{"gene":"KANK3","run_date":"2026-06-10T01:55:23","timeline":{"discoveries":[{"year":2007,"finding":"KANK3 (as a member of the Kank family) contains conserved ankyrin-repeat, coiled-coil, and KN motif domains; overexpression of KANK3 in NIH3T3 cells promotes formation of actin stress fibers, suggesting a role in actin cytoskeleton organization.","method":"Overexpression in NIH3T3 cells, Western blotting, immunostaining, RT-PCR","journal":"Biochimica et biophysica acta","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single overexpression method, no mutagenesis or mechanistic dissection of how KANK3 promotes stress fibers","pmids":["17996375"],"is_preprint":false},{"year":2012,"finding":"NBP (zebrafish ortholog of human KANK3/Kank) interacts with the PTB domain of Numb adaptor protein via a conserved NGGY-containing region; genetic interaction between NBP and Numb was demonstrated by enhanced phenotypic defects in double morphants, and NBP localizes to sites of cell-cell contact and basal poles of differentiated cells, implicating it in cell adhesion and tissue integrity during epidermal and neurulation development.","method":"Yeast two-hybrid, morpholino knockdown (morphants), double morphant epistasis, protein localization by immunostaining","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast-two-hybrid binding assay plus genetic epistasis via double morphants and localization data, single lab","pmids":["22387208"],"is_preprint":false},{"year":2015,"finding":"KANK3 is a hypoxia-inducible, p53-dependent proapoptotic transcriptional target; its induction requires the DNA-binding and transactivation domains of p53 but not acetylation sites K120/K164 (which are required for DNA-damage-induced p53 apoptosis), placing KANK3 specifically in the hypoxia-p53 apoptotic pathway.","method":"RNA-seq of multiple cell lines under hypoxia, p53 domain mutant analysis, RT-PCR validation","journal":"The Journal of clinical investigation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple cell lines, p53 domain mutant dissection, but KANK3-specific mechanistic follow-up (e.g., direct rescue) not reported for KANK3 itself","pmids":["25961455"],"is_preprint":false},{"year":2017,"finding":"KANK3 is a substrate for the oxygen-sensor HIF1AN (hypoxia-inducible factor 1-alpha inhibitor/Factor Inhibiting HIF): HIF1AN hydroxylates KANK3 at three asparagine residues within its ankyrin repeat domain, as demonstrated by in vitro hydroxylation assay and mass spectrometry. KANK3 knockdown in hepatocellular carcinoma cells enhanced migration and invasion, while overexpression inhibited these behaviors; these effects were not observed under hypoxic conditions, indicating oxygen-dependent activity.","method":"In vitro hydroxylation assay, mass spectrometry (identification of hydroxylated asparagine residues), siRNA knockdown, overexpression, migration/invasion assays (Transwell), hypoxic vs. normoxic conditions","journal":"Cell biology international","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical hydroxylation assay with MS identification of specific residues plus functional cell-based loss-of-function/gain-of-function experiments, single lab but multiple orthogonal methods","pmids":["29047187"],"is_preprint":false},{"year":2020,"finding":"KANK3 localizes exclusively to vascular and lymphatic endothelial cells in mouse tissues, as determined by tissue distribution analysis, distinguishing it from other KANK family members and suggesting cell-type-specific function.","method":"Immunostaining and mRNA expression analysis across mouse tissues","journal":"Experimental cell research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization determined by immunostaining, single lab, no direct functional consequence demonstrated for KANK3 specifically","pmids":["33253712"],"is_preprint":false},{"year":2022,"finding":"KANK3 regulates proliferation, invasion, and migration of lung adenocarcinoma cells through the p38 MAPK signaling pathway; KANK3 overexpression increased p-p38 levels and inhibited cell growth and metastatic behaviors, while KANK3 silencing had opposite effects.","method":"Western blot (p38, p-p38), MTT proliferation assay, Transwell invasion, wound-healing migration assay, GSEA pathway enrichment, siRNA knockdown and overexpression in NCI-H1975 and PC-9 cells","journal":"Tissue & cell","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple orthogonal functional assays (proliferation, invasion, migration) plus pathway readout by Western blot, single lab, no rescue or mutagenesis","pmids":["36463587"],"is_preprint":false}],"current_model":"KANK3 is an ankyrin repeat domain-containing protein that functions as a hypoxia-inducible, p53-dependent proapoptotic target; it is hydroxylated at three asparagine residues within its ankyrin repeat domain by the oxygen sensor HIF1AN in a manner that confers oxygen-dependent activity, and it suppresses cancer cell migration, invasion, and proliferation at least in part through the p38 MAPK pathway, while also interacting (via its zebrafish ortholog) with the Numb PTB domain to regulate cell adhesion and tissue integrity."},"narrative":{"mechanistic_narrative":"KANK3 is an ankyrin-repeat, coiled-coil, and KN-motif domain protein implicated in actin cytoskeleton organization and the suppression of cancer cell migration, invasion, and proliferation [PMID:17996375, PMID:36463587]. It is a substrate of the oxygen sensor HIF1AN (Factor Inhibiting HIF), which hydroxylates KANK3 at three asparagine residues within its ankyrin repeat domain, and its tumor-suppressive activity in migration and invasion is oxygen-dependent, being lost under hypoxic conditions [PMID:29047187]. KANK3 restrains proliferation and metastatic behavior at least in part through activation of the p38 MAPK pathway, with overexpression elevating p-p38 levels and inhibiting cell growth [PMID:36463587]. KANK3 is also a hypoxia-inducible, p53-dependent proapoptotic transcriptional target, requiring the DNA-binding and transactivation domains of p53 and placing it specifically in the hypoxia-p53 apoptotic axis rather than the DNA-damage response [PMID:25961455]. Its zebrafish ortholog binds the Numb PTB domain via a conserved NGGY-containing region and acts at sites of cell-cell contact to regulate adhesion and tissue integrity during development [PMID:22387208].","teleology":[{"year":2007,"claim":"Established KANK3 as a domain-conserved Kank family member with a candidate role in actin organization, the first functional hint for an otherwise uncharacterized protein.","evidence":"Overexpression in NIH3T3 cells with immunostaining for stress fibers","pmids":["17996375"],"confidence":"Low","gaps":["Single overexpression assay with no mutagenesis","No direct binding partner or mechanism linking KANK3 to actin remodeling","Endogenous function not addressed"]},{"year":2012,"claim":"Identified a direct physical and genetic partner for the KANK3 ortholog, linking it to the Numb adaptor and cell-cell adhesion during development.","evidence":"Yeast two-hybrid, morpholino double-morphant epistasis, and localization in zebrafish","pmids":["22387208"],"confidence":"Medium","gaps":["Demonstrated for the zebrafish ortholog NBP, not directly for human KANK3","Functional significance of the Numb interaction in mammalian cells unknown","No structural mapping beyond the NGGY region"]},{"year":2015,"claim":"Placed KANK3 in a specific signaling context as a hypoxia-inducible, p53-dependent proapoptotic target distinct from the DNA-damage p53 program.","evidence":"RNA-seq under hypoxia across cell lines with p53 domain-mutant dissection and RT-PCR validation","pmids":["25961455"],"confidence":"Medium","gaps":["KANK3-specific rescue of the apoptotic phenotype not demonstrated","Direct p53 binding to KANK3 promoter not mapped","Mechanism by which KANK3 promotes apoptosis unresolved"]},{"year":2017,"claim":"Defined a biochemical oxygen-sensing mechanism by showing KANK3 is hydroxylated by HIF1AN at three ankyrin-domain asparagines, coupling its tumor-suppressive activity to oxygen availability.","evidence":"In vitro hydroxylation assay with MS site identification plus knockdown/overexpression migration-invasion assays under normoxia vs hypoxia in HCC cells","pmids":["29047187"],"confidence":"High","gaps":["Functional consequence of each hydroxylation site not individually tested by mutagenesis","Downstream effector of hydroxylated KANK3 unknown","Link between hydroxylation and the p38 or p53 pathways unexplored"]},{"year":2020,"claim":"Refined KANK3 expression to vascular and lymphatic endothelial cells, distinguishing it from other Kank members and hinting at a cell-type-restricted role.","evidence":"Immunostaining and mRNA analysis across mouse tissues","pmids":["33253712"],"confidence":"Low","gaps":["Localization only, no functional consequence