{"gene":"CTNNAL1","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":2002,"finding":"Alpha-catulin (CTNNAL1) directly interacts with the C-terminal region of Lbc, a Rho-specific guanine nucleotide exchange factor; the required interaction regions were mapped, complex formation was detected in mammalian cells by co-fractionation and co-localization, and alpha-catulin co-expression enhances Lbc-induced GTP-Rho formation and serum response factor (SRF) activation, functioning as a scaffold for Lbc.","method":"Yeast two-hybrid screen, subcellular co-fractionation, intracellular co-localization, co-immunoprecipitation in mammalian cells, transcriptional reporter assay (SRF-luciferase), in vivo GTP-Rho pulldown assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction mapping, co-fractionation, mammalian complex formation, and two functional readouts (GTP-Rho and SRF activation) all in one study with multiple orthogonal methods","pmids":["12270917"],"is_preprint":false},{"year":2008,"finding":"CTNNAL1 is required for wound repair and proliferation of human bronchial epithelial cells (HBECs); antisense oligonucleotide (ASO) knockdown of CTNNAL1 decelerates repair velocity and proliferation, blocks fibronectin-promoted wound repair, and inhibits fibronectin-induced FAK phosphorylation, placing CTNNAL1 in the fibronectin–FAK signaling axis.","method":"Antisense oligonucleotide knockdown, scratch-wound assay, cell proliferation assay, Western blot for FAK phosphorylation","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined cellular phenotype and a downstream signaling readout (p-FAK), single lab with two orthogonal methods","pmids":["17647259"],"is_preprint":false},{"year":2012,"finding":"Transcription factors AP-2α and LEF-1 bind to the CTNNAL1 promoter and drive its transcription in human bronchial epithelial cells; binding was confirmed by EMSA, antibody supershift, and ChIP, and site-directed mutagenesis of the AP-2α and LEF-1 sites in the CTNNAL1 promoter reduced promoter activity.","method":"EMSA, antibody supershift assay, chromatin immunoprecipitation (ChIP), site-directed mutagenesis of promoter binding sites, luciferase reporter assay, antisense oligonucleotide knockdown of AP-2α and LEF-1","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple orthogonal methods (EMSA, ChIP, mutagenesis, reporter) in a single rigorous study establishing the transcriptional regulatory mechanism","pmids":["22359570"],"is_preprint":false},{"year":2018,"finding":"CTNNAL1 inhibits ozone-induced epithelial-mesenchymal transition (EMT) in airway epithelial cells; overexpression reverses EMT features, represses Twist1 mRNA expression, and reduces TGF-β1 secretion, while silencing of CTNNAL1 exacerbates ozone-induced EMT and enhances functional changes in co-cultured lung fibroblasts.","method":"Stable transfection (overexpression and siRNA silencing), ozone exposure model, EMT marker analysis, co-culture assay with lung fibroblasts, qRT-PCR for Twist1/Twist2/Snail/Slug, ELISA for TGF-β1","journal":"Experimental physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain- and loss-of-function with defined EMT phenotype and multiple molecular readouts, single lab","pmids":["29791759"],"is_preprint":false},{"year":2022,"finding":"CTNNAL1 interacts with YAP (Yes-associated protein) as confirmed by co-immunoprecipitation; CTNNAL1 silencing downregulates YAP expression and increases ROCK2, leading to mucus hypersecretion (MUC5AC) and IL-4/IL-13 elevation; YAP inhibition reduces MUC5AC, IL-4, and IL-13 and decreases ROCK2, placing CTNNAL1 upstream of a YAP–ROCK2 pathway controlling mucus secretion.","method":"AAV-delivered siRNA mouse model, co-immunoprecipitation (CTNNAL1–YAP interaction), CTNNAL1 overexpression/silencing in 16HBE14o- cells, pharmacological YAP and ROCK2 inhibition, ELISA for cytokines, PAS staining for goblet cells, MUC5AC measurement","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirms physical interaction, epistatic pathway placed with pharmacological inhibition, multiple readouts; single lab","pmids":["35092120"],"is_preprint":false},{"year":2022,"finding":"CTNNAL1 physically interacts with hsp90 (confirmed by immunoprecipitation); CTNNAL1 silencing upregulates hsp90 expression and increases NR3C1, ICAM-1, and NF-κB (p-p65) levels, reducing glucocorticoid sensitivity; hsp90 inhibitor geldanamycin rescues these effects, placing hsp90 downstream of CTNNAL1 in regulating airway inflammation and steroid responsiveness.","method":"CTNNAL1-siRNA mouse model, immunoprecipitation (CTNNAL1–hsp90), pharmacological hsp90 inhibition with geldanamycin, Western blot for NR3C1/ICAM-1/p-p65, dexamethasone