{"gene":"NOP14","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2001,"finding":"NOP14 is a nucleolar protein required for 40S ribosome biogenesis; yeast depleted of Nop14 show selectively reduced levels of 20S pre-rRNA and mature 18S rRNA and diminished cellular levels of the 40S ribosomal subunit.","method":"Yeast depletion experiments, Northern blot analysis of pre-rRNA processing intermediates","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean loss-of-function with specific pre-rRNA processing phenotype, replicated in mammalian ortholog context, multiple orthogonal methods","pmids":["11694595"],"is_preprint":false},{"year":2001,"finding":"EMG1 nuclear localization depends on its physical interaction with NOP14; the two proteins interact directly, and this interaction is required for 18S rRNA maturation and 40S ribosome production.","method":"Co-immunoprecipitation, subcellular fractionation/localization, genetic depletion in yeast","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction established with localization consequence, multiple orthogonal methods (Co-IP + localization + rRNA processing assay)","pmids":["11694595"],"is_preprint":false},{"year":2017,"finding":"NOP14 increases the stability of mutant p53 (mutp53) mRNA, and the NOP14/mutp53 axis suppresses p21 expression at both transcriptional and post-transcriptional levels via induction of miR-17-5p in pancreatic cancer cells.","method":"NOP14 knockdown/overexpression, mRNA stability assay, Western blot, in vivo tumor mouse models (subcutaneous, orthotopic, intravenous injection)","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple cellular assays and in vivo models in a single lab; mRNA stability mechanistic link established but not independently replicated","pmids":["28280038"],"is_preprint":false},{"year":2015,"finding":"NOP14 enhances ERα expression and inhibits the Wnt/β-catenin pathway by up-regulating NRIP1 expression in breast cancer cells.","method":"NOP14 overexpression/knockdown, Western blot, in vivo and in vitro tumor assays","journal":"Oncotarget","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement via OE/KD with protein level readouts but no direct mechanistic assay linking NOP14 to NRIP1 regulation","pmids":["26213846"],"is_preprint":false},{"year":2021,"finding":"NOP14 promotes colorectal cancer cell growth, migration, and invasion by activating NRIP1 expression and promoting phosphorylation-inactivation of GSK-3β, leading to upregulation of β-catenin (NRIP1/GSK-3β/β-catenin signaling pathway).","method":"GFP-NOP14 overexpression, siRNA knockdown, Western blot, Transwell migration assay, CCK-8, flow cytometry","journal":"European journal of histochemistry","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single set of methods, pathway placement by OE/KD without direct biochemical reconstitution","pmids":["34218653"],"is_preprint":false},{"year":2021,"finding":"Inhibition of NOP14 (via vioprolide A treatment or NOP14 knockdown) downregulates importin-dependent NF-κB p65 nuclear translocation, reducing NF-κB promoter activity and inflammatory gene expression in endothelial cells; NOP14 knockdown confirmed a causal link between NOP14 and the anti-inflammatory action of vioprolide A.","method":"NOP14 knockdown (siRNA), vioprolide A pharmacological inhibition, NF-κB reporter assay, nuclear translocation imaging, in vivo leukocyte trafficking (cremaster muscle) and choroidal neovascularization mouse models","journal":"Biomedicine & pharmacotherapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological and genetic approaches converge on same mechanism, in vivo and in vitro validation, single lab","pmids":["34607110"],"is_preprint":false},{"year":2024,"finding":"NOP14, as a 40S ribosome biogenesis factor and target of the mTORC1-S6K axis, is required for mTORC2 activation and Akt stabilization; the mTORC2 complex is recruited to the rough endoplasmic reticulum through association with ER-bound ribosomes, and NOP14 knockdown leads to mTORC2 inactivation and Akt destabilization.","method":"NOP14 knockdown/overexpression, subcellular fractionation, co-immunoprecipitation, Western blot (phospho-Akt), cancer cell lines","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — fractionation and Co-IP establish spatial mechanism, multiple orthogonal methods in single lab; functional consequence (Akt destabilization) confirmed with loss-of-function","pmids":["38272224"],"is_preprint":false},{"year":2020,"finding":"EMG1 