{"gene":"TANK","run_date":"2026-06-10T10:51:54","timeline":{"discoveries":[{"year":1996,"finding":"TANK (I-TRAF) was identified as a novel intracellular protein that binds to the conserved TRAF-C domain of TRAF1, TRAF2, and TRAF3. Overexpression of I-TRAF inhibits TRAF2-mediated NF-κB activation downstream of CD40 and both TNF receptors, suggesting TANK acts as a natural regulator of TRAF function by maintaining TRAFs in a latent state.","method":"Yeast two-hybrid screening, co-immunoprecipitation, cotransfection NF-κB reporter assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — independently identified in two labs (Rothe et al. PMID:8710854 and Cheng & Baltimore PMID:8608943) using multiple orthogonal methods including yeast two-hybrid, Co-IP, and functional reporter assays","pmids":["8710854","8608943"],"is_preprint":false},{"year":1996,"finding":"TANK and TRAF2 synergistically activate NF-κB in cotransfection experiments. This synergistic activation requires both the amino-terminal portion of TANK and the RING finger domain of TRAF2. TANK has an internally inhibitory carboxyl terminus, and TRAF2 binding overcomes this internal inhibition.","method":"Cotransfection NF-κB luciferase reporter assays, deletion mutagenesis","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Moderate — functional reporter assays combined with domain mutagenesis in a focused mechanistic study, replicated by related findings in PMID:8710854","pmids":["8608943"],"is_preprint":false},{"year":1999,"finding":"TANK interacts with TBK1 (TANK-binding kinase 1), a novel IKK-related kinase. TBK1, TANK, and TRAF2 form a ternary complex, and complex formation is required for TBK1 kinase activity. Kinase-inactive TBK1 inhibits TANK-mediated NF-κB activation but does not block TNF-α, IL-1, or CD40 signaling, positioning the TBK1-TANK-TRAF2 complex upstream of NIK and the IKK complex.","method":"Co-immunoprecipitation, kinase-dead dominant-negative constructs, NF-κB reporter assays, genetic epistasis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP identifying the ternary complex, dominant-negative kinase assays, and epistasis experiments all in one study; foundational result replicated in subsequent work","pmids":["10581243"],"is_preprint":false},{"year":2000,"finding":"IKK-i (IKKε) phosphorylates TANK/I-TRAF in vitro in the middle portion of TANK that associates with TRAF2. Following IKK-i-mediated phosphorylation of TANK, TRAF2 is released from the TANK/TRAF2 complex. The N-terminal domain of TANK mediates its interaction with the C-terminal portion of IKK-i, and NF-κB activation by IKK-i is blocked by co-expression of the N-terminal domain of TANK.","method":"Yeast two-hybrid (IKK-i/TANK interaction), in vitro kinase assay, co-immunoprecipitation, dominant-negative overexpression","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro kinase assay with domain mapping plus Co-IP and functional assays, single lab","pmids":["10759890"],"is_preprint":false},{"year":2002,"finding":"Crystal structure of a TANK peptide bound to TRAF3 was determined, showing that TANK and CD40 bind to the same crevice on TRAF3. TANK presents a boomerang-like structure at the recognition motif PxQxT that is distinct from the hairpin loop of CD40. Mutagenesis confirmed critical TANK contact residues for binding to TRAF3 and TRAF2; TANK competes with CD40 for the TRAF binding site as demonstrated by isothermal titration calorimetry and competition assays.","method":"X-ray crystallography, site-directed mutagenesis, isothermal titration calorimetry, competition binding assays","journal":"Structure (London, England : 1993)","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus mutagenesis plus quantitative binding assays in one study","pmids":["12005438"],"is_preprint":false},{"year":2006,"finding":"TNF-α stimulation promotes TANK recruitment to the IKK complex via a newly characterized C-terminal zinc finger in TANK. IKKβ phosphorylates TANK upon TNF-α stimulation, and this phosphorylation negatively regulates TANK binding to NEMO. Reduced TANK expression (by RNAi) attenuates TNF-α-mediated induction of a subset of NF-κB target genes through decreased p65 transactivation potential.","method":"Co-immunoprecipitation, RNAi knockdown, NF-κB reporter assays, domain mapping","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for complex mapping plus functional RNAi knockdown, single lab with multiple methods","pmids":["16336209"],"is_preprint":false},{"year":2006,"finding":"TFG (TRK-fused gene) was identified as a novel TANK-interacting protein through yeast two-hybrid screening and confirmed by in vitro and in vivo co-immunoprecipitation. TFG and NEMO may be part of the same high molecular weight complex. TFG enhances TANK-induced NF-κB activation.","method":"Yeast two-hybrid, co-immunoprecipitation, NF-κB reporter assay","journal":"Journal of cellular physiology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP/yeast two-hybrid in one lab with limited mechanistic follow-up specific to TANK","pmids":["16547966"],"is_preprint":false},{"year":2007,"finding":"TANK was identified as a scaffold protein that assembles TBK1/IKKε complexes required for IRF3/7 phosphorylation in some (but not all) Toll-like receptor-dependent signaling pathways. TANK is part of a TRAF3-containing complex. Upon LPS stimulation, TANK is heavily phosphorylated by TBK1-IKKε and undergoes Lys63-linked polyubiquitination in a manner that does not require TBK1-IKKε kinase activity.","method":"Co-immunoprecipitation, in vitro kinase assay, RNAi knockdown, ubiquitin linkage analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, kinase assay, RNAi, ubiquitin linkage) in one focused study of TANK's scaffold function","pmids":["17823124"],"is_preprint":false},{"year":2007,"finding":"SINTBAD, NAP1, and TANK share a conserved TBK1/IKKi-binding domain (TBD) predicted to form an alpha-helix with residues essential for kinase binding on one face. Isolated TBDs from each adaptor compete with each other for binding to TBK1 and prevent poly(I:C)-induced IRF-dependent transcription, establishing TANK as one of multiple adaptors linking TBK1/IKKi to virus-activated signaling.","method":"Co-immunoprecipitation, competition assays, siRNA knockdown, IRF-dependent transcription reporter assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — TBD competition assays plus RNAi plus reporter assays; TANK's TBK1-binding domain structurally defined and functionally validated","pmids":["17568778"],"is_preprint":false},{"year":2009,"finding":"SARS coronavirus M protein physically associates with TRAF3, TBK1, IKKε, and RIG-I, and prevents formation of the TRAF3·TANK·TBK1/IKKε complex, thereby inhibiting TBK1/IKKε-dependent activation of IRF3/IRF7 and type I interferon production. M protein has no influence when IRF3 or IRF7 are overexpressed directly, indicating the block is upstream at complex assembly.","method":"Co-immunoprecipitation, IRF-dependent transcription reporter assay, overexpression studies","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating complex disruption plus functional reporter assays, single lab","pmids":["19380580"],"is_preprint":false},{"year":2010,"finding":"TANK directly interacts with PLK1 (polo-like kinase 1). PLK1, TANK, and NEMO (IKKγ) form a ternary complex in vivo. PLK1 negatively regulates TNF-induced IKK activation by inhibiting ubiquitination of NEMO, establishing TANK as a scaffold that recruits PLK1 to suppress NF-κB signaling.","method":"Co-immunoprecipitation, ubiquitination assay, NF-κB reporter assays, NF-κB DNA-binding assay","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP for ternary complex plus functional ubiquitination and reporter assays, single lab","pmids":["20484576"],"is_preprint":false},{"year":2010,"finding":"TANK participates in limiting non-canonical NF-κB signaling downstream of TACI. TANK deficiency (by siRNA) impairs TACI-dependent inhibition of NF-κB2 p100 processing. TANK inhibits TRAF2-mediated cIAP1 inactivation and thereby facilitates cIAP1-mediated ubiquitination of NIK to suppress non-canonical NF-κB signaling.","method":"siRNA knockdown, co-immunoprecipitation, ubiquitination assays, NF-κB reporter assays","journal":"Journal of receptor and signal transduction research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA loss-of-function with defined pathway phenotype plus Co-IP, single lab","pmids":["20394400"],"is_preprint":false},{"year":2011,"finding":"TANK is required for the activation of IKKε and for optimal activation of TBK1 in macrophages stimulated through both MyD88- and TRIF-dependent TLR pathways. TANK is required for the interaction between IKK-related kinases and canonical IKKs, enabling IKK-related kinases to negatively regulate canonical IKKs. In TANK−/− macrophages, IKKε activation is abolished, TBK1 activation is reduced, but IRF3 phosphorylation and IFNβ production remain normal.","method":"TANK knockout macrophages, kinase activity assays, co-immunoprecipitation, IRF3 phosphorylation assays, IFNβ ELISA","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic KO with multiple phenotypic readouts and Co-IP, separating TANK's roles in IKK cross-talk vs. IFN production","pmids":["21949249"],"is_preprint":false},{"year":2011,"finding":"SUMOylation of TANK at the evolutionarily conserved Lys282 is triggered by IKKε and TBK1 kinase activities. Stimulation of TLR7 induces TANK SUMOylation, which weakens the interaction between TANK and IKKε, thereby relieving TANK's negative function on TLR7 signal propagation. Reconstitution experiments show that absence of TANK SUMOylation impairs inducible expression of TLR7-dependent target genes.","method":"SUMOylation assays, site-directed mutagenesis (K282), co-immunoprecipitation, reconstitution in TANK-deficient cells, reporter assays","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis of the SUMO site plus Co-IP plus reconstitution experiments, multiple methods in one study","pmids":["21212807"],"is_preprint":false},{"year":2012,"finding":"TANK is a negative regulator of osteoclast differentiation. TANK expression is upregulated during RANKL-induced osteoclastogenesis. Tank−/− cells show increased osteoclastogenesis with increased TRAF6 ubiquitination and enhanced canonical NF-κB activation in response to RANKL. Tank−/− mice develop severe trabecular bone loss due to enhanced bone erosion.","method":"TANK knockout mouse model, osteoclast differentiation assays, ubiquitination assays, NF-κB activation assays, bone histomorphometry","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO with defined cellular phenotype, mechanistic ubiquitination assays, and in vivo bone phenotype","pmids":["22773835"],"is_preprint":false},{"year":2012,"finding":"In the context of virus-triggered signaling, TANK mono-ubiquitination (mediated by Pellino3 downstream of IRAK1/4 via SR-A1) inhibits TBK1 recruitment to TRAF3, thereby blocking TBK1-TRAF3 interaction and subsequent IRF3 activation and IFNβ expression. oxLDL stimulates this TANK mono-ubiquitination pathway.","method":"Co-immunoprecipitation, ubiquitination assays, RNAi knockdown, IRF3 activation assays","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP showing TBK1-TRAF3 disruption plus ubiquitination and RNAi assays, single lab","pmids":["22330071"],"is_preprint":false},{"year":2015,"finding":"TANK negatively regulates genotoxic NF-κB activation by facilitating USP10-dependent deubiquitination of TRAF6. TANK forms a complex with MCPIP1 (ZC3H12A) and the deubiquitinase USP10, which is essential for TRAF6 deubiquitination and resolution of DNA damage-induced NF-κB activation. CRISPR/Cas9 deletion of TANK in human cells enhances NF-κB activation after genotoxic treatment. The TANK-MCPIP1-USP10 complex also decreases TRAF6 ubiquitination downstream of IL-1β and LPS.","method":"CRISPR/Cas9 knockout, co-immunoprecipitation, ubiquitination assays, NF-κB reporter assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — CRISPR KO plus Co-IP identifying trimeric complex plus functional ubiquitination and NF-κB assays in multiple stimulation contexts","pmids":["25861989"],"is_preprint":false},{"year":2015,"finding":"Encephalomyocarditis virus (EMCV) 3C protease cleaves TANK at Q197 and Q291, dependent on 3C cysteine protease activity. Cleavage of TANK by EMCV 3C impairs TANK's ability to inhibit TRAF6-mediated NF-κB signaling. Several other picornavirus proteases (FMDV, PRRSV, EAV) also cleave TANK.","method":"In vitro protease cleavage assays, site-directed mutagenesis, NF-κB reporter assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro cleavage assay with mutagenesis of cleavage sites plus functional NF-κB assay, single lab","pmids":["26363073"],"is_preprint":false},{"year":2017,"finding":"Crystal structure of the TRAF1/TANK peptide complex was determined (PDB: 5H10). TANK binds TRAF1 using the minor minimal consensus motif Px(Q/E)xT. Quantitative interaction experiments showed TANK peptide interacts with TRAF1 and TRAF2 with similar micromolar affinity.","method":"X-ray crystallography, isothermal titration calorimetry, quantitative binding assays","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus quantitative binding measurements, single lab but Tier 1 method","pmids":["28155233"],"is_preprint":false},{"year":2017,"finding":"Seneca Valley virus 3C protease mediates cleavage of TANK (along with MAVS and TRIF) at specific sites requiring 3C protease activity. Cleaved TANK loses the ability to regulate PRR-mediated IFN production, and TANK cleavage facilitates TRAF6-induced NF-κB activation.","method":"In vitro protease cleavage assays, IFN production assays, NF-κB reporter assays","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro cleavage assay with functional IFN and NF-κB readouts, single lab","pmids":["28566380"],"is_preprint":false},{"year":2011,"finding":"In the context of TLR7 signaling, TANK SUMOylation (induced by TBK1/IKKε) weakens the TANK-IKKε interaction, thereby relieving TANK's constitutive negative function on downstream signal propagation. Separately, TANK expression is up-regulated in injured sensory neurons (DRG) and is transcriptionally regulated by the injury-associated transcription factor Sox11 via direct binding to two Sox motifs in the TANK 5'-UTR.","method":"Chromatin immunoprecipitation, luciferase reporter with Sox site mutagenesis, Sox11 siRNA knockdown, quantitative PCR","journal":"Neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus mutagenesis of promoter sites plus siRNA knockdown, single lab; establishes Sox11 as a transcriptional regulator of TANK","pmids":["23201825"],"is_preprint":false},{"year":2019,"finding":"TANK knockout mice display altered alcohol drinking behavior and enhanced anxiety-related behavior, and show increased NF-κB activation in the insular cortex following alcohol exposure, suggesting TANK regulates cortical NF-κB-dependent neuroimmune signaling in the context of aversive emotional processing.","method":"Tank knockout mouse model, behavioral assays, NF-κB activation assays in brain tissue","journal":"Cerebral cortex (New York, N.Y. : 1991)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with behavioral phenotype and biochemical NF-κB readout in brain tissue, single lab","pmids":["30721969"],"is_preprint":false},{"year":2022,"finding":"PLK1 co-localizes and physically interacts with TANK in intestinal epithelial (Caco-2) cells. TANK overexpression impairs the protective effect of PLK1 on LPS-induced intestinal epithelial injury and NF-κB activation, and TANK siRNA knockdown suppresses NF-κB signaling and ameliorates mitochondrial dysfunction and apoptosis induced by LPS. This defines a PLK1/TANK/NF-κB axis regulating intestinal barrier function.","method":"Co-immunoprecipitation, siRNA knockdown, NF-κB signaling assays, mitochondrial dynamics assays, apoptosis assays, in vivo CLP model","journal":"Molecular medicine (Cambridge, Mass.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus siRNA plus in vivo model, single lab with multiple methods","pmids":["36581806"],"is_preprint":false}],"current_model":"TANK (I-TRAF) is a scaffold/adaptor protein that binds to the TRAF-C domains of TRAF1, TRAF2, and TRAF3 via a PxQxT-like motif, forms a ternary complex with TBK1 and TRAF2 that is required for TBK1 kinase activity and NF-κB activation, and assembles the TRAF3·TANK·TBK1/IKKε complex essential for IRF3/7 phosphorylation and type I interferon induction; TANK also acts as a negative regulator of NF-κB by recruiting PLK1 to inhibit NEMO ubiquitination and by facilitating USP10-dependent deubiquitination of TRAF6, and its activity is fine-tuned by IKKε/TBK1-induced phosphorylation and SUMOylation at K282, the latter relieving its inhibitory function during TLR7 signaling."