treatment comparison","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirms physical interaction with hsp90, epistatic placement supported by pharmacological rescue, single lab with multiple readouts","pmids":["36535402"],"is_preprint":false},{"year":2023,"finding":"CTNNAL1 deficiency decreases CFTR expression through a ROCK1–CAL signaling axis: CTNNAL1 silencing reduces ROCK1 activity (while increasing ROCK2), ROCK1 inhibition reduces CFTR expression, RhoA activation increases CFTR, CAL (CFTR-associated ligand) expression increases upon CTNNAL1 silencing, and immunoprecipitation confirms ROCK1–CAL interaction; CAL inhibition restores CFTR expression without affecting ROCK1.","method":"CTNNAL1-siRNA mouse model, ROCK1 pharmacological inhibition and RhoA activation in HBE cells, immunoprecipitation (ROCK1–CAL interaction), Western blot for CFTR/ROCK1/ROCK2/CAL, in vitro and in vivo experiments","journal":"Acta biochimica et biophysica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP establishes ROCK1–CAL interaction, epistatic dissection with pharmacological probes, multiple orthogonal readouts; single lab","pmids":["37715489"],"is_preprint":false},{"year":2024,"finding":"CTNNAL1 maintains structural integrity of bronchial epithelial cells via the RhoA/ROCK1 pathway; CTNNAL1 knockout mice show denuded epithelial cells and airway structural damage; CTNNAL1 silencing reduces E-cadherin, integrin β1, and integrin β4 expression, weakens extracellular matrix and intercellular adhesion, and decreases RhoA/ROCK1 levels; ROCK inhibitor Y27632 abolishes ozone-induced adhesion molecule upregulation in CTNNAL1-overexpressing cells, confirming ROCK1 as the downstream effector.","method":"AAV-RNAi mouse knockout, stable siRNA-transfected HBEC cell line, HE staining, cell proliferation and adhesion assays, Western blot for E-cadherin/integrins/RhoA/ROCK1, pharmacological ROCK inhibition with Y27632, ozone stress model","journal":"Acta biochimica et biophysica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo knockout and in vitro silencing with defined structural phenotypes, epistatic rescue by ROCK inhibitor, single lab with multiple orthogonal methods","pmids":["38602002"],"is_preprint":false},{"year":2023,"finding":"CTNNAL1 regulates cancer stem cell properties, radiation resistance, migration, invasion, and CCL2 chemokine secretion in lung cancer and glioblastoma cells; loss or gain of CTNNAL1 modulates EMT markers and stem cell characteristics.","method":"CTNNAL1 overexpression and knockdown in lung cancer and glioblastoma cell lines, sphere-formation assay, irradiation resistance assay, migration/invasion assay, CCL2 ELISA","journal":"Biomedicines","confidence":"Low","confidence_rationale":"Tier 3 / Weak — defined cellular phenotypes with KD/OE but no pathway placement or mechanistic upstream/downstream identification; single lab, single study","pmids":["37239133"],"is_preprint":false}],"current_model":"CTNNAL1 (alpha-catulin) is an alpha-catenin-related scaffold protein that modulates Rho GTPase signaling by binding the Lbc Rho-GEF to enhance GTP-Rho loading and SRF activation; in airway epithelial cells it maintains structural integrity and suppresses EMT by sustaining RhoA/ROCK1-dependent adhesion molecule expression, while its loss activates a YAP–ROCK2 axis promoting mucus hypersecretion, an hsp90 pathway that reduces glucocorticoid sensitivity, and a ROCK1–CAL axis that suppresses CFTR expression; upstream, its transcription is driven by AP-2α and LEF-1 binding to its promoter."},"narrative":{"mechanistic_narrative":"CTNNAL1 (alpha-catulin) is an alpha-catenin-related scaffold protein that couples Rho GTPase signaling to the maintenance of airway epithelial integrity and the suppression of epithelial-mesenchymal transition [PMID:12270917, PMID:38602002]. It acts mechanistically as a scaffold for the Rho-specific guanine nucleotide exchange factor Lbc, binding its C-terminal region to enhance GTP-Rho loading and downstream SRF activation [PMID:12270917]. In bronchial epithelium CTNNAL1 sustains a RhoA/ROCK1-dependent program that maintains expression of E-cadherin and integrins and thereby preserves intercellular and matrix adhesion, with its knockout producing denuded epithelium and structural airway damage [PMID:38602002]; consistent with this adhesive/repair role, it is required for fibronectin-promoted wound repair and FAK phosphorylation [PMID:17647259] and represses ozone-induced EMT by lowering Twist1 and TGF-β1 [PMID:29791759]. Loss of CTNNAL1 diverts signaling toward pathological outputs: a YAP–ROCK2 axis driving MUC5AC mucus hypersecretion and IL-4/IL-13 elevation [PMID:35092120], an