physically interacts with NOP14 (confirmed by GST pulldown and co-immunoprecipitation), and co-overexpression of EMG1 and NOP14 cooperatively decreases levels of WNT3a, β-catenin, phospho-GSK-3β, and c-Myc in melanoma cells.","method":"GST pulldown, co-immunoprecipitation, overexpression in melanoma cell lines, Western blot","journal":"Translational cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal binding confirmed by two orthogonal pulldown methods; functional consequence in cells; single lab","pmids":["35117729"],"is_preprint":false},{"year":2022,"finding":"NOP14 overexpression inactivates Wnt/β-catenin signaling in melanoma stem-like cells (CD133+), reducing stemness markers (Nanog, SOX2, OCT4), colony-forming ability, and the proportion of CD133+ cells; the Wnt activator BML-284 rescued these effects, placing NOP14 upstream of Wnt/β-catenin in this context.","method":"NOP14 overexpression (lentiviral), Wnt activator rescue (BML-284), flow cytometry, colony formation assay, Western blot in A375 and A875 melanoma cells","journal":"Bioengineered","confidence":"Low","confidence_rationale":"Tier 3 / Weak — epistasis via pharmacological rescue, single lab, no direct biochemical mechanism linking NOP14 to Wnt components","pmids":["35282769"],"is_preprint":false},{"year":2026,"finding":"NOP14 activates Wnt/β-catenin signaling in retinal endothelial cells (increased p-GSK-3β, β-catenin, Cyclin D1), promoting ribosome biogenesis and endothelial proliferation; NOP14 knockdown suppresses these effects and reduces angiogenesis-related proteins (CD31, VEGFA, PDGF, ANG2) in a PDR mouse model.","method":"NOP14 knockdown/overexpression, Wnt/β-catenin reporter assay, Western blot, in vivo PDR mouse model, in vitro HG-treated HRECs","journal":"Experimental cell research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, mechanistic link to Wnt pathway inferred from protein level changes without direct biochemical reconstitution","pmids":["41638389"],"is_preprint":false}],"current_model":"NOP14 is an evolutionarily conserved nucleolar protein that functions as a 40S ribosome biogenesis factor by interacting with EMG1 (required for EMG1 nuclear localization) and is required for 18S rRNA maturation; beyond ribosome assembly, NOP14 acts downstream of mTORC1-S6K signaling to support mTORC2 activation and Akt stability by facilitating recruitment of mTORC2 to ER-bound ribosomes, modulates NF-κB p65 nuclear translocation through importin-dependent mechanisms, stabilizes mutant p53 mRNA to drive oncogenic signaling, and regulates the Wnt/β-catenin pathway (through NRIP1/GSK-3β) in multiple cancer contexts."},"narrative":{"mechanistic_narrative":"NOP14 is an evolutionarily conserved nucleolar ribosome biogenesis factor required for production of the small (40S) ribosomal subunit, with depletion selectively impairing processing of 20S pre-rRNA to mature 18S rRNA [PMID:11694595]. It executes this function through a direct physical interaction with EMG1, which it retains in the nucleus; loss of this NOP14–EMG1 association blocks 18S rRNA maturation and 40S subunit assembly [PMID:11694595, PMID:35117729]. Beyond its core role in ribosome assembly, NOP14 couples ribosome biogenesis to growth signaling: acting downstream of the mTORC1–S6K axis, it is required for recruitment of mTORC2 to ER-bound ribosomes, supporting mTORC2 activation and Akt stabilization [PMID:38272224]. NOP14 also governs importin-dependent nuclear translocation of NF-κB p65, controlling inflammatory gene expression in endothelial cells [PMID:34607110], and stabilizes mutant p53 mRNA to suppress p21 via miR-17-5p induction in pancreatic cancer [PMID:28280038]. Across multiple cancer and vascular contexts NOP14 modulates Wnt/β-catenin signaling through NRIP1 and GSK-3β, though the direction of this effect is context-dependent [PMID:35117729].","teleology":[{"year":2001,"claim":"Established NOP14's core cellular function by showing it is a nucleolar factor essential for the small ribosomal subunit, answering what process the protein serves.","evidence":"Yeast depletion with Northern analysis of pre-rRNA processing intermediates and 40S subunit levels","pmids":["11694595"],"confidence":"High","gaps":["Does not define the molecular activity of NOP14 within the processing machinery","Structural basis for pre-rRNA processing role not resolved"]},{"year":2001,"claim":"Identified EMG1 as a direct NOP14 partner whose nuclear localization depends on