demonstrated","Endothelial-specific function untested","Relationship to its tumor cell roles unclear"]},{"year":2022,"claim":"Connected KANK3's anti-tumor activity to a defined signaling axis by showing it suppresses lung adenocarcinoma proliferation, invasion, and migration via p38 MAPK.","evidence":"siRNA/overexpression with proliferation, Transwell, wound-healing assays and p-p38 Western blot in NCI-H1975 and PC-9 cells","pmids":["36463587"],"confidence":"Medium","gaps":["No rescue or mutagenesis to confirm p38 dependence","Mechanism by which KANK3 activates p38 not defined","Relationship to HIF1AN hydroxylation and p53 induction not integrated"]},{"year":null,"claim":"How the oxygen-dependent HIF1AN hydroxylation, p53-dependent induction, and p38 MAPK signaling integrate into a single coherent KANK3 mechanism remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No unifying model linking hydroxylation, transcriptional induction, and p38 output","No structural model of the ankyrin domain bound to HIF1AN or Numb in human cells","Endothelial versus tumor-cell function not reconciled"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[5]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[5]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[2,3]}],"complexes":[],"partners":["HIF1AN","NUMB","TP53"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q6NY19","full_name":"KN motif and ankyrin repeat domain-containing protein 3","aliases":["Ankyrin repeat domain-containing protein 47"],"length_aa":821,"mass_kda":85.9,"function":"May be involved in the control of cytoskeleton formation by regulating actin polymerization","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q6NY19/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/KANK3","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/KANK3","total_profiled":1310},"omim":[{"mim_id":"614612","title":"KN MOTIF- AND ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 4; KANK4","url":"https://www.omim.org/entry/614612"},{"mim_id":"614611","title":"KN MOTIF- AND ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 3; KANK3","url":"https://www.omim.org/entry/614611"},{"mim_id":"614610","title":"KN MOTIF- AND ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 2; KANK2","url":"https://www.omim.org/entry/614610"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/KANK3"},"hgnc":{"alias_symbol":["FLJ46061"],"prev_symbol":["ANKRD47"]},"alphafold":{"accession":"Q6NY19","domains":[{"cath_id":"1.25.40.20","chopping":"551-722","consensus_level":"medium","plddt":92.7375,"start":551,"end":722}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6NY19","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q6NY19-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q6NY19-F1-predicted_aligned_error_v6.png","plddt_mean":64.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=KANK3","jax_strain_url":"https://www.jax.org/strain/search?query=KANK3"},"sequence":{"accession":"Q6NY19","fasta_url":"https://rest.uniprot.org/uniprotkb/Q6NY19.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q6NY19/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6NY19"}},"corpus_meta":[{"pmid":"25961455","id":"PMC_25961455","title":"Hypoxia-induced p53 modulates both apoptosis and radiosensitivity via AKT.","date":"2015","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/25961455","citation_count":110,"is_preprint":false},{"pmid":"17996375","id":"PMC_17996375","title":"Kank proteins: a new family of ankyrin-repeat domain-containing proteins.","date":"2007","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/17996375","citation_count":43,"is_preprint":false},{"pmid":"27292017","id":"PMC_27292017","title":"Evolutionary and developmental analysis reveals KANK genes were co-opted for vertebrate vascular development.