},"narrative":{"mechanistic_narrative":"TANK (I-TRAF) is an intracellular scaffold/adaptor protein that integrates TRAF-family signaling to both activate and restrain NF-κB and type I interferon induction [PMID:8710854, PMID:8608943, PMID:10581243]. It engages the conserved TRAF-C domains of TRAF1, TRAF2, and TRAF3 through a boomerang-shaped PxQxT-like recognition motif that occupies the same TRAF crevice used by receptor tails such as CD40, allowing TANK to compete for TRAF binding [PMID:8710854, PMID:8608943, PMID:12005438, PMID:28155233]. Through an N-terminal region TANK nucleates a ternary complex with the IKK-related kinase TBK1 and TRAF2 that is required for TBK1 kinase activity, and a shared TBK1/IKKε-binding domain places TANK among a set of adaptors (with SINTBAD and NAP1) that couple these kinases to virus-activated, IRF-dependent transcription [PMID:10581243, PMID:17568778]. As part of a TRAF3-containing complex it assembles TBK1/IKKε to drive IRF3/7 phosphorylation in selected TLR pathways [PMID:17823124]; genetic studies refine this role, showing TANK is essential for IKKε activation and for cross-talk between IKK-related and canonical IKKs while being dispensable for IRF3 phosphorylation and IFNβ production in macrophages [PMID:21949249]. TANK also functions as a brake on NF-κB: it recruits PLK1 to a TANK·NEMO complex to suppress NEMO ubiquitination and TNF-induced IKK activation [PMID:20484576], assembles a TANK·MCPIP1·USP10 complex that deubiquitinates TRAF6 to resolve genotoxic, IL-1β, and LPS-driven NF-κB signaling [PMID:25861989], and limits non-canonical NF-κB downstream of TACI [PMID:20394400]. Its activity is dynamically tuned by post-translational modification—IKKβ, IKKε, and TBK1 phosphorylation, K63-linked and mono-ubiquitination, and SUMOylation at the conserved Lys282, the last weakening the TANK–IKKε interaction to relieve TANK's inhibitory function during TLR7 signaling [PMID:10759890, PMID:16336209, PMID:17823124, PMID:21212807, PMID:22330071]. Knockout mice reveal physiological roles as a negative regulator of RANKL-induced osteoclastogenesis, where TANK loss elevates TRAF6 ubiquitination and causes trabecular bone loss [PMID:22773835]. TANK is also a target of viral immune evasion: SARS coronavirus M protein blocks TRAF3·TANK·TBK1/IKKε complex assembly, and multiple picornavirus 3C proteases cleave TANK to relieve its restraint on TRAF6-mediated NF-κB signaling [PMID:19380580, PMID:26363073, PMID:28566380].","teleology":[{"year":1996,"claim":"Established that TANK physically links to the TRAF machinery and can negatively regulate TRAF-driven NF-κB, defining it as an intrinsic modulator of TRAF function.","evidence":"Yeast two-hybrid, Co-IP, and NF-κB reporter assays identifying binding to TRAF-C domains of TRAF1/2/3 and inhibition of TRAF2-mediated NF-κB","pmids":["8710854","8608943"],"confidence":"High","gaps":["Whether TANK is activating or inhibitory in physiological context not resolved by overexpression","Endogenous stoichiometry with TRAFs unknown"]},{"year":1996,"claim":"Resolved the dual activating/inhibitory behavior by mapping it to TANK domains, showing an autoinhibitory C-terminus relieved by TRAF2 binding.","evidence":"Cotransfection NF-κB reporter assays with deletion mutants","pmids":["8608943"],"confidence":"High","gaps":["Molecular basis of the C-terminal autoinhibition not structurally defined","Native trigger for switching states unknown"]},{"year":1999,"claim":"Identified the TBK1 kinase as a TANK partner and showed a TBK1-TANK-TRAF2 ternary complex is required for kinase activity, placing TANK upstream of the IKK complex.","evidence":"Reciprocal Co-IP, kinase-dead dominant-negative constructs, NF-κB reporter epistasis","pmids":["10581243"],"confidence":"High","gaps":["How complex assembly activates TBK1 catalytically not defined","Receptor inputs that nucleate the complex not mapped"]},{"year":2000,"claim":"Demonstrated kinase-driven regulation of TANK assembly: IKKε phosphorylates TANK to release TRAF2, providing a feedback mechanism.","evidence":"Yeast two-hybrid, in vitro kinase assay, Co-IP, dominant-negative overexpression","pmids":["10759890"],"confidence":"Medium","gaps":["Phosphosite residues not precisely mapped","Single-lab in vitro phosphorylation, not validated in cells under physiological stimulation"]},{"year":2002,"claim":"Provided atomic-level basis for TANK-TRAF recognition, showing TANK uses a boomerang-shaped PxQxT motif competing with receptor tails for the TRAF crevice.","evidence":"X-ray crystallography of TANK peptide bound to TRAF3, mutagenesis, ITC, competition assays","pmids":["12005438"],"confidence":"High","gaps":["Structure limited to a peptide, not full-length TANK","Conformational dynamics of the intact complex unaddressed"]},{"year":2006,"claim":"Defined a C-terminal zinc finger mediating TNF-induced recruitment to the IKK complex and IKKβ phosphorylation that regulates NEMO binding, linking TANK to canonical TNF signaling.","evidence":"Co-IP, RNAi knockdown, NF-κB reporter assays, domain mapping","pmids":["16336209"],"confidence":"Medium","gaps":["IKKβ phosphosite not mapped","Single-lab RNAi, mechanism of p65 transactivation modulation unclear"]},{"year":2006,"claim":"Identified TFG as a TANK interactor enhancing NF-κB activation, expanding the TANK interactome.","evidence":"Yeast two-hybrid, Co-IP, NF-κB reporter assay","pmids":["16547966"],"confidence":"Low","gaps":["Single Co-IP/Y2H in one lab with limited TANK-specific follow-up","Physiological relevance of TFG-TANK axis untested"]},{"year":2007,"claim":"Established TANK as a scaffold assembling TBK1/IKKε within a TRAF3 complex for IRF3/7 phosphorylation, and characterized LPS-induced phosphorylation and K63-ubiquitination.","evidence":"Co-IP, in vitro kinase assay, RNAi, ubiquitin linkage analysis","pmids":["17823124"],"confidence":"High","gaps":["Ubiquitin ligase responsible for K63 chains not identified","Pathway selectivity (some but not all TLRs) not fully explained"]},{"year":2007,"claim":"Showed TANK is one of several redundant adaptors (with SINTBAD, NAP1) sharing a conserved TBK1/IKKi-binding domain, contextualizing its role in virus-activated signaling.","evidence":"Co-IP, TBD competition assays, siRNA, IRF-dependent reporter assays","pmids":["17568778"],"confidence":"High","gaps":["Functional non-redundancy among the three adaptors not resolved","Determinants of adaptor selection in vivo unknown"]},{"year":2009,"claim":"Revealed TANK as a target of viral immune evasion, with SARS-CoV M protein blocking TRAF3·TANK·TBK1/IKKε complex assembly to suppress interferon.","evidence":"Co-IP, IRF-dependent reporter assays, overexpression","pmids":["19380580"],"confidence":"Medium","gaps":["Direct binding partner of M protein within the complex not pinpointed","Overexpression-based, single lab"]},{"year":2010,"claim":"Defined a negative-regulatory mechanism whereby TANK scaffolds PLK1 into a complex with NEMO to suppress NEMO ubiquitination and TNF-induced IKK activation.","evidence":"Reciprocal Co-IP, ubiquitination assays, NF-κB reporter and DNA-binding assays","pmids":["20484576"],"confidence":"Medium","gaps":["Whether PLK1 modifies NEMO directly or via recruited factors unclear","Single lab"]},{"year":2010,"claim":"Extended TANK's inhibitory role to non-canonical NF-κB, showing it restrains NIK by promoting cIAP1-mediated NIK ubiquitination downstream of TACI.","evidence":"siRNA knockdown, Co-IP, ubiquitination assays, NF-κB reporter assays","pmids":["20394400"],"confidence":"Medium","gaps":["Mechanism by which TANK inhibits TRAF2-mediated cIAP1 inactivation not detailed","Single lab, siRNA-based"]},{"year":2011,"claim":"Genetic knockout dissected TANK's roles, showing it is essential for IKKε activation and IKK-related/canonical IKK cross-talk but dispensable for IRF3 phosphorylation and IFNβ production in macrophages.","evidence":"TANK knockout macrophages, kinase activity assays, Co-IP, IRF3 phosphorylation, IFNβ ELISA","pmids":["21949249"],"confidence":"High","gaps":["Redundancy with SINTBAD/NAP1 in IFN production not directly tested here","Cell-type specificity of