NOP14, linking the interaction mechanistically to 18S maturation.","evidence":"Co-immunoprecipitation, subcellular fractionation, and genetic depletion in yeast","pmids":["11694595"],"confidence":"High","gaps":["Interaction interface not mapped","Whether other 40S assembly factors are recruited by NOP14 unknown"]},{"year":2015,"claim":"First placed NOP14 in a cancer signaling context, linking it to ERα and Wnt/β-catenin via NRIP1 upregulation.","evidence":"Overexpression/knockdown with Western blot and tumor assays in breast cancer cells","pmids":["26213846"],"confidence":"Low","gaps":["No direct mechanistic assay connecting NOP14 to NRIP1 regulation","Single lab, protein-level readouts only"]},{"year":2017,"claim":"Revealed a non-ribosomal RNA-stability function, showing NOP14 stabilizes mutant p53 mRNA to drive a p21-suppressing miR-17-5p axis.","evidence":"Knockdown/overexpression, mRNA stability assays, and in vivo pancreatic cancer models","pmids":["28280038"],"confidence":"Medium","gaps":["Direct RNA-binding by NOP14 not demonstrated","Not independently replicated","Mechanism of mRNA stabilization unknown"]},{"year":2020,"claim":"Confirmed the NOP14–EMG1 interaction biochemically in human cells and tied co-expression to suppression of Wnt/β-catenin signaling components.","evidence":"GST pulldown, co-immunoprecipitation, and overexpression in melanoma cells","pmids":["35117729"],"confidence":"Medium","gaps":["Functional link to Wnt is correlative protein-level","Whether ribosome biogenesis role mediates the Wnt effect untested"]},{"year":2021,"claim":"Defined NOP14 as a controller of importin-dependent NF-κB p65 nuclear translocation and the target of the anti-inflammatory compound vioprolide A.","evidence":"siRNA knockdown, pharmacological inhibition, NF-κB reporter and translocation imaging, and in vivo models","pmids":["34607110"],"confidence":"Medium","gaps":["Direct binding of NOP14 to importins or p65 not shown","How a ribosome biogenesis factor controls nuclear import unclear"]},{"year":2021,"claim":"Extended the NRIP1/GSK-3β/β-catenin axis to colorectal cancer, linking NOP14 to GSK-3β phosphorylation-inactivation.","evidence":"Overexpression/knockdown, Western blot, migration and viability assays","pmids":["34218653"],"confidence":"Low","gaps":["No biochemical reconstitution of the pathway","Direction of Wnt effect differs from other contexts"]},{"year":2022,"claim":"Used pharmacological epistasis to place NOP14 upstream of Wnt/β-catenin in melanoma stem-like cells, affecting stemness.","evidence":"Lentiviral overexpression with BML-284 Wnt-activator rescue in A375/A875 cells","pmids":["35282769"],"confidence":"Low","gaps":["No direct biochemical mechanism linking NOP14 to Wnt components","Single lab epistasis only"]},{"year":2024,"claim":"Connected NOP14's ribosome role to growth signaling, showing it acts downstream of mTORC1-S6K to recruit mTORC2 to ER-bound ribosomes and stabilize Akt.","evidence":"Knockdown/overexpression, subcellular fractionation, Co-IP, and phospho-Akt Western blot in cancer cell lines","pmids":["38272224"],"confidence":"Medium","gaps":["Direct NOP14–mTORC2 contacts not mapped","Whether effect requires intact 40S biogenesis untested"]},{"year":2026,"claim":"Showed NOP14 activates Wnt/β-catenin and ribosome biogenesis to drive endothelial proliferation and angiogenesis in diabetic retinopathy.","evidence":"Knockdown/overexpression, Wnt reporter, in vivo PDR model and HG-treated HRECs","pmids":["41638389"],"confidence":"Low","gaps":["Wnt link inferred from protein levels without reconstitution","Opposite Wnt directionality versus melanoma context unexplained"]},{"year":null,"claim":"How a single nucleolar 40S biogenesis factor mechanistically controls disparate signaling outputs (NF-κB import, mTORC2 recruitment, mRNA stability, Wnt direction) remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No unifying biochemical mechanism distinguishing ribosome-dependent from ribosome-independent functions","Context-dependent direction of Wnt regulation unexplained","No structural model of NOP14 or its interaction interfaces"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[0]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[0]}],"complexes":[],"partners":["EMG1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P78316","full_name":"Nucleolar protein 14","aliases":["Nucleolar complex protein 14"],"length_aa":857,"mass_kda":97.7,"function":"Involved in nucleolar processing of