression of CDC42, with loss driving metastasis.\",\n      \"evidence\": \"Co-IP, CDC42 activity assays, knockdown/overexpression, and in vivo metastasis model\",\n      \"pmids\": [\"30631060\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect WTX\\u2013RhoGDI\\u03b1 binding not structurally defined\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified KIF23 as a competitive disruptor of the Amer1\\u2013APC interaction that relocalizes Amer1 to the nucleus and activates Wnt signaling in cancer.\",\n      \"evidence\": \"Co-IP, competitive binding, subcellular relocalization, and Wnt reporter assays\",\n      \"pmids\": [\"32365332\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether nuclear-relocalized Amer1 gains nuclear function not tested\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed AMER1 downstream of IRF-2 transcriptional control, identifying an upstream regulator of its expression that constrains Wnt-driven proliferation.\",\n      \"evidence\": \"ChIP-seq, western blot, proliferation assays, and xenograft model\",\n      \"pmids\": [\"35115027\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Other transcriptional regulators of AMER1 not surveyed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined an isoform-specific WTX-L function promoting ferroptosis via competitive \\u03b2-arrestin2 binding and NF-\\u03baB/LCN2 activation.\",\n      \"evidence\": \"Competitive Co-IP, NF-\\u03baB reporter, ferroptosis inducers, and xenograft\",\n      \"pmids\": [\"40109379\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Generality beyond gastric cancer unknown\", \"How isoform choice is regulated unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Defined an isoform-specific WTX-S function that sequesters NONO into phase-separated nuclear bodies to disassemble paraspeckles and sensitize cells to chemotherapy.\",\n      \"evidence\": \"Domain-mapping Co-IP, LLPS assay, live-cell imaging, and chemosensitivity assays\",\n      \"pmids\": [\"41115290\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological triggers of WTX-S nuclear body formation unknown\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How WTX integrates its opposing roles \\u2014 destruction-complex scaffold versus LRP6-activating cofactor, and pro- versus anti-ubiquitination E3-adaptor functions \\u2014 within a single regulatory logic remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model reconciles WTX's positive and negative Wnt roles\", \"Determinants of isoform- and partner-specific function not defined\", \"Full-length structure and complex stoichiometry unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 3, 4, 8]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [5, 6, 13]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [2, 10, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 3, 4]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 10, 6]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 4, 9]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [2, 10, 6]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [9, 18]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [14, 6]}\n    ],\n    \"complexes\": [\"\\u03b2-catenin destruction complex\"],\n    \"partners\": [\"APC\", \"CTNNB1\", \"AXIN1\", \"KEAP1\", \"TP53\", \"TRIM28\", \"NONO\", \"ARRB2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}