hsp90 axis that raises NR3C1, ICAM-1, and NF-κB and reduces glucocorticoid sensitivity [PMID:36535402], and a ROCK1–CAL axis that suppresses CFTR expression [PMID:37715489]. Its own transcription is driven by AP-2α and LEF-1 binding to its promoter [PMID:22359570].","teleology":[{"year":2002,"claim":"Established the first molecular function of CTNNAL1: it is not merely an adhesion-related catenin homolog but a scaffold that links a Rho-GEF to active Rho/SRF signaling.","evidence":"Yeast two-hybrid, interaction-region mapping, co-fractionation/co-localization, GTP-Rho pulldown, and SRF-luciferase reporter in mammalian cells","pmids":["12270917"],"confidence":"High","gaps":["Does not define which cell types depend on the Lbc scaffold function in vivo","No structural model of the CTNNAL1–Lbc complex","Link between this Rho-GEF scaffolding and the later RhoA/ROCK epithelial phenotypes not directly traced"]},{"year":2008,"claim":"Placed CTNNAL1 in epithelial repair, showing it is required for proliferation, wound closure, and fibronectin-induced FAK activation.","evidence":"Antisense knockdown with scratch-wound, proliferation assays, and p-FAK Western blot in human bronchial epithelial cells","pmids":["17647259"],"confidence":"Medium","gaps":["Does not establish whether the FAK effect is direct or via Rho signaling","Mechanism of fibronectin–CTNNAL1 coupling unresolved"]},{"year":2012,"claim":"Defined the upstream transcriptional control of CTNNAL1, identifying AP-2α and LEF-1 as direct promoter-binding activators.","evidence":"EMSA, supershift, ChIP, promoter mutagenesis, and luciferase reporter in bronchial epithelial cells","pmids":["22359570"],"confidence":"High","gaps":["Does not connect transcriptional regulation to a physiological stimulus","No signaling input shown to control AP-2α/LEF-1 occupancy of the CTNNAL1 promoter"]},{"year":2018,"claim":"Showed CTNNAL1 is a suppressor of EMT, repressing Twist1 and TGF-β1 to protect airway epithelium from ozone-induced mesenchymal transition.","evidence":"Overexpression and siRNA in airway epithelial cells under ozone exposure, EMT marker qRT-PCR, fibroblast co-culture, TGF-β1 ELISA","pmids":["29791759"],"confidence":"Medium","gaps":["Pathway linking CTNNAL1 to Twist1/TGF-β1 repression not defined","Single-lab cellular model"]},{"year":2022,"claim":"Identified a YAP–ROCK2 axis downstream of CTNNAL1 loss, explaining how its deficiency drives mucus hypersecretion and type-2 cytokine elevation.","evidence":"AAV-siRNA mouse model, CTNNAL1–YAP co-IP, gain/loss in 16HBE14o- cells, pharmacological YAP/ROCK2 inhibition, MUC5AC and cytokine readouts","pmids":["35092120"],"confidence":"Medium","gaps":["Direct vs indirect nature of CTNNAL1–YAP binding not structurally resolved","How CTNNAL1 loss shifts ROCK1→ROCK2 balance unexplained"]},{"year":2022,"claim":"Connected CTNNAL1 to glucocorticoid responsiveness through an hsp90 axis controlling NR3C1, ICAM-1, and NF-κB.","evidence":"CTNNAL1-siRNA mouse model, CTNNAL1–hsp90 immunoprecipitation, geldanamycin rescue, Western blots, dexamethasone comparison","pmids":["36535402"],"confidence":"Medium","gaps":["Whether CTNNAL1 binding regulates hsp90 chaperone activity directly is untested","Reciprocal interaction validation not shown"]},{"year":2023,"claim":"Dissected a ROCK1–CAL axis whereby CTNNAL1 deficiency suppresses CFTR, linking the protein to ion-channel regulation in airway disease.","evidence":"CTNNAL1-siRNA mouse model, ROCK1 inhibition and RhoA activation in HBE cells, ROCK1–CAL immunoprecipitation, CFTR/ROCK Western blots","pmids":["37715489"],"confidence":"Medium","gaps":["Mechanism by which CTNNAL1 sustains ROCK1 over ROCK2 not defined","Single-lab finding"]},{"year":2023,"claim":"Extended CTNNAL1 function to cancer, implicating it in stem-cell properties, radioresistance, invasion, and CCL2 secretion in lung cancer and glioblastoma.","evidence":"Overexpression/knockdown in lung cancer and glioblastoma cell lines with sphere-formation, irradiation, migration/invasion assays, CCL2 ELISA","pmids":["37239133"],"confidence":"Low","gaps":["No pathway placement or upstream/downstream mechanism identified","Single lab, single study","Relationship to the airway Rho/ROCK functions untested"]},{"year":2024,"claim":"Consolidated the structural role of CTNNAL1, showing it maintains epithelial adhesion molecule expression and integrity via RhoA/ROCK1 in vivo.","evidence":"AAV-RNAi knockout mice, stable siRNA HBEC line, HE staining, adhesion assays, ROCK inhibitor Y27632 rescue, Western blots for E-cadherin/integrins/RhoA/ROCK1","pmids":["38602002"],"confidence":"Medium","gaps":["Does not resolve how the Lbc-scaffolding activity