pre-18S ribosomal RNA. Has a role in the nuclear export of 40S pre-ribosomal subunit to the cytoplasm (By similarity)","subcellular_location":"Nucleus, nucleolus","url":"https://www.uniprot.org/uniprotkb/P78316/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/NOP14","classification":"Common Essential","n_dependent_lines":1184,"n_total_lines":1208,"dependency_fraction":0.9801324503311258},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"BYSL","stoichiometry":4.0},{"gene":"EMG1","stoichiometry":4.0},{"gene":"IPO5","stoichiometry":4.0},{"gene":"BUD23","stoichiometry":0.2},{"gene":"CSNK2B","stoichiometry":0.2},{"gene":"DHX37","stoichiometry":0.2},{"gene":"LTV1","stoichiometry":0.2},{"gene":"RAB11A","stoichiometry":0.2},{"gene":"RPS16","stoichiometry":0.2},{"gene":"SEC31A","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NOP14","total_profiled":1310},"omim":[{"mim_id":"611531","title":"EMG1 N1-SPECIFIC PSEUDOURIDINE METHYLTRANSFERASE; EMG1","url":"https://www.omim.org/entry/611531"},{"mim_id":"611526","title":"NOP14 NUCLEOLAR PROTEIN; NOP14","url":"https://www.omim.org/entry/611526"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoli","reliability":"Enhanced"},{"location":"Nucleoplasm","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NOP14"},"hgnc":{"alias_symbol":["RES4-25","UTP2"],"prev_symbol":["C4orf9","NOL14"]},"alphafold":{"accession":"P78316","domains":[{"cath_id":"-","chopping":"582-750","consensus_level":"medium","plddt":81.9226,"start":582,"end":750}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P78316","model_url":"https://alphafold.ebi.ac.uk/files/AF-P78316-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P78316-F1-predicted_aligned_error_v6.png","plddt_mean":73.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NOP14","jax_strain_url":"https://www.jax.org/strain/search?query=NOP14"},"sequence":{"accession":"P78316","fasta_url":"https://rest.uniprot.org/uniprotkb/P78316.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P78316/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P78316"}},"corpus_meta":[{"pmid":"11694595","id":"PMC_11694595","title":"Novel stress-responsive genes EMG1 and NOP14 encode conserved, interacting proteins required for 40S ribosome biogenesis.","date":"2001","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/11694595","citation_count":89,"is_preprint":false},{"pmid":"31971654","id":"PMC_31971654","title":"CCND1, NOP14 and DNMT3B are involved in miR-502-5p-mediated inhibition of cell migration and proliferation in bladder cancer.","date":"2020","source":"Cell proliferation","url":"https://pubmed.ncbi.nlm.nih.gov/31971654","citation_count":46,"is_preprint":false},{"pmid":"28280038","id":"PMC_28280038","title":"Pancreatic Cancer Progression Relies upon Mutant p53-Induced Oncogenic Signaling Mediated by NOP14.","date":"2017","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28280038","citation_count":38,"is_preprint":false},{"pmid":"26213846","id":"PMC_26213846","title":"NOP14 suppresses breast cancer progression by inhibiting NRIP1/Wnt/β-catenin pathway.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26213846","citation_count":27,"is_preprint":false},{"pmid":"22425761","id":"PMC_22425761","title":"NOP14 promotes proliferation and metastasis of pancreatic cancer cells.","date":"2012","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/22425761","citation_count":24,"is_preprint":false},{"pmid":"30484495","id":"PMC_30484495","title":"NOP14 inhibits melanoma proliferation and metastasis by regulating Wnt/β-catenin signaling pathway.","date":"2018","source":"Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas","url":"https://pubmed.ncbi.nlm.nih.gov/30484495","citation_count":20,"is_preprint":false},{"pmid":"29440706","id":"PMC_29440706","title":"A homozygous NOP14 variant is likely to cause recurrent pregnancy loss.","date":"2018","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29440706","citation_count":13,"is_preprint":false},{"pmid":"34218653","id":"PMC_34218653","title":"NOP14 regulates the growth, migration, and invasion of colorectal cancer cells by modulating the NRIP1/GSK-3β/β-catenin signaling pathway.","date":"2021","source":"European journal of histochemistry : EJH","url":"https://pubmed.ncbi.nlm.nih.gov/34218653","citation_count":12,"is_preprint":false},{"pmid":"30514686","id":"PMC_30514686","title":"[miR-122-5p inhibits the proliferation of melanoma cells by targeting