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27292017","citation_count":20,"is_preprint":false},{"pmid":"33309958","id":"PMC_33309958","title":"KANK family proteins in cancer.","date":"2020","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/33309958","citation_count":18,"is_preprint":false},{"pmid":"35065254","id":"PMC_35065254","title":"Identification of three immune subtypes characterized by distinct tumor immune microenvironment and therapeutic response in stomach adenocarcinoma.","date":"2022","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/35065254","citation_count":16,"is_preprint":false},{"pmid":"29047187","id":"PMC_29047187","title":"A novel HIF1AN substrate KANK3 plays a tumor-suppressive role in hepatocellular carcinoma.","date":"2017","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/29047187","citation_count":13,"is_preprint":false},{"pmid":"38460933","id":"PMC_38460933","title":"Transcriptomic and proteomic study of cancer cell lines exposed to actinomycin D and nutlin-3a reveals numerous, novel candidates for p53-regulated genes.","date":"2024","source":"Chemico-biological interactions","url":"https://pubmed.ncbi.nlm.nih.gov/38460933","citation_count":11,"is_preprint":false},{"pmid":"34764646","id":"PMC_34764646","title":"Immune Characteristics Analysis and Transcriptional Regulation Prediction Based on Gene Signatures of Chronic Obstructive Pulmonary Disease.","date":"2021","source":"International journal of chronic obstructive pulmonary disease","url":"https://pubmed.ncbi.nlm.nih.gov/34764646","citation_count":11,"is_preprint":false},{"pmid":"22387208","id":"PMC_22387208","title":"NBP, a zebrafish homolog of human Kank3, is a novel Numb interactor essential for epidermal integrity and neurulation.","date":"2012","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/22387208","citation_count":9,"is_preprint":false},{"pmid":"33253712","id":"PMC_33253712","title":"Tissue distribution and subcellular localization of the family of Kidney Ankyrin Repeat Domain (KANK) proteins.","date":"2020","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/33253712","citation_count":7,"is_preprint":false},{"pmid":"39340989","id":"PMC_39340989","title":"Spatial transcriptomics reveal tumor microenvironment and SLCO2A1 correlated with tumor suppression in hypopharyngeal squamous cell carcinoma.","date":"2024","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39340989","citation_count":5,"is_preprint":false},{"pmid":"36463587","id":"PMC_36463587","title":"KANK3 mediates the p38 MAPK pathway to regulate the proliferation and invasion of lung adenocarcinoma cells.","date":"2022","source":"Tissue & cell","url":"https://pubmed.ncbi.nlm.nih.gov/36463587","citation_count":3,"is_preprint":false},{"pmid":"39044124","id":"PMC_39044124","title":"Pterostilbene exerts anti-lung squamous cell carcinoma function by suppressing the level of KANK3.","date":"2024","source":"Chemical biology & drug design","url":"https://pubmed.ncbi.nlm.nih.gov/39044124","citation_count":2,"is_preprint":false},{"pmid":"39596293","id":"PMC_39596293","title":"Molecules That Have Rarely Been Studied in Lymphatic Endothelial Cells.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39596293","citation_count":2,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8982,"output_tokens":1613,"usd":0.02557,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8428,"output_tokens":2372,"usd":0.05072,"stage2_stop_reason":"end_turn"},"total_usd":0.07629,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"KANK3 (as a member of the Kank family) contains conserved ankyrin-repeat, coiled-coil, and KN motif domains; overexpression of KANK3 in NIH3T3 cells promotes formation of actin stress fibers, suggesting a role in actin cytoskeleton organization.\",\n      \"method\": \"Overexpression in NIH3T3 cells, Western blotting, immunostaining, RT-PCR\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single overexpression method, no mutagenesis or mechanistic dissection of how KANK3 promotes stress fibers\",\n      \"pmids\": [\"17996375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"NBP (zebrafish ortholog of human KANK3/Kank) interacts with the PTB domain of Numb adaptor protein via a conserved NGGY-containing region; genetic interaction between NBP and Numb was demonstrated by enhanced phenotypic defects in double morphants, and NBP localizes to sites of cell-cell contact and basal poles of differentiated cells, implicating it in cell adhesion and tissue integrity during epidermal and neurulation development.\",\n      \"method\": \"Yeast two-hybrid, morpholino knockdown (morphants), double morphant epistasis, protein localization by immunostaining\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast-two-hybrid binding assay plus genetic epistasis via double morphants and localization data, single lab\",\n      \"pmids\": [\"22387208\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"KANK3 is a hypoxia-inducible, p53-dependent proapoptotic transcriptional target; its induction requires the DNA-binding and transactivation domains of p53 but not acetylation sites K120/K164 (which are required for DNA-damage-induced p53 apoptosis), placing KANK3 specifically in the hypoxia-p53 apoptotic pathway.