the dispensability for IFN unclear"]},{"year":2011,"claim":"Identified SUMOylation at conserved Lys282, induced by IKKε/TBK1, as a switch that weakens TANK-IKKε binding to relieve TANK's negative function during TLR7 signaling.","evidence":"SUMOylation assays, K282 mutagenesis, Co-IP, reconstitution in TANK-deficient cells, reporter assays","pmids":["21212807","23201825"],"confidence":"High","gaps":["SUMO ligase responsible not identified","Interplay between SUMOylation and phosphorylation/ubiquitination not integrated"]},{"year":2012,"claim":"Knockout mice revealed TANK as a physiological negative regulator of osteoclastogenesis, restraining TRAF6 ubiquitination and protecting against bone loss.","evidence":"TANK knockout mice, osteoclast differentiation, ubiquitination and NF-κB assays, bone histomorphometry","pmids":["22773835"],"confidence":"High","gaps":["Molecular mechanism limiting TRAF6 ubiquitination not fully defined here","Relationship to the later USP10 deubiquitination mechanism not linked"]},{"year":2012,"claim":"Showed mono-ubiquitination of TANK (by Pellino3 downstream of oxLDL/SR-A1) blocks TBK1 recruitment to TRAF3, defining an ubiquitin-based control of TANK scaffold function.","evidence":"Co-IP, ubiquitination assays, RNAi, IRF3 activation assays","pmids":["22330071"],"confidence":"Medium","gaps":["Mono-ubiquitination acceptor site on TANK not mapped","Single lab"]},{"year":2015,"claim":"Defined a TANK·MCPIP1·USP10 complex that deubiquitinates TRAF6 to resolve genotoxic, IL-1β, and LPS-driven NF-κB, giving a molecular mechanism for TANK's negative regulation.","evidence":"CRISPR/Cas9 knockout, Co-IP, ubiquitination assays, NF-κB reporter assays","pmids":["25861989"],"confidence":"High","gaps":["How DNA damage signals reach the cytoplasmic complex not detailed","Stoichiometry and assembly order of the trimeric complex unknown"]},{"year":2017,"claim":"Provided structural and quantitative detail of TANK-TRAF1 binding, confirming the Px(Q/E)xT motif and comparable micromolar affinity for TRAF1 and TRAF2.","evidence":"X-ray crystallography (PDB 5H10), ITC, quantitative binding assays","pmids":["28155233"],"confidence":"High","gaps":["Functional consequence of TANK-TRAF1 binding specifically not addressed","Peptide-level structure only"]},{"year":2017,"claim":"Established TANK as a recurrent picornavirus protease target, with EMCV and Seneca Valley virus 3C proteases cleaving TANK to relieve its restraint on TRAF6-mediated NF-κB and impair IFN regulation.","evidence":"In vitro protease cleavage assays, cleavage-site mutagenesis, NF-κB and IFN reporter assays","pmids":["26363073","28566380"],"confidence":"Medium","gaps":["Cleavage demonstrated largely in vitro/overexpression","Consequences for viral fitness in vivo not established"]},{"year":2019,"claim":"Extended TANK's physiological scope to the nervous system, linking it to cortical NF-κB-dependent neuroimmune signaling and alcohol/anxiety behaviors.","evidence":"TANK knockout mice, behavioral assays, NF-κB activation in brain tissue","pmids":["30721969"],"confidence":"Medium","gaps":["Neuronal cell-type-specific contribution not dissected","Causal link between cortical NF-κB and behavior correlative"]},{"year":2022,"claim":"Defined a PLK1/TANK/NF-κB axis in intestinal epithelium, where TANK opposes PLK1's protective effect against LPS-induced epithelial injury, mitochondrial dysfunction, and apoptosis.","evidence":"Co-IP, siRNA knockdown, NF-κB and mitochondrial/apoptosis assays, in vivo CLP model","pmids":["36581806"],"confidence":"Medium","gaps":["Mechanistic relationship to the earlier PLK1-NEMO model not integrated","Single lab"]},{"year":null,"claim":"How TANK's opposing activating and inhibitory functions are selected within a given pathway, cell type, and post-translational modification state remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No integrated model linking phosphorylation, ubiquitination, and SUMOylation to functional switching","Tissue-specific roles (bone, brain, gut) lack a unifying mechanism","No full-length TANK structure to explain conformational regulation"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,7,8,16]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,10,14,16]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[2,7,8]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,2]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[7,8,12,13]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2,10,16]}],"complexes":["TBK1·TANK·TRAF2 complex","TRAF3·TANK·TBK1/IKKε complex","TANK·MCPIP1·USP10 complex","PLK1·TANK·NEMO complex"],"partners":["TRAF2","TRAF3","TRAF1","TBK1","IKKΕ","NEMO","PLK1","USP10"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q92844","full_name":"TRAF family member-associated NF-kappa-B activator","aliases":["TRAF-interacting protein","I-TRAF"],"length_aa":425,"mass_kda":47.8,"function":"Adapter protein involved in I-kappa-B-kinase (IKK) regulation which constitutively binds TBK1 and IKBKE playing a role in antiviral innate immunity. Acts as a regulator of TRAF function by maintaining them in a latent state. Blocks TRAF2 binding to LMP1 and inhibits LMP1-mediated NF-kappa-B activation. Negatively regulates NF-kappaB signaling and cell survival upon DNA damage (PubMed:25861989). Plays a role as an adapter to assemble ZC3H12A, USP10 in a deubiquitination complex which plays a negative feedback response to attenuate NF-kappaB activation through the deubiquitination of IKBKG or TRAF6 in response to interleukin-1-beta (IL1B) stimulation or upon DNA damage (PubMed:25861989). Promotes UBP10-induced deubiquitination of TRAF6 in response to DNA damage (PubMed:25861989). 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Study of the Bulk Tank Milk Microbiota Reveals Major Temporal Shifts in Composition.","date":"2021","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/33708181","citation_count":16,"is_preprint":false},{"pmid":"23139637","id":"PMC_23139637","title":"TRAF family member-associated NF-κB activator (TANK) induced by RANKL negatively regulates osteoclasts survival and function.","date":"2012","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/23139637","citation_count":15,"is_preprint":false},{"pmid":"22330071","id":"PMC_22330071","title":"OxLDL inhibits LPS-induced IFNβ expression by Pellino3- and IRAK1/4-dependent modification of TANK.","date":"2012","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/22330071","citation_count":15,"is_preprint":false},{"pmid":"23201825","id":"PMC_23201825","title":"TRAF family member-associated NF-kappa B activator (TANK) expression increases in injured sensory neurons and is transcriptionally regulated by Sox11.","date":"2012","source":"Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/23201825","citation_count":15,"is_preprint":false},{"pmid":"28155233","id":"PMC_28155233","title":"Molecular basis for TANK recognition by TRAF1 revealed by the crystal structure of TRAF1/TANK complex.","date":"2017","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/28155233","citation_count":15,"is_preprint":false},{"pmid":"34660604","id":"PMC_34660604","title":"Dual Regulation of Tank Binding Kinase 1 by BRG1 in Hepatocytes Contributes to Reactive Oxygen Species Production.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34660604","citation_count":15,"is_preprint":false},{"pmid":"39030571","id":"PMC_39030571","title":"Targeting TANK-binding kinase 1 attenuates painful diabetic neuropathy via inhibiting microglia pyroptosis.","date":"2024","source":"Cell communication and signaling : 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American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/38329325","citation_count":13,"is_preprint":false},{"pmid":"31560090","id":"PMC_31560090","title":"The transient expression of CHIKV VLP in large stirred tank bioreactors.","date":"2019","source":"Cytotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/31560090","citation_count":13,"is_preprint":false},{"pmid":"22230979","id":"PMC_22230979","title":"Somatic cells count in cow's bulk tank milk.","date":"2012","source":"The Journal of veterinary medical science","url":"https://pubmed.ncbi.nlm.nih.gov/22230979","citation_count":13,"is_preprint":false},{"pmid":"34248923","id":"PMC_34248923","title":"Human Metapneumovirus Induces IRF1 via TANK-Binding Kinase 1 and Type I IFN.