NOP14].","date":"2018","source":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","url":"https://pubmed.ncbi.nlm.nih.gov/30514686","citation_count":12,"is_preprint":false},{"pmid":"34607110","id":"PMC_34607110","title":"The natural product vioprolide A exerts anti-inflammatory actions through inhibition of its cellular target NOP14 and downregulation of importin-dependent NF-ĸB p65 nuclear translocation.","date":"2021","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/34607110","citation_count":11,"is_preprint":false},{"pmid":"35282769","id":"PMC_35282769","title":"The NOP14 nucleolar protein suppresses the function and stemness of melanoma stem-like cells through Wnt/beta-catenin signaling inactivation.","date":"2022","source":"Bioengineered","url":"https://pubmed.ncbi.nlm.nih.gov/35282769","citation_count":10,"is_preprint":false},{"pmid":"33814931","id":"PMC_33814931","title":"LncRNA NOP14-AS1 Promotes Tongue Squamous Cell Carcinoma Progression by Targeting MicroRNA-665/HMGB3 Axis.","date":"2021","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/33814931","citation_count":8,"is_preprint":false},{"pmid":"38272224","id":"PMC_38272224","title":"NOP14-mediated ribosome biogenesis is required for mTORC2 activation and predicts rapamycin sensitivity.","date":"2024","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/38272224","citation_count":6,"is_preprint":false},{"pmid":"35117729","id":"PMC_35117729","title":"EMG1 interacts with NOP14 to regulate the growth, migration, and invasion of melanoma cells via the Wnt/β-catenin pathway.","date":"2020","source":"Translational cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/35117729","citation_count":6,"is_preprint":false},{"pmid":"35665668","id":"PMC_35665668","title":"The protein biosynthesis inhibitor vioprolide A evokes anti-angiogenic and pro-survival actions by targeting NOP14 and decreasing VEGF receptor 2- and TAZ-signaling.","date":"2022","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/35665668","citation_count":5,"is_preprint":false},{"pmid":"39862153","id":"PMC_39862153","title":"Clinical diagnostic value and potential regulatory mechanisms of lncRNA NOP14-AS1 in chronic kidney disease.","date":"2025","source":"Nucleosides, nucleotides & nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/39862153","citation_count":2,"is_preprint":false},{"pmid":"40180667","id":"PMC_40180667","title":"Blocking lncRNA NOP14-AS1 overcomes 5-Fu resistance of colon cancer cells by modulating miR-30a-5p-LDHA-glucose metabolism pathway.","date":"2025","source":"Discover oncology","url":"https://pubmed.ncbi.nlm.nih.gov/40180667","citation_count":2,"is_preprint":false},{"pmid":"41638389","id":"PMC_41638389","title":"NOP14 promotes proliferative diabetic retinopathy through ribosome biogenesis and endothelial dysfunction via Wnt/β-catenin signaling activation.","date":"2026","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/41638389","citation_count":0,"is_preprint":false},{"pmid":"35137856","id":"PMC_35137856","title":"Erratum notice for: \"NOP14 inhibits melanoma proliferation and metastasis by regulating Wnt/β-catenin signaling pathway\" [Braz J Med Biol Res 2019;52(1):7952].","date":"2022","source":"Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas","url":"https://pubmed.ncbi.nlm.nih.gov/35137856","citation_count":0,"is_preprint":false},{"pmid":"36148317","id":"PMC_36148317","title":"LncRNA NOP14-AS1 Promotes Tongue Squamous Cell Carcinoma Progression by Targeting MicroRNA-665/HMGB3 Axis [Retraction].","date":"2022","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/36148317","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11292,"output_tokens":2500,"usd":0.035688,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9692,"output_tokens":2917,"usd":0.060693,"stage2_stop_reason":"end_turn"},"total_usd":0.096381,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"NOP14 is a nucleolar protein required for 40S ribosome biogenesis; yeast depleted of Nop14 show selectively reduced levels of 20S pre-rRNA and mature 18S rRNA and diminished cellular levels of the 40S ribosomal subunit.\",\n      \"method\": \"Yeast depletion experiments, Northern blot analysis of pre-rRNA processing intermediates\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean loss-of-function with specific pre-rRNA processing phenotype, replicated in mammalian ortholog context, multiple orthogonal methods\",\n      \"pmids\": [\"11694595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"EMG1 nuclear localization depends on its physical interaction with NOP14; the two proteins interact directly, and this interaction is required for 18S rRNA maturation and 40S ribosome production.