\",\n      \"method\": \"RNA-seq of multiple cell lines under hypoxia, p53 domain mutant analysis, RT-PCR validation\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple cell lines, p53 domain mutant dissection, but KANK3-specific mechanistic follow-up (e.g., direct rescue) not reported for KANK3 itself\",\n      \"pmids\": [\"25961455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"KANK3 is a substrate for the oxygen-sensor HIF1AN (hypoxia-inducible factor 1-alpha inhibitor/Factor Inhibiting HIF): HIF1AN hydroxylates KANK3 at three asparagine residues within its ankyrin repeat domain, as demonstrated by in vitro hydroxylation assay and mass spectrometry. KANK3 knockdown in hepatocellular carcinoma cells enhanced migration and invasion, while overexpression inhibited these behaviors; these effects were not observed under hypoxic conditions, indicating oxygen-dependent activity.\",\n      \"method\": \"In vitro hydroxylation assay, mass spectrometry (identification of hydroxylated asparagine residues), siRNA knockdown, overexpression, migration/invasion assays (Transwell), hypoxic vs. normoxic conditions\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical hydroxylation assay with MS identification of specific residues plus functional cell-based loss-of-function/gain-of-function experiments, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"29047187\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"KANK3 localizes exclusively to vascular and lymphatic endothelial cells in mouse tissues, as determined by tissue distribution analysis, distinguishing it from other KANK family members and suggesting cell-type-specific function.\",\n      \"method\": \"Immunostaining and mRNA expression analysis across mouse tissues\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization determined by immunostaining, single lab, no direct functional consequence demonstrated for KANK3 specifically\",\n      \"pmids\": [\"33253712\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"KANK3 regulates proliferation, invasion, and migration of lung adenocarcinoma cells through the p38 MAPK signaling pathway; KANK3 overexpression increased p-p38 levels and inhibited cell growth and metastatic behaviors, while KANK3 silencing had opposite effects.\",\n      \"method\": \"Western blot (p38, p-p38), MTT proliferation assay, Transwell invasion, wound-healing migration assay, GSEA pathway enrichment, siRNA knockdown and overexpression in NCI-H1975 and PC-9 cells\",\n      \"journal\": \"Tissue & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple orthogonal functional assays (proliferation, invasion, migration) plus pathway readout by Western blot, single lab, no rescue or mutagenesis\",\n      \"pmids\": [\"36463587\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KANK3 is an ankyrin repeat domain-containing protein that functions as a hypoxia-inducible, p53-dependent proapoptotic target; it is hydroxylated at three asparagine residues within its ankyrin repeat domain by the oxygen sensor HIF1AN in a manner that confers oxygen-dependent activity, and it suppresses cancer cell migration, invasion, and proliferation at least in part through the p38 MAPK pathway, while also interacting (via its zebrafish ortholog) with the Numb PTB domain to regulate cell adhesion and tissue integrity.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"KANK3 is an ankyrin-repeat, coiled-coil, and KN-motif domain protein implicated in actin cytoskeleton organization and the suppression of cancer cell migration, invasion, and proliferation [#0, #5]. It is a substrate of the oxygen sensor HIF1AN (Factor Inhibiting HIF), which hydroxylates KANK3 at three asparagine residues within its ankyrin repeat domain, and its tumor-suppressive activity in migration and invasion is oxygen-dependent, being lost under hypoxic conditions [#3]. KANK3 restrains proliferation and metastatic behavior at least in part through activation of the p38 MAPK pathway, with overexpression elevating p-p38 levels and inhibiting cell growth [#5]. KANK3 is also a hypoxia-inducible, p53-dependent proapoptotic transcriptional target, requiring the DNA-binding and transactivation domains of p53 and placing it specifically in the hypoxia-p53 apoptotic axis rather than the DNA-damage response [#2]. Its zebrafish ortholog binds the Numb PTB domain via a conserved NGGY-containing region and acts at sites of cell-cell contact to regulate adhesion and tissue integrity during development [#1].