feeds into RhoA/ROCK1 in epithelium","Direct CTNNAL1 binding partners in the adhesion complex not mapped"]},{"year":null,"claim":"How the single biochemical activity of CTNNAL1 (Lbc/Rho-GEF scaffolding) mechanistically determines the divergent ROCK1 vs ROCK2 outputs and its multiple binding partners (YAP, hsp90, ROCK1–CAL) remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model linking scaffold function to downstream effector selection","Whether YAP, hsp90, and CAL bind CTNNAL1 directly or via Rho-pathway intermediates is unclear","No reconstitution of CTNNAL1's role in ROCK1/ROCK2 balance"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,7]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[3,7]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[2]}],"complexes":[],"partners":["LBC/AKAP13","YAP1","HSP90","ROCK1","GOPC"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UBT7","full_name":"Alpha-catulin","aliases":["Alpha-catenin-related protein","ACRP","Catenin alpha-like protein 1"],"length_aa":734,"mass_kda":81.9,"function":"May modulate the Rho pathway signaling by providing a scaffold for the Lbc Rho guanine nucleotide exchange factor (ARHGEF1)","subcellular_location":"Cytoplasm, cytoskeleton; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9UBT7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CTNNAL1","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":[{"gene":"UTRN","stoichiometry":4.0},{"gene":"DHX9","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/CTNNAL1","total_profiled":1310},"omim":[{"mim_id":"608026","title":"HYPERTENSIVE NEPHROPATHY; HNP1","url":"https://www.omim.org/entry/608026"},{"mim_id":"607667","title":"CATENIN, ALPHA-3; CTNNA3","url":"https://www.omim.org/entry/607667"},{"mim_id":"604785","title":"CATENIN, ALPHA-LIKE, 1; CTNNAL1","url":"https://www.omim.org/entry/604785"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"adrenal gland","ntpm":273.5}],"url":"https://www.proteinatlas.org/search/CTNNAL1"},"hgnc":{"alias_symbol":["CLLP","alpha-CATU"],"prev_symbol":[]},"alphafold":{"accession":"Q9UBT7","domains":[{"cath_id":"1.20.120.230","chopping":"34-120_136-160","consensus_level":"high","plddt":85.6529,"start":34,"end":160},{"cath_id":"1.20.120.230","chopping":"168-276","consensus_level":"medium","plddt":96.1094,"start":168,"end":276},{"cath_id":"1.20.120.330","chopping":"291-402","consensus_level":"medium","plddt":91.7314,"start":291,"end":402},{"cath_id":"1.20.120.230","chopping":"558-718","consensus_level":"high","plddt":87.6623,"start":558,"end":718}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UBT7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UBT7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UBT7-F1-predicted_aligned_error_v6.png","plddt_mean":84.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CTNNAL1","jax_strain_url":"https://www.jax.org/strain/search?query=CTNNAL1"},"sequence":{"accession":"Q9UBT7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9UBT7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9UBT7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UBT7"}},"corpus_meta":[{"pmid":"12270917","id":"PMC_12270917","title":"Association of Lbc Rho guanine nucleotide exchange factor with alpha-catenin-related protein, alpha-catulin/CTNNAL1, supports serum response factor activation.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12270917","citation_count":53,"is_preprint":false},{"pmid":"9806841","id":"PMC_9806841","title":"Identification and chromosomal localization of CTNNAL1, a novel protein homologous to alpha-catenin.","date":"1998","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/9806841","citation_count":33,"is_preprint":false},{"pmid":"17647259","id":"PMC_17647259","title":"Wound repair and proliferation of bronchial epithelial cells regulated by CTNNAL1.","date":"2008","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17647259","citation_count":22,"is_preprint":false},{"pmid":"23820070","id":"PMC_23820070","title":"Associations of TCF12, CTNNAL1 and WNT10B gene polymorphisms with litter size in pigs.","date":"2013","source":"Animal reproduction science","url":"https://pubmed.ncbi.nlm.nih.gov/23820070","citation_count":17,"is_preprint":false},{"pmid":"35092120","id":"PMC_35092120","title":"CTNNAL1 participates in the regulation of mucus overproduction in HDM-induced asthma mouse model through the YAP-ROCK2 pathway.","date":"2022","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35092120","citation_count":14,"is_preprint":false},{"pmid":"29791759","id":"PMC_29791759","title":"CTNNAL1 inhibits ozone-induced epithelial-mesenchymal transition in human bronchial epithelial cells.","date":"2018","source":"Experimental physiology","url":"https://pubmed.ncbi.nlm.nih.gov/29791759","citation_count":13,"is_preprint":false},{"pmid":"22359570","id":"PMC_22359570","title":"Identification of transcription factors regulating CTNNAL1 expression in human bronchial epithelial cells.