\",\n      \"method\": \"Co-immunoprecipitation, subcellular fractionation/localization, genetic depletion in yeast\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction established with localization consequence, multiple orthogonal methods (Co-IP + localization + rRNA processing assay)\",\n      \"pmids\": [\"11694595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NOP14 increases the stability of mutant p53 (mutp53) mRNA, and the NOP14/mutp53 axis suppresses p21 expression at both transcriptional and post-transcriptional levels via induction of miR-17-5p in pancreatic cancer cells.\",\n      \"method\": \"NOP14 knockdown/overexpression, mRNA stability assay, Western blot, in vivo tumor mouse models (subcutaneous, orthotopic, intravenous injection)\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple cellular assays and in vivo models in a single lab; mRNA stability mechanistic link established but not independently replicated\",\n      \"pmids\": [\"28280038\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NOP14 enhances ERα expression and inhibits the Wnt/β-catenin pathway by up-regulating NRIP1 expression in breast cancer cells.\",\n      \"method\": \"NOP14 overexpression/knockdown, Western blot, in vivo and in vitro tumor assays\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement via OE/KD with protein level readouts but no direct mechanistic assay linking NOP14 to NRIP1 regulation\",\n      \"pmids\": [\"26213846\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NOP14 promotes colorectal cancer cell growth, migration, and invasion by activating NRIP1 expression and promoting phosphorylation-inactivation of GSK-3β, leading to upregulation of β-catenin (NRIP1/GSK-3β/β-catenin signaling pathway).\",\n      \"method\": \"GFP-NOP14 overexpression, siRNA knockdown, Western blot, Transwell migration assay, CCK-8, flow cytometry\",\n      \"journal\": \"European journal of histochemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single set of methods, pathway placement by OE/KD without direct biochemical reconstitution\",\n      \"pmids\": [\"34218653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Inhibition of NOP14 (via vioprolide A treatment or NOP14 knockdown) downregulates importin-dependent NF-κB p65 nuclear translocation, reducing NF-κB promoter activity and inflammatory gene expression in endothelial cells; NOP14 knockdown confirmed a causal link between NOP14 and the anti-inflammatory action of vioprolide A.\",\n      \"method\": \"NOP14 knockdown (siRNA), vioprolide A pharmacological inhibition, NF-κB reporter assay, nuclear translocation imaging, in vivo leukocyte trafficking (cremaster muscle) and choroidal neovascularization mouse models\",\n      \"journal\": \"Biomedicine & pharmacotherapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological and genetic approaches converge on same mechanism, in vivo and in vitro validation, single lab\",\n      \"pmids\": [\"34607110\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NOP14, as a 40S ribosome biogenesis factor and target of the mTORC1-S6K axis, is required for mTORC2 activation and Akt stabilization; the mTORC2 complex is recruited to the rough endoplasmic reticulum through association with ER-bound ribosomes, and NOP14 knockdown leads to mTORC2 inactivation and Akt destabilization.\",\n      \"method\": \"NOP14 knockdown/overexpression, subcellular fractionation, co-immunoprecipitation, Western blot (phospho-Akt), cancer cell lines\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fractionation and Co-IP establish spatial mechanism, multiple orthogonal methods in single lab; functional consequence (Akt destabilization) confirmed with loss-of-function\",\n      \"pmids\": [\"38272224\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EMG1 physically interacts with NOP14 (confirmed by GST pulldown and co-immunoprecipitation), and co-overexpression of EMG1 and NOP14 cooperatively decreases levels of WNT3a, β-catenin, phospho-GSK-3β, and c-Myc in melanoma cells.