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established KANK3 as a domain-conserved Kank family member with a candidate role in actin organization, the first functional hint for an otherwise uncharacterized protein.\",\n      \"evidence\": \"Overexpression in NIH3T3 cells with immunostaining for stress fibers\",\n      \"pmids\": [\"17996375\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single overexpression assay with no mutagenesis\", \"No direct binding partner or mechanism linking KANK3 to actin remodeling\", \"Endogenous function not addressed\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identified a direct physical and genetic partner for the KANK3 ortholog, linking it to the Numb adaptor and cell-cell adhesion during development.\",\n      \"evidence\": \"Yeast two-hybrid, morpholino double-morphant epistasis, and localization in zebrafish\",\n      \"pmids\": [\"22387208\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Demonstrated for the zebrafish ortholog NBP, not directly for human KANK3\", \"Functional significance of the Numb interaction in mammalian cells unknown\", \"No structural mapping beyond the NGGY region\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Placed KANK3 in a specific signaling context as a hypoxia-inducible, p53-dependent proapoptotic target distinct from the DNA-damage p53 program.\",\n      \"evidence\": \"RNA-seq under hypoxia across cell lines with p53 domain-mutant dissection and RT-PCR validation\",\n      \"pmids\": [\"25961455\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"KANK3-specific rescue of the apoptotic phenotype not demonstrated\", \"Direct p53 binding to KANK3 promoter not mapped\", \"Mechanism by which KANK3 promotes apoptosis unresolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined a biochemical oxygen-sensing mechanism by showing KANK3 is hydroxylated by HIF1AN at three ankyrin-domain asparagines, coupling its tumor-suppressive activity to oxygen availability.\",\n      \"evidence\": \"In vitro hydroxylation assay with MS site identification plus knockdown/overexpression migration-invasion assays under normoxia vs hypoxia in HCC cells\",\n      \"pmids\": [\"29047187\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of each hydroxylation site not individually tested by mutagenesis\", \"Downstream effector of hydroxylated KANK3 unknown\", \"Link between hydroxylation and the p38 or p53 pathways unexplored\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Refined KANK3 expression to vascular and lymphatic endothelial cells, distinguishing it from other Kank members and hinting at a cell-type-restricted role.\",\n      \"evidence\": \"Immunostaining and mRNA analysis across mouse tissues\",\n      \"pmids\": [\"33253712\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Localization only, no functional consequence demonstrated\", \"Endothelial-specific function untested\", \"Relationship to its tumor cell roles unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Connected KANK3's anti-tumor activity to a defined signaling axis by showing it suppresses lung adenocarcinoma proliferation, invasion, and migration via p38 MAPK.\",\n      \"evidence\": \"siRNA/overexpression with proliferation, Transwell, wound-healing assays and p-p38 Western blot in NCI-H1975 and PC-9 cells\",\n      \"pmids\": [\"36463587\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No rescue or mutagenesis to confirm p38 dependence\", \"Mechanism by which KANK3 activates p38 not defined\", \"Relationship to HIF1AN hydroxylation and p53 induction not integrated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the oxygen-dependent HIF1AN hydroxylation, p53-dependent induction, and p38 MAPK signaling integrate into a single coherent KANK3 mechanism remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying model linking hydroxylation, transcriptional induction, and p38 output\", \"No structural model of the ankyrin domain bound to HIF1AN or Numb in human cells\", \"Endothelial versus tumor-cell function not reconciled\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"HIF1AN\", \"NUMB\", \"TP53\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}