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22359570","citation_count":13,"is_preprint":false},{"pmid":"30178695","id":"PMC_30178695","title":"Genetic effect and combined genotype effect of ESR, FSHβ, CTNNAL1 and miR-27a loci on litter size in a Large White population.","date":"2018","source":"Animal biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/30178695","citation_count":11,"is_preprint":false},{"pmid":"32139661","id":"PMC_32139661","title":"Genetic variants in RET, ARHGEF3 and CTNNAL1, and relevant interaction networks, contribute to the risk of Hirschsprung disease.","date":"2020","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/32139661","citation_count":9,"is_preprint":false},{"pmid":"37239133","id":"PMC_37239133","title":"Regulation of Cancer Stem Cells and Epithelial-Mesenchymal Transition by CTNNAL1 in Lung Cancer and Glioblastoma.","date":"2023","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/37239133","citation_count":7,"is_preprint":false},{"pmid":"36535402","id":"PMC_36535402","title":"CTNNAL1 enhances glucocorticoid sensitivity in HDM-induced asthma mouse model through deactivating hsp90 signaling pathway.","date":"2022","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36535402","citation_count":4,"is_preprint":false},{"pmid":"38602002","id":"PMC_38602002","title":"CTNNAL1 promotes the structural integrity of bronchial epithelial cells through the RhoA/ROCK1 pathway.","date":"2024","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/38602002","citation_count":2,"is_preprint":false},{"pmid":"23000764","id":"PMC_23000764","title":"[Relevance of the expression of CTNNAL1 and the resistance of respiratory tract in rat with airway hyperresponsiveness].","date":"2012","source":"Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/23000764","citation_count":2,"is_preprint":false},{"pmid":"37715489","id":"PMC_37715489","title":"CTNNAL1 deficiency suppresses CFTR expression in HDM-induced asthma mouse model through ROCK1-CAL signaling pathway.","date":"2023","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/37715489","citation_count":1,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.06.12.25329506","title":"Genome-wide association study of pediatric bacteremia and sepsis","date":"2025-06-12","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.12.25329506","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10857,"output_tokens":2848,"usd":0.037645,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10254,"output_tokens":3450,"usd":0.06876,"stage2_stop_reason":"end_turn"},"total_usd":0.106405,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"Alpha-catulin (CTNNAL1) directly interacts with the C-terminal region of Lbc, a Rho-specific guanine nucleotide exchange factor; the required interaction regions were mapped, complex formation was detected in mammalian cells by co-fractionation and co-localization, and alpha-catulin co-expression enhances Lbc-induced GTP-Rho formation and serum response factor (SRF) activation, functioning as a scaffold for Lbc.\",\n      \"method\": \"Yeast two-hybrid screen, subcellular co-fractionation, intracellular co-localization, co-immunoprecipitation in mammalian cells, transcriptional reporter assay (SRF-luciferase), in vivo GTP-Rho pulldown assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction mapping, co-fractionation, mammalian complex formation, and two functional readouts (GTP-Rho and SRF activation) all in one study with multiple orthogonal methods\",\n      \"pmids\": [\"12270917\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"CTNNAL1 is required for wound repair and proliferation of human bronchial epithelial cells (HBECs); antisense oligonucleotide (ASO) knockdown of CTNNAL1 decelerates repair velocity and proliferation, blocks fibronectin-promoted wound repair, and inhibits fibronectin-induced FAK phosphorylation, placing CTNNAL1 in the fibronectin–FAK signaling axis.\",\n      \"method\": \"Antisense oligonucleotide knockdown, scratch-wound assay, cell proliferation assay, Western blot for FAK phosphorylation\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined cellular phenotype and a downstream signaling readout (p-FAK), single lab with two orthogonal methods\",\n      \"pmids\": [\"17647259\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Transcription factors AP-2α and LEF-1 bind to the CTNNAL1 promoter and drive its transcription in human bronchial epithelial cells; binding was confirmed by EMSA, antibody supershift, and ChIP, and site-directed mutagenesis of the AP-2α and LEF-1 sites in the CTNNAL1 promoter reduced promoter activity.