\",\n      \"method\": \"GST pulldown, co-immunoprecipitation, overexpression in melanoma cell lines, Western blot\",\n      \"journal\": \"Translational cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal binding confirmed by two orthogonal pulldown methods; functional consequence in cells; single lab\",\n      \"pmids\": [\"35117729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NOP14 overexpression inactivates Wnt/β-catenin signaling in melanoma stem-like cells (CD133+), reducing stemness markers (Nanog, SOX2, OCT4), colony-forming ability, and the proportion of CD133+ cells; the Wnt activator BML-284 rescued these effects, placing NOP14 upstream of Wnt/β-catenin in this context.\",\n      \"method\": \"NOP14 overexpression (lentiviral), Wnt activator rescue (BML-284), flow cytometry, colony formation assay, Western blot in A375 and A875 melanoma cells\",\n      \"journal\": \"Bioengineered\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — epistasis via pharmacological rescue, single lab, no direct biochemical mechanism linking NOP14 to Wnt components\",\n      \"pmids\": [\"35282769\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"NOP14 activates Wnt/β-catenin signaling in retinal endothelial cells (increased p-GSK-3β, β-catenin, Cyclin D1), promoting ribosome biogenesis and endothelial proliferation; NOP14 knockdown suppresses these effects and reduces angiogenesis-related proteins (CD31, VEGFA, PDGF, ANG2) in a PDR mouse model.\",\n      \"method\": \"NOP14 knockdown/overexpression, Wnt/β-catenin reporter assay, Western blot, in vivo PDR mouse model, in vitro HG-treated HRECs\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, mechanistic link to Wnt pathway inferred from protein level changes without direct biochemical reconstitution\",\n      \"pmids\": [\"41638389\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NOP14 is an evolutionarily conserved nucleolar protein that functions as a 40S ribosome biogenesis factor by interacting with EMG1 (required for EMG1 nuclear localization) and is required for 18S rRNA maturation; beyond ribosome assembly, NOP14 acts downstream of mTORC1-S6K signaling to support mTORC2 activation and Akt stability by facilitating recruitment of mTORC2 to ER-bound ribosomes, modulates NF-κB p65 nuclear translocation through importin-dependent mechanisms, stabilizes mutant p53 mRNA to drive oncogenic signaling, and regulates the Wnt/β-catenin pathway (through NRIP1/GSK-3β) in multiple cancer contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NOP14 is an evolutionarily conserved nucleolar ribosome biogenesis factor required for production of the small (40S) ribosomal subunit, with depletion selectively impairing processing of 20S pre-rRNA to mature 18S rRNA [#0]. It executes this function through a direct physical interaction with EMG1, which it retains in the nucleus; loss of this NOP14–EMG1 association blocks 18S rRNA maturation and 40S subunit assembly [#1, #7]. Beyond its core role in ribosome assembly, NOP14 couples ribosome biogenesis to growth signaling: acting downstream of the mTORC1–S6K axis, it is required for recruitment of mTORC2 to ER-bound ribosomes, supporting mTORC2 activation and Akt stabilization [#6]. NOP14 also governs importin-dependent nuclear translocation of NF-κB p65, controlling inflammatory gene expression in endothelial cells [#5], and stabilizes mutant p53 mRNA to suppress p21 via miR-17-5p induction in pancreatic cancer [#2]. Across multiple cancer and vascular contexts NOP14 modulates Wnt/β-catenin signaling through NRIP1 and GSK-3β, though the direction of this effect is context-dependent [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established NOP14's core cellular function by showing it is a nucleolar factor essential for the small ribosomal subunit, answering what process the protein serves.\",\n      \"evidence\": \"Yeast depletion with Northern analysis of pre-rRNA processing intermediates and 40S subunit levels\",\n      \"pmids\": [\"11694595\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not define the molecular activity of NOP14 within the processing machinery\", \"Structural basis for pre-rRNA processing role not resolved\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Identified EMG1 as a direct NOP14 partner whose nuclear localization depends on NOP14, linking the interaction mechanistically to 18S maturation.\",\n      \"evidence\": \"Co-immunoprecipitation, subcellular fractionation, and genetic depletion in yeast\",\n      \"pmids\": [\"11694595\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Interaction interface not mapped\", \"Whether other 40S assembly factors are recruited by NOP14 unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"First placed NOP14 in a cancer signaling context, linking it to ERα and Wnt/β-catenin via NRIP1 upregulation.