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34248923","citation_count":12,"is_preprint":false},{"pmid":"30818760","id":"PMC_30818760","title":"Cell Metabolism Control Through O-GlcNAcylation of STAT5: A Full or Empty Fuel Tank Makes a Big Difference for Cancer Cell Growth and Survival.","date":"2019","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30818760","citation_count":12,"is_preprint":false},{"pmid":"29653159","id":"PMC_29653159","title":"Phosphorylation-induced changes in the energetic frustration in human Tank binding kinase 1.","date":"2018","source":"Journal of theoretical biology","url":"https://pubmed.ncbi.nlm.nih.gov/29653159","citation_count":12,"is_preprint":false},{"pmid":"20184394","id":"PMC_20184394","title":"TACI induces cIAP1-mediated ubiquitination of NIK by TRAF2 and TANK to limit non-canonical NF-kappaB signaling.","date":"2010","source":"Journal of receptor and signal transduction research","url":"https://pubmed.ncbi.nlm.nih.gov/20184394","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":49793,"output_tokens":6150,"usd":0.120815,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14951,"output_tokens":6171,"usd":0.114515,"stage2_stop_reason":"end_turn"},"total_usd":0.23533,"stage1_batch_id":"msgbatch_017qqVz33Ga63SCDwzb35XPb","stage2_batch_id":"msgbatch_01Scr2Ei9x4HJMiRGxwrmvLK","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1996,\n      \"finding\": \"TANK (I-TRAF) was identified as a novel intracellular protein that binds to the conserved TRAF-C domain of TRAF1, TRAF2, and TRAF3. Overexpression of I-TRAF inhibits TRAF2-mediated NF-κB activation downstream of CD40 and both TNF receptors, suggesting TANK acts as a natural regulator of TRAF function by maintaining TRAFs in a latent state.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation, cotransfection NF-κB reporter assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — independently identified in two labs (Rothe et al. PMID:8710854 and Cheng & Baltimore PMID:8608943) using multiple orthogonal methods including yeast two-hybrid, Co-IP, and functional reporter assays\",\n      \"pmids\": [\"8710854\", \"8608943\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"TANK and TRAF2 synergistically activate NF-κB in cotransfection experiments. This synergistic activation requires both the amino-terminal portion of TANK and the RING finger domain of TRAF2. TANK has an internally inhibitory carboxyl terminus, and TRAF2 binding overcomes this internal inhibition.\",\n      \"method\": \"Cotransfection NF-κB luciferase reporter assays, deletion mutagenesis\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional reporter assays combined with domain mutagenesis in a focused mechanistic study, replicated by related findings in PMID:8710854\",\n      \"pmids\": [\"8608943\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"TANK interacts with TBK1 (TANK-binding kinase 1), a novel IKK-related kinase. TBK1, TANK, and TRAF2 form a ternary complex, and complex formation is required for TBK1 kinase activity. Kinase-inactive TBK1 inhibits TANK-mediated NF-κB activation but does not block TNF-α, IL-1, or CD40 signaling, positioning the TBK1-TANK-TRAF2 complex upstream of NIK and the IKK complex.\",\n      \"method\": \"Co-immunoprecipitation, kinase-dead dominant-negative constructs, NF-κB reporter assays, genetic epistasis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP identifying the ternary complex, dominant-negative kinase assays, and epistasis experiments all in one study; foundational result replicated in subsequent work\",\n      \"pmids\": [\"10581243\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"IKK-i (IKKε) phosphorylates TANK/I-TRAF in vitro in the middle portion of TANK that associates with TRAF2. Following IKK-i-mediated phosphorylation of TANK, TRAF2 is released from the TANK/TRAF2 complex. The N-terminal domain of TANK mediates its interaction with the C-terminal portion of IKK-i, and NF-κB activation by IKK-i is blocked by co-expression of the N-terminal domain of TANK.\",\n      \"method\": \"Yeast two-hybrid (IKK-i/TANK interaction), in vitro kinase assay, co-immunoprecipitation, dominant-negative overexpression\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro kinase assay with domain mapping plus Co-IP and functional assays, single lab\",\n      \"pmids\": [\"10759890\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Crystal structure of a TANK peptide bound to TRAF3 was determined, showing that TANK and CD40 bind to the same crevice on TRAF3. TANK presents a boomerang-like structure at the recognition motif PxQxT that is distinct from the hairpin loop of CD40. Mutagenesis confirmed critical TANK contact residues for binding to TRAF3 and TRAF2; TANK competes with CD40 for the TRAF binding site as demonstrated by isothermal titration calorimetry and competition assays.\",\n      \"method\": \"X-ray crystallography, site-directed mutagenesis, isothermal titration calorimetry, competition binding assays\",\n      \"journal\": \"Structure (London, England : 1993)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus mutagenesis plus quantitative binding assays in one study\",\n      \"pmids\": [\"12005438\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"TNF-α stimulation promotes TANK recruitment to the IKK complex via a newly characterized C-terminal zinc finger in TANK. IKKβ phosphorylates TANK upon TNF-α stimulation, and this phosphorylation negatively regulates TANK binding to NEMO. Reduced TANK expression (by RNAi) attenuates TNF-α-mediated induction of a subset of NF-κB target genes through decreased p65 transactivation potential.\",\n      \"method\": \"Co-immunoprecipitation, RNAi knockdown, NF-κB reporter assays, domain mapping\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for complex mapping plus functional RNAi knockdown, single lab with multiple methods\",\n      \"pmids\": [\"16336209\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"TFG (TRK-fused gene) was identified as a novel TANK-interacting protein through yeast two-hybrid screening and confirmed by in vitro and in vivo co-immunoprecipitation. TFG and NEMO may be part of the same high molecular weight complex. TFG enhances TANK-induced NF-κB activation.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, NF-κB reporter assay\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP/yeast two-hybrid in one lab with limited mechanistic follow-up specific to TANK\",\n      \"pmids\": [\"16547966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"TANK was identified as a scaffold protein that assembles TBK1/IKKε complexes required for IRF3/7 phosphorylation in some (but not all) Toll-like receptor-dependent signaling pathways. TANK is part of a TRAF3-containing complex. Upon LPS stimulation, TANK is heavily phosphorylated by TBK1-IKKε and undergoes Lys63-linked polyubiquitination in a manner that does not require TBK1-IKKε kinase activity.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay, RNAi knockdown, ubiquitin linkage analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, kinase assay, RNAi, ubiquitin linkage) in one focused study of TANK's scaffold function\",\n      \"pmids\": [\"17823124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"SINTBAD, NAP1, and TANK share a conserved TBK1/IKKi-binding domain (TBD) predicted to form an alpha-helix with residues essential for kinase binding on one face. Isolated TBDs from each adaptor compete with each other for binding to TBK1 and prevent poly(I:C)-induced IRF-dependent transcription, establishing TANK as one of multiple adaptors linking TBK1/IKKi to virus-activated signaling.\",\n      \"method\": \"Co-immunoprecipitation, competition assays, siRNA knockdown, IRF-dependent transcription reporter assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — TBD competition assays plus RNAi plus reporter assays; TANK's TBK1-binding domain structurally defined and functionally validated\",\n      \"pmids\": [\"17568778\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"SARS coronavirus M protein physically associates with TRAF3, TBK1, IKKε, and RIG-I, and prevents formation of the TRAF3·TANK·TBK1/IKKε complex, thereby inhibiting TBK1/IKKε-dependent activation of IRF3/IRF7 and type I interferon production. M protein has no influence when IRF3 or IRF7 are overexpressed directly, indicating the block is upstream at complex assembly.