\",\n      \"method\": \"EMSA, antibody supershift assay, chromatin immunoprecipitation (ChIP), site-directed mutagenesis of promoter binding sites, luciferase reporter assay, antisense oligonucleotide knockdown of AP-2α and LEF-1\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple orthogonal methods (EMSA, ChIP, mutagenesis, reporter) in a single rigorous study establishing the transcriptional regulatory mechanism\",\n      \"pmids\": [\"22359570\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CTNNAL1 inhibits ozone-induced epithelial-mesenchymal transition (EMT) in airway epithelial cells; overexpression reverses EMT features, represses Twist1 mRNA expression, and reduces TGF-β1 secretion, while silencing of CTNNAL1 exacerbates ozone-induced EMT and enhances functional changes in co-cultured lung fibroblasts.\",\n      \"method\": \"Stable transfection (overexpression and siRNA silencing), ozone exposure model, EMT marker analysis, co-culture assay with lung fibroblasts, qRT-PCR for Twist1/Twist2/Snail/Slug, ELISA for TGF-β1\",\n      \"journal\": \"Experimental physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain- and loss-of-function with defined EMT phenotype and multiple molecular readouts, single lab\",\n      \"pmids\": [\"29791759\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CTNNAL1 interacts with YAP (Yes-associated protein) as confirmed by co-immunoprecipitation; CTNNAL1 silencing downregulates YAP expression and increases ROCK2, leading to mucus hypersecretion (MUC5AC) and IL-4/IL-13 elevation; YAP inhibition reduces MUC5AC, IL-4, and IL-13 and decreases ROCK2, placing CTNNAL1 upstream of a YAP–ROCK2 pathway controlling mucus secretion.\",\n      \"method\": \"AAV-delivered siRNA mouse model, co-immunoprecipitation (CTNNAL1–YAP interaction), CTNNAL1 overexpression/silencing in 16HBE14o- cells, pharmacological YAP and ROCK2 inhibition, ELISA for cytokines, PAS staining for goblet cells, MUC5AC measurement\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirms physical interaction, epistatic pathway placed with pharmacological inhibition, multiple readouts; single lab\",\n      \"pmids\": [\"35092120\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CTNNAL1 physically interacts with hsp90 (confirmed by immunoprecipitation); CTNNAL1 silencing upregulates hsp90 expression and increases NR3C1, ICAM-1, and NF-κB (p-p65) levels, reducing glucocorticoid sensitivity; hsp90 inhibitor geldanamycin rescues these effects, placing hsp90 downstream of CTNNAL1 in regulating airway inflammation and steroid responsiveness.\",\n      \"method\": \"CTNNAL1-siRNA mouse model, immunoprecipitation (CTNNAL1–hsp90), pharmacological hsp90 inhibition with geldanamycin, Western blot for NR3C1/ICAM-1/p-p65, dexamethasone treatment comparison\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirms physical interaction with hsp90, epistatic placement supported by pharmacological rescue, single lab with multiple readouts\",\n      \"pmids\": [\"36535402\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CTNNAL1 deficiency decreases CFTR expression through a ROCK1–CAL signaling axis: CTNNAL1 silencing reduces ROCK1 activity (while increasing ROCK2), ROCK1 inhibition reduces CFTR expression, RhoA activation increases CFTR, CAL (CFTR-associated ligand) expression increases upon CTNNAL1 silencing, and immunoprecipitation confirms ROCK1–CAL interaction; CAL inhibition restores CFTR expression without affecting ROCK1.\",\n      \"method\": \"CTNNAL1-siRNA mouse model, ROCK1 pharmacological inhibition and RhoA activation in HBE cells, immunoprecipitation (ROCK1–CAL interaction), Western blot for CFTR/ROCK1/ROCK2/CAL, in vitro and in vivo experiments\",\n      \"journal\": \"Acta biochimica et biophysica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP establishes ROCK1–CAL interaction, epistatic dissection with pharmacological probes, multiple orthogonal readouts; single lab\",\n      \"pmids\": [\"37715489\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CTNNAL1 maintains structural integrity of bronchial epithelial cells via the RhoA/ROCK1 pathway; CTNNAL1 knockout mice show denuded epithelial cells and airway structural damage; CTNNAL1 silencing reduces E-cadherin, integrin β1, and integrin β4 expression, weakens extracellular matrix and intercellular adhesion, and decreases RhoA/ROCK1 levels; ROCK inhibitor Y27632 abolishes ozone-induced adhesion molecule upregulation in CTNNAL1-overexpressing cells, confirming ROCK1 as the downstream effector.