\",\n      \"evidence\": \"Overexpression/knockdown with Western blot and tumor assays in breast cancer cells\",\n      \"pmids\": [\"26213846\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct mechanistic assay connecting NOP14 to NRIP1 regulation\", \"Single lab, protein-level readouts only\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Revealed a non-ribosomal RNA-stability function, showing NOP14 stabilizes mutant p53 mRNA to drive a p21-suppressing miR-17-5p axis.\",\n      \"evidence\": \"Knockdown/overexpression, mRNA stability assays, and in vivo pancreatic cancer models\",\n      \"pmids\": [\"28280038\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct RNA-binding by NOP14 not demonstrated\", \"Not independently replicated\", \"Mechanism of mRNA stabilization unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Confirmed the NOP14–EMG1 interaction biochemically in human cells and tied co-expression to suppression of Wnt/β-catenin signaling components.\",\n      \"evidence\": \"GST pulldown, co-immunoprecipitation, and overexpression in melanoma cells\",\n      \"pmids\": [\"35117729\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional link to Wnt is correlative protein-level\", \"Whether ribosome biogenesis role mediates the Wnt effect untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined NOP14 as a controller of importin-dependent NF-κB p65 nuclear translocation and the target of the anti-inflammatory compound vioprolide A.\",\n      \"evidence\": \"siRNA knockdown, pharmacological inhibition, NF-κB reporter and translocation imaging, and in vivo models\",\n      \"pmids\": [\"34607110\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding of NOP14 to importins or p65 not shown\", \"How a ribosome biogenesis factor controls nuclear import unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended the NRIP1/GSK-3β/β-catenin axis to colorectal cancer, linking NOP14 to GSK-3β phosphorylation-inactivation.\",\n      \"evidence\": \"Overexpression/knockdown, Western blot, migration and viability assays\",\n      \"pmids\": [\"34218653\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No biochemical reconstitution of the pathway\", \"Direction of Wnt effect differs from other contexts\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Used pharmacological epistasis to place NOP14 upstream of Wnt/β-catenin in melanoma stem-like cells, affecting stemness.\",\n      \"evidence\": \"Lentiviral overexpression with BML-284 Wnt-activator rescue in A375/A875 cells\",\n      \"pmids\": [\"35282769\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct biochemical mechanism linking NOP14 to Wnt components\", \"Single lab epistasis only\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected NOP14's ribosome role to growth signaling, showing it acts downstream of mTORC1-S6K to recruit mTORC2 to ER-bound ribosomes and stabilize Akt.\",\n      \"evidence\": \"Knockdown/overexpression, subcellular fractionation, Co-IP, and phospho-Akt Western blot in cancer cell lines\",\n      \"pmids\": [\"38272224\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct NOP14–mTORC2 contacts not mapped\", \"Whether effect requires intact 40S biogenesis untested\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed NOP14 activates Wnt/β-catenin and ribosome biogenesis to drive endothelial proliferation and angiogenesis in diabetic retinopathy.\",\n      \"evidence\": \"Knockdown/overexpression, Wnt reporter, in vivo PDR model and HG-treated HRECs\",\n      \"pmids\": [\"41638389\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Wnt link inferred from protein levels without reconstitution\", \"Opposite Wnt directionality versus melanoma context unexplained\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single nucleolar 40S biogenesis factor mechanistically controls disparate signaling outputs (NF-κB import, mTORC2 recruitment, mRNA stability, Wnt direction) remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying biochemical mechanism distinguishing ribosome-dependent from ribosome-independent functions\", \"Context-dependent direction of Wnt regulation unexplained\", \"No structural model of NOP14 or its interaction interfaces\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"EMG1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":5,"faith_pct":80.0}}