\",\n      \"method\": \"Co-immunoprecipitation, IRF-dependent transcription reporter assay, overexpression studies\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating complex disruption plus functional reporter assays, single lab\",\n      \"pmids\": [\"19380580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"TANK directly interacts with PLK1 (polo-like kinase 1). PLK1, TANK, and NEMO (IKKγ) form a ternary complex in vivo. PLK1 negatively regulates TNF-induced IKK activation by inhibiting ubiquitination of NEMO, establishing TANK as a scaffold that recruits PLK1 to suppress NF-κB signaling.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, NF-κB reporter assays, NF-κB DNA-binding assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP for ternary complex plus functional ubiquitination and reporter assays, single lab\",\n      \"pmids\": [\"20484576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"TANK participates in limiting non-canonical NF-κB signaling downstream of TACI. TANK deficiency (by siRNA) impairs TACI-dependent inhibition of NF-κB2 p100 processing. TANK inhibits TRAF2-mediated cIAP1 inactivation and thereby facilitates cIAP1-mediated ubiquitination of NIK to suppress non-canonical NF-κB signaling.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation, ubiquitination assays, NF-κB reporter assays\",\n      \"journal\": \"Journal of receptor and signal transduction research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA loss-of-function with defined pathway phenotype plus Co-IP, single lab\",\n      \"pmids\": [\"20394400\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"TANK is required for the activation of IKKε and for optimal activation of TBK1 in macrophages stimulated through both MyD88- and TRIF-dependent TLR pathways. TANK is required for the interaction between IKK-related kinases and canonical IKKs, enabling IKK-related kinases to negatively regulate canonical IKKs. In TANK−/− macrophages, IKKε activation is abolished, TBK1 activation is reduced, but IRF3 phosphorylation and IFNβ production remain normal.\",\n      \"method\": \"TANK knockout macrophages, kinase activity assays, co-immunoprecipitation, IRF3 phosphorylation assays, IFNβ ELISA\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic KO with multiple phenotypic readouts and Co-IP, separating TANK's roles in IKK cross-talk vs. IFN production\",\n      \"pmids\": [\"21949249\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"SUMOylation of TANK at the evolutionarily conserved Lys282 is triggered by IKKε and TBK1 kinase activities. Stimulation of TLR7 induces TANK SUMOylation, which weakens the interaction between TANK and IKKε, thereby relieving TANK's negative function on TLR7 signal propagation. Reconstitution experiments show that absence of TANK SUMOylation impairs inducible expression of TLR7-dependent target genes.\",\n      \"method\": \"SUMOylation assays, site-directed mutagenesis (K282), co-immunoprecipitation, reconstitution in TANK-deficient cells, reporter assays\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis of the SUMO site plus Co-IP plus reconstitution experiments, multiple methods in one study\",\n      \"pmids\": [\"21212807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TANK is a negative regulator of osteoclast differentiation. TANK expression is upregulated during RANKL-induced osteoclastogenesis. Tank−/− cells show increased osteoclastogenesis with increased TRAF6 ubiquitination and enhanced canonical NF-κB activation in response to RANKL. Tank−/− mice develop severe trabecular bone loss due to enhanced bone erosion.\",\n      \"method\": \"TANK knockout mouse model, osteoclast differentiation assays, ubiquitination assays, NF-κB activation assays, bone histomorphometry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO with defined cellular phenotype, mechanistic ubiquitination assays, and in vivo bone phenotype\",\n      \"pmids\": [\"22773835\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"In the context of virus-triggered signaling, TANK mono-ubiquitination (mediated by Pellino3 downstream of IRAK1/4 via SR-A1) inhibits TBK1 recruitment to TRAF3, thereby blocking TBK1-TRAF3 interaction and subsequent IRF3 activation and IFNβ expression. oxLDL stimulates this TANK mono-ubiquitination pathway.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, RNAi knockdown, IRF3 activation assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP showing TBK1-TRAF3 disruption plus ubiquitination and RNAi assays, single lab\",\n      \"pmids\": [\"22330071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TANK negatively regulates genotoxic NF-κB activation by facilitating USP10-dependent deubiquitination of TRAF6. TANK forms a complex with MCPIP1 (ZC3H12A) and the deubiquitinase USP10, which is essential for TRAF6 deubiquitination and resolution of DNA damage-induced NF-κB activation. CRISPR/Cas9 deletion of TANK in human cells enhances NF-κB activation after genotoxic treatment. The TANK-MCPIP1-USP10 complex also decreases TRAF6 ubiquitination downstream of IL-1β and LPS.\",\n      \"method\": \"CRISPR/Cas9 knockout, co-immunoprecipitation, ubiquitination assays, NF-κB reporter assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CRISPR KO plus Co-IP identifying trimeric complex plus functional ubiquitination and NF-κB assays in multiple stimulation contexts\",\n      \"pmids\": [\"25861989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Encephalomyocarditis virus (EMCV) 3C protease cleaves TANK at Q197 and Q291, dependent on 3C cysteine protease activity. Cleavage of TANK by EMCV 3C impairs TANK's ability to inhibit TRAF6-mediated NF-κB signaling. Several other picornavirus proteases (FMDV, PRRSV, EAV) also cleave TANK.\",\n      \"method\": \"In vitro protease cleavage assays, site-directed mutagenesis, NF-κB reporter assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro cleavage assay with mutagenesis of cleavage sites plus functional NF-κB assay, single lab\",\n      \"pmids\": [\"26363073\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Crystal structure of the TRAF1/TANK peptide complex was determined (PDB: 5H10). TANK binds TRAF1 using the minor minimal consensus motif Px(Q/E)xT. Quantitative interaction experiments showed TANK peptide interacts with TRAF1 and TRAF2 with similar micromolar affinity.\",\n      \"method\": \"X-ray crystallography, isothermal titration calorimetry, quantitative binding assays\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus quantitative binding measurements, single lab but Tier 1 method\",\n      \"pmids\": [\"28155233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Seneca Valley virus 3C protease mediates cleavage of TANK (along with MAVS and TRIF) at specific sites requiring 3C protease activity. Cleaved TANK loses the ability to regulate PRR-mediated IFN production, and TANK cleavage facilitates TRAF6-induced NF-κB activation.\",\n      \"method\": \"In vitro protease cleavage assays, IFN production assays, NF-κB reporter assays\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro cleavage assay with functional IFN and NF-κB readouts, single lab\",\n      \"pmids\": [\"28566380\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"In the context of TLR7 signaling, TANK SUMOylation (induced by TBK1/IKKε) weakens the TANK-IKKε interaction, thereby relieving TANK's constitutive negative function on downstream signal propagation. Separately, TANK expression is up-regulated in injured sensory neurons (DRG) and is transcriptionally regulated by the injury-associated transcription factor Sox11 via direct binding to two Sox motifs in the TANK 5'-UTR.\",\n      \"method\": \"Chromatin immunoprecipitation, luciferase reporter with Sox site mutagenesis, Sox11 siRNA knockdown, quantitative PCR\",\n      \"journal\": \"Neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus mutagenesis of promoter sites plus siRNA knockdown, single lab; establishes Sox11 as a transcriptional regulator of TANK\",\n      \"pmids\": [\"23201825\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TANK knockout mice display altered alcohol drinking behavior and enhanced anxiety-related behavior, and show increased NF-κB activation in the insular cortex following alcohol exposure, suggesting TANK regulates cortical NF-κB-dependent neuroimmune signaling in the context of aversive emotional processing.