\",\n      \"method\": \"AAV-RNAi mouse knockout, stable siRNA-transfected HBEC cell line, HE staining, cell proliferation and adhesion assays, Western blot for E-cadherin/integrins/RhoA/ROCK1, pharmacological ROCK inhibition with Y27632, ozone stress model\",\n      \"journal\": \"Acta biochimica et biophysica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockout and in vitro silencing with defined structural phenotypes, epistatic rescue by ROCK inhibitor, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"38602002\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CTNNAL1 regulates cancer stem cell properties, radiation resistance, migration, invasion, and CCL2 chemokine secretion in lung cancer and glioblastoma cells; loss or gain of CTNNAL1 modulates EMT markers and stem cell characteristics.\",\n      \"method\": \"CTNNAL1 overexpression and knockdown in lung cancer and glioblastoma cell lines, sphere-formation assay, irradiation resistance assay, migration/invasion assay, CCL2 ELISA\",\n      \"journal\": \"Biomedicines\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — defined cellular phenotypes with KD/OE but no pathway placement or mechanistic upstream/downstream identification; single lab, single study\",\n      \"pmids\": [\"37239133\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CTNNAL1 (alpha-catulin) is an alpha-catenin-related scaffold protein that modulates Rho GTPase signaling by binding the Lbc Rho-GEF to enhance GTP-Rho loading and SRF activation; in airway epithelial cells it maintains structural integrity and suppresses EMT by sustaining RhoA/ROCK1-dependent adhesion molecule expression, while its loss activates a YAP–ROCK2 axis promoting mucus hypersecretion, an hsp90 pathway that reduces glucocorticoid sensitivity, and a ROCK1–CAL axis that suppresses CFTR expression; upstream, its transcription is driven by AP-2α and LEF-1 binding to its promoter.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CTNNAL1 (alpha-catulin) is an alpha-catenin-related scaffold protein that couples Rho GTPase signaling to the maintenance of airway epithelial integrity and the suppression of epithelial-mesenchymal transition [#0, #7]. It acts mechanistically as a scaffold for the Rho-specific guanine nucleotide exchange factor Lbc, binding its C-terminal region to enhance GTP-Rho loading and downstream SRF activation [#0]. In bronchial epithelium CTNNAL1 sustains a RhoA/ROCK1-dependent program that maintains expression of E-cadherin and integrins and thereby preserves intercellular and matrix adhesion, with its knockout producing denuded epithelium and structural airway damage [#7]; consistent with this adhesive/repair role, it is required for fibronectin-promoted wound repair and FAK phosphorylation [#1] and represses ozone-induced EMT by lowering Twist1 and TGF-\\u03b21 [#3]. Loss of CTNNAL1 diverts signaling toward pathological outputs: a YAP\\u2013ROCK2 axis driving MUC5AC mucus hypersecretion and IL-4/IL-13 elevation [#4], an hsp90 axis that raises NR3C1, ICAM-1, and NF-\\u03baB and reduces glucocorticoid sensitivity [#5], and a ROCK1\\u2013CAL axis that suppresses CFTR expression [#6]. Its own transcription is driven by AP-2\\u03b1 and LEF-1 binding to its promoter [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Established the first molecular function of CTNNAL1: it is not merely an adhesion-related catenin homolog but a scaffold that links a Rho-GEF to active Rho/SRF signaling.\",\n      \"evidence\": \"Yeast two-hybrid, interaction-region mapping, co-fractionation/co-localization, GTP-Rho pulldown, and SRF-luciferase reporter in mammalian cells\",\n      \"pmids\": [\"12270917\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not define which cell types depend on the Lbc scaffold function in vivo\", \"No structural model of the CTNNAL1\\u2013Lbc complex\", \"Link between this Rho-GEF scaffolding and the later RhoA/ROCK epithelial phenotypes not directly traced\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Placed CTNNAL1 in epithelial repair, showing it is required for proliferation, wound closure, and fibronectin-induced FAK activation.\",\n      \"evidence\": \"Antisense knockdown with scratch-wound, proliferation assays, and p-FAK Western blot in human bronchial epithelial cells\",\n      \"pmids\": [\"17647259\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not establish whether the FAK effect is direct or via Rho signaling\", \"Mechanism of fibronectin\\u2013CTNNAL1 coupling unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined the upstream transcriptional control of CTNNAL1, identifying AP-2\\u03b1 and LEF-1 as direct promoter-binding activators.\",\n      \"evidence\": \"EMSA, supershift, ChIP, promoter mutagenesis, and luciferase reporter in bronchial epithelial cells\",\n      \"pmids\": [\"22359570\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not connect transcriptional regulation to a physiological stimulus\", \"No signaling input shown to control AP-2\\u03b1/LEF-1 occupancy of the CTNNAL1 promoter\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showed CTNNAL1 is a suppressor of EMT, repressing Twist1 and TGF-\\u03b21 to protect airway epithelium from ozone-induced mesenchymal transition.\",\n      \"evidence\": \"Overexpression and siRNA in airway epithelial cells under ozone exposure, EMT marker qRT-PCR, fibroblast co-culture, TGF-\\u03b21 ELISA\",\n      \"pmids\": [\"29791759\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Pathway linking CTNNAL1 to Twist1/TGF-\\u03b21 repression not defined\", \"Single-lab cellular model\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified a YAP\\u2013ROCK2 axis downstream of CTNNAL1 loss, explaining how its deficiency drives mucus hypersecretion and type-2 cytokine elevation.\",\n      \"evidence\": \"AAV-siRNA mouse model, CTNNAL1\\u2013YAP co-IP, gain/loss in 16HBE14o- cells, pharmacological YAP/ROCK2 inhibition, MUC5AC and cytokine readouts\",\n      \"pmids\": [\"35092120\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect nature of CTNNAL1\\u2013YAP binding not structurally resolved\", \"How CTNNAL1 loss shifts ROCK1\\u2192ROCK2 balance unexplained\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Connected CTNNAL1 to glucocorticoid responsiveness through an hsp90 axis controlling NR3C1, ICAM-1, and NF-\\u03baB.\",\n      \"evidence\": \"CTNNAL1-siRNA mouse model, CTNNAL1\\u2013hsp90 immunoprecipitation, geldanamycin rescue, Western blots, dexamethasone comparison\",\n      \"pmids\": [\"36535402\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether CTNNAL1 binding regulates hsp90 chaperone activity directly is untested\", \"Reciprocal interaction validation not shown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Dissected a ROCK1\\u2013CAL axis whereby CTNNAL1 deficiency suppresses CFTR, linking the protein to ion-channel regulation in airway disease.\",\n      \"evidence\": \"CTNNAL1-siRNA mouse model, ROCK1 inhibition and RhoA activation in HBE cells, ROCK1\\u2013CAL immunoprecipitation, CFTR/ROCK Western blots\",\n      \"pmids\": [\"37715489\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which CTNNAL1 sustains ROCK1 over ROCK2 not defined\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended CTNNAL1 function to cancer, implicating it in stem-cell properties, radioresistance, invasion, and CCL2 secretion in lung cancer and glioblastoma.\",\n      \"evidence\": \"Overexpression/knockdown in lung cancer and glioblastoma cell lines with sphere-formation, irradiation, migration/invasion assays, CCL2 ELISA\",\n      \"pmids\": [\"37239133\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No pathway placement or upstream/downstream mechanism identified\", \"Single lab, single study\", \"Relationship to the airway Rho/ROCK functions untested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Consolidated the structural role of CTNNAL1, showing it maintains epithelial adhesion molecule expression and integrity via RhoA/ROCK1 in vivo.\",\n      \"evidence\": \"AAV-RNAi knockout mice, stable siRNA HBEC line, HE staining, adhesion assays, ROCK inhibitor Y27632 rescue, Western blots for E-cadherin/integrins/RhoA/ROCK1\",\n      \"pmids\": [\"38602002\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not resolve how the Lbc-scaffolding activity feeds into RhoA/ROCK1 in epithelium\", \"Direct CTNNAL1 binding partners in the adhesion complex not mapped\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the single biochemical activity of CTNNAL1 (Lbc/Rho-GEF scaffolding) mechanistically determines the divergent ROCK1 vs ROCK2 outputs and its multiple binding partners (YAP, hsp90, ROCK1\\u2013CAL) remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model linking scaffold function to downstream effector selection\", \"Whether YAP, hsp90, and CAL bind CTNNAL1 directly or via Rho-pathway intermediates is unclear\", \"No reconstitution of CTNNAL1's role in ROCK1/ROCK2 balance\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 7]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [3, 7]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"Lbc/AKAP13\", \"YAP1\", \"HSP90\", \"ROCK1\", \"GOPC\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}