\",\n      \"method\": \"Tank knockout mouse model, behavioral assays, NF-κB activation assays in brain tissue\",\n      \"journal\": \"Cerebral cortex (New York, N.Y. : 1991)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with behavioral phenotype and biochemical NF-κB readout in brain tissue, single lab\",\n      \"pmids\": [\"30721969\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PLK1 co-localizes and physically interacts with TANK in intestinal epithelial (Caco-2) cells. TANK overexpression impairs the protective effect of PLK1 on LPS-induced intestinal epithelial injury and NF-κB activation, and TANK siRNA knockdown suppresses NF-κB signaling and ameliorates mitochondrial dysfunction and apoptosis induced by LPS. This defines a PLK1/TANK/NF-κB axis regulating intestinal barrier function.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, NF-κB signaling assays, mitochondrial dynamics assays, apoptosis assays, in vivo CLP model\",\n      \"journal\": \"Molecular medicine (Cambridge, Mass.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus siRNA plus in vivo model, single lab with multiple methods\",\n      \"pmids\": [\"36581806\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TANK (I-TRAF) is a scaffold/adaptor protein that binds to the TRAF-C domains of TRAF1, TRAF2, and TRAF3 via a PxQxT-like motif, forms a ternary complex with TBK1 and TRAF2 that is required for TBK1 kinase activity and NF-κB activation, and assembles the TRAF3·TANK·TBK1/IKKε complex essential for IRF3/7 phosphorylation and type I interferon induction; TANK also acts as a negative regulator of NF-κB by recruiting PLK1 to inhibit NEMO ubiquitination and by facilitating USP10-dependent deubiquitination of TRAF6, and its activity is fine-tuned by IKKε/TBK1-induced phosphorylation and SUMOylation at K282, the latter relieving its inhibitory function during TLR7 signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TANK (I-TRAF) is an intracellular scaffold/adaptor protein that integrates TRAF-family signaling to both activate and restrain NF-κB and type I interferon induction [#0, #2]. It engages the conserved TRAF-C domains of TRAF1, TRAF2, and TRAF3 through a boomerang-shaped PxQxT-like recognition motif that occupies the same TRAF crevice used by receptor tails such as CD40, allowing TANK to compete for TRAF binding [#0, #4, #18]. Through an N-terminal region TANK nucleates a ternary complex with the IKK-related kinase TBK1 and TRAF2 that is required for TBK1 kinase activity, and a shared TBK1/IKKε-binding domain places TANK among a set of adaptors (with SINTBAD and NAP1) that couple these kinases to virus-activated, IRF-dependent transcription [#2, #8]. As part of a TRAF3-containing complex it assembles TBK1/IKKε to drive IRF3/7 phosphorylation in selected TLR pathways [#7]; genetic studies refine this role, showing TANK is essential for IKKε activation and for cross-talk between IKK-related and canonical IKKs while being dispensable for IRF3 phosphorylation and IFNβ production in macrophages [#12]. TANK also functions as a brake on NF-κB: it recruits PLK1 to a TANK·NEMO complex to suppress NEMO ubiquitination and TNF-induced IKK activation [#10], assembles a TANK·MCPIP1·USP10 complex that deubiquitinates TRAF6 to resolve genotoxic, IL-1β, and LPS-driven NF-κB signaling [#16], and limits non-canonical NF-κB downstream of TACI [#11]. Its activity is dynamically tuned by post-translational modification—IKKβ, IKKε, and TBK1 phosphorylation, K63-linked and mono-ubiquitination, and SUMOylation at the conserved Lys282, the last weakening the TANK–IKKε interaction to relieve TANK's inhibitory function during TLR7 signaling [#3, #5, #7, #13, #15]. Knockout mice reveal physiological roles as a negative regulator of RANKL-induced osteoclastogenesis, where TANK loss elevates TRAF6 ubiquitination and causes trabecular bone loss [#14]. TANK is also a target of viral immune evasion: SARS coronavirus M protein blocks TRAF3·TANK·TBK1/IKKε complex assembly, and multiple picornavirus 3C proteases cleave TANK to relieve its restraint on TRAF6-mediated NF-κB signaling [#9, #17, #19].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Established that TANK physically links to the TRAF machinery and can negatively regulate TRAF-driven NF-κB, defining it as an intrinsic modulator of TRAF function.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, and NF-κB reporter assays identifying binding to TRAF-C domains of TRAF1/2/3 and inhibition of TRAF2-mediated NF-κB\",\n      \"pmids\": [\"8710854\", \"8608943\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether TANK is activating or inhibitory in physiological context not resolved by overexpression\", \"Endogenous stoichiometry with TRAFs unknown\"]\n    },\n    {\n      \"year\": 1996,\n      \"claim\": \"Resolved the dual activating/inhibitory behavior by mapping it to TANK domains, showing an autoinhibitory C-terminus relieved by TRAF2 binding.\",\n      \"evidence\": \"Cotransfection NF-κB reporter assays with deletion mutants\",\n      \"pmids\": [\"8608943\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of the C-terminal autoinhibition not structurally defined\", \"Native trigger for switching states unknown\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Identified the TBK1 kinase as a TANK partner and showed a TBK1-TANK-TRAF2 ternary complex is required for kinase activity, placing TANK upstream of the IKK complex.\",\n      \"evidence\": \"Reciprocal Co-IP, kinase-dead dominant-negative constructs, NF-κB reporter epistasis\",\n      \"pmids\": [\"10581243\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How complex assembly activates TBK1 catalytically not defined\", \"Receptor inputs that nucleate the complex not mapped\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Demonstrated kinase-driven regulation of TANK assembly: IKKε phosphorylates TANK to release TRAF2, providing a feedback mechanism.\",\n      \"evidence\": \"Yeast two-hybrid, in vitro kinase assay, Co-IP, dominant-negative overexpression\",\n      \"pmids\": [\"10759890\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Phosphosite residues not precisely mapped\", \"Single-lab in vitro phosphorylation, not validated in cells under physiological stimulation\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Provided atomic-level basis for TANK-TRAF recognition, showing TANK uses a boomerang-shaped PxQxT motif competing with receptor tails for the TRAF crevice.\",\n      \"evidence\": \"X-ray crystallography of TANK peptide bound to TRAF3, mutagenesis, ITC, competition assays\",\n      \"pmids\": [\"12005438\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure limited to a peptide, not full-length TANK\", \"Conformational dynamics of the intact complex unaddressed\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined a C-terminal zinc finger mediating TNF-induced recruitment to the IKK complex and IKKβ phosphorylation that regulates NEMO binding, linking TANK to canonical TNF signaling.\",\n      \"evidence\": \"Co-IP, RNAi knockdown, NF-κB reporter assays, domain mapping\",\n      \"pmids\": [\"16336209\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"IKKβ phosphosite not mapped\", \"Single-lab RNAi, mechanism of p65 transactivation modulation unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identified TFG as a TANK interactor enhancing NF-κB activation, expanding the TANK interactome.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, NF-κB reporter assay\",\n      \"pmids\": [\"16547966\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP/Y2H in one lab with limited TANK-specific follow-up\", \"Physiological relevance of TFG-TANK axis untested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Established TANK as a scaffold assembling TBK1/IKKε within a TRAF3 complex for IRF3/7 phosphorylation, and characterized LPS-induced phosphorylation and K63-ubiquitination.\",\n      \"evidence\": \"Co-IP, in vitro kinase assay, RNAi, ubiquitin linkage analysis\",\n      \"pmids\": [\"17823124\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitin ligase responsible for K63 chains not identified\", \"Pathway selectivity (some but not all TLRs) not fully explained\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed TANK is one of several redundant adaptors (with SINTBAD, NAP1) sharing a conserved TBK1/IKKi-binding domain, contextualizing its role in virus-activated signaling.\",\n      \"evidence\": \"Co-IP, TBD competition assays, siRNA, IRF-dependent reporter assays\",\n      \"pmids\": [\"17568778\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional non-redundancy among the three adaptors not resolved\", \"Determinants of adaptor selection in vivo unknown\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Revealed TANK as a target of viral immune evasion, with SARS-CoV M protein blocking TRAF3·TANK·TBK1/IKKε complex assembly to suppress interferon.\",\n      \"evidence\": \"Co-IP, IRF-dependent reporter assays, overexpression\",\n      \"pmids\": [\"19380580\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding partner of M protein within the complex not pinpointed\", \"Overexpression-based, single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined a negative-regulatory mechanism whereby TANK scaffolds PLK1 into a complex with NEMO to suppress NEMO ubiquitination and TNF-induced IKK activation.\",\n      \"evidence\": \"Reciprocal Co-IP, ubiquitination assays, NF-κB reporter and DNA-binding assays\",\n      \"pmids\": [\"20484576\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether PLK1 modifies NEMO directly or via recruited factors unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Extended TANK's inhibitory role to non-canonical NF-κB, showing it restrains NIK by promoting cIAP1-mediated NIK ubiquitination downstream of TACI.\",\n      \"evidence\": \"siRNA knockdown, Co-IP, ubiquitination assays, NF-κB reporter assays\",\n      \"pmids\": [\"20394400\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which TANK inhibits TRAF2-mediated cIAP1 inactivation not detailed\", \"Single lab, siRNA-based\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Genetic knockout dissected TANK's roles, showing it is essential for IKKε activation and IKK-related/canonical IKK cross-talk but dispensable for IRF3 phosphorylation and IFNβ production in macrophages.\",\n      \"evidence\": \"TANK knockout macrophages, kinase activity assays, Co-IP, IRF3 phosphorylation, IFNβ ELISA\",\n      \"pmids\": [\"21949249\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Redundancy with SINTBAD/NAP1 in IFN production not directly tested here\", \"Cell-type specificity of the dispensability for IFN unclear\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identified SUMOylation at conserved Lys282, induced by IKKε/TBK1, as a switch that weakens TANK-IKKε binding to relieve TANK's negative function during TLR7 signaling.\",\n      \"evidence\": \"SUMOylation assays, K282 mutagenesis, Co-IP, reconstitution in TANK-deficient cells, reporter assays\",\n      \"pmids\": [\"21212807\", \"23201825\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"SUMO ligase responsible not identified\", \"Interplay between SUMOylation and phosphorylation/ubiquitination not integrated\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Knockout mice revealed TANK as a physiological negative regulator of osteoclastogenesis, restraining TRAF6 ubiquitination and protecting against bone loss.\",\n      \"evidence\": \"TANK knockout mice, osteoclast differentiation, ubiquitination and NF-κB assays, bone histomorphometry\",\n      \"pmids\": [\"22773835\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism limiting TRAF6 ubiquitination not fully defined here\", \"Relationship to the later USP10 deubiquitination mechanism not linked\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed mono-ubiquitination of TANK (by Pellino3 downstream of oxLDL/SR-A1) blocks TBK1 recruitment to TRAF3, defining an ubiquitin-based control of TANK scaffold function.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, RNAi, IRF3 activation assays\",\n      \"pmids\": [\"22330071\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mono-ubiquitination acceptor site on TANK not mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined a TANK·MCPIP1·USP10 complex that deubiquitinates TRAF6 to resolve genotoxic, IL-1β, and LPS-driven NF-κB, giving a molecular mechanism for TANK's negative regulation.\",\n      \"evidence\": \"CRISPR/Cas9 knockout, Co-IP, ubiquitination assays, NF-κB reporter assays\",\n      \"pmids\": [\"25861989\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How DNA damage signals reach the cytoplasmic complex not detailed\", \"Stoichiometry and assembly order of the trimeric complex unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Provided structural and quantitative detail of TANK-TRAF1 binding, confirming the Px(Q/E)xT motif and comparable micromolar affinity for TRAF1 and TRAF2.\",\n      \"evidence\": \"X-ray crystallography (PDB 5H10), ITC, quantitative binding assays\",\n      \"pmids\": [\"28155233\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of TANK-TRAF1 binding specifically not addressed\", \"Peptide-level structure only\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Established TANK as a recurrent picornavirus protease target, with EMCV and Seneca Valley virus 3C proteases cleaving TANK to relieve its restraint on TRAF6-mediated NF-κB and impair IFN regulation.\",\n      \"evidence\": \"In vitro protease cleavage assays, cleavage-site mutagenesis, NF-κB and IFN reporter assays\",\n      \"pmids\": [\"26363073\", \"28566380\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Cleavage demonstrated largely in vitro/overexpression\", \"Consequences for viral fitness in vivo not established\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended TANK's physiological scope to the nervous system, linking it to cortical NF-κB-dependent neuroimmune signaling and alcohol/anxiety behaviors.\",\n      \"evidence\": \"TANK knockout mice, behavioral assays, NF-κB activation in brain tissue\",\n      \"pmids\": [\"30721969\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Neuronal cell-type-specific contribution not dissected\", \"Causal link between cortical NF-κB and behavior correlative\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a PLK1/TANK/NF-κB axis in intestinal epithelium, where TANK opposes PLK1's protective effect against LPS-induced epithelial injury, mitochondrial dysfunction, and apoptosis.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, NF-κB and mitochondrial/apoptosis assays, in vivo CLP model\",\n      \"pmids\": [\"36581806\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic relationship to the earlier PLK1-NEMO model not integrated\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How TANK's opposing activating and inhibitory functions are selected within a given pathway, cell type, and post-translational modification state remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No integrated model linking phosphorylation, ubiquitination, and SUMOylation to functional switching\", \"Tissue-specific roles (bone, brain, gut) lack a unifying mechanism\", \"No full-length TANK structure to explain conformational regulation\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 7, 8, 16]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 10, 14, 16]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [2, 7, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [7, 8, 12, 13]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2, 10, 16]}\n    ],\n    \"complexes\": [\n      \"TBK1·TANK·TRAF2 complex\",\n      \"TRAF3·TANK·TBK1/IKKε complex\",\n      \"TANK·MCPIP1·USP10 complex\",\n      \"PLK1·TANK·NEMO complex\"\n    ],\n    \"partners\": [\n      \"TRAF2\",\n      \"TRAF3\",\n      \"TRAF1\",\n      \"TBK1\",\n      \"IKKε\",\n      \"NEMO\",\n      \"PLK1\",\n      \"USP10\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":{"gene":"TANK","tier":"GROUNDING","verdict":"Evidence-grounding concern","subtype":"fabrication","uniprot_band":"medium","rules_fired":"R7","issue":"R7: fabricated (no corpus paper): 20394400"},"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}