Affinage

TIFA

TRAF-interacting protein with FHA domain-containing protein A · UniProt Q96CG3

Length
184 aa
Mass
21.4 kDa
Annotated
2026-06-10
46 papers in source corpus 33 papers cited in narrative 33 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

TIFA is an FHA-domain adaptor protein that serves as a central signaling hub coupling innate immune sensing of bacterial metabolites to NF-κB activation (PMID:26068852, PMID:28877472). In its resting state TIFA exists as an intrinsic dimer; phosphorylation at Thr9 generates a pThr9 motif that engages the FHA domain of a neighboring dimer, driving higher-order oligomerization that is the trigger for downstream signaling (PMID:22566686, PMID:26389808). Cytosolic detection of bacterial ADP-heptose/HBP metabolites by the upstream kinase ALPK1 catalyzes this Thr9 phosphorylation during infection by H. pylori, Shigella, Salmonella, and other organisms, nucleating large TIFA oligomeric complexes (TIFAsomes) that behave as liquid-liquid phase-separated condensates (PMID:28877472, PMID:28222186, PMID:38357697). Within these assemblies TIFA recruits TRAF6 to drive K63-linked polyubiquitination and TAK1/IKK-dependent classical NF-κB signaling, and engages TRAF2 to displace cIAP1 and activate the alternative NF-κB pathway (PMID:15492226, PMID:34328245, PMID:30378729). TIFA additionally bridges IRAK-1 to TRAF6 in IL-1 receptor signaling (PMID:10920205, PMID:12566447), and translocates to damaged chromatin upon genotoxic stress where its pThr9/FHA interaction and TRAF2 cooperation stimulate NEMO ubiquitination (PMID:29581234). Signaling is restrained by TIFAB, which heterodimerizes with monomeric TIFA to form a pseudo-dimer lacking the phosphorylation site and TRAF6-binding motif, blocking productive oligomer assembly (PMID:38442163, PMID:15047173), and activated TIFA is cleared via proteasomal, lysosomal, and TRAF2/TRAF6-dependent autophagic degradation (PMID:36965415, PMID:40696502). Through this pathway TIFA controls intestinal barrier gene expression and inflammasome priming, and its dysregulation promotes leukemic and colorectal cancer cell survival and proliferation (PMID:36036242, PMID:27965388, PMID:28069801, PMID:35635239).

Mechanistic history

Synthesis pass · year-by-year structured walk · 24 steps
  1. 2000 Medium

    Established TIFA as a TRAF6-associated adaptor selectively engaged downstream of IL-1 but not TNF, placing it in receptor-proximal innate immune signaling.

    Evidence Yeast two-hybrid and co-immunoprecipitation mapping the coiled-coil/TRAF6 ring-finger interaction

    PMID:10920205

    Open questions at the time
    • Did not define the activation trigger upstream of TIFA
    • IRAK absent from the complex left the IRAK-TRAF6 bridging unexplained
  2. 2002 Medium

    Showed TIFA also binds TRAF2 and is sufficient to activate NF-κB and AP-1 when overexpressed, broadening its connectivity to multiple TRAF effectors.

    Evidence Mammalian two-hybrid, Co-IP, and NF-κB/AP-1 reporter assays

    PMID:11798190

    Open questions at the time
    • Overexpression-driven activation did not establish the physiological stimulus
    • Distinct roles of TRAF2 vs TRAF6 binding not resolved
  3. 2003 High

    Defined TIFA as the bridge that recruits IRAK-1 to TRAF6 upon IL-1 stimulation, resolving how an upstream kinase links to the TRAF6 effector.

    Evidence Endogenous Co-IP, FHA/TRAF6-binding mutagenesis, and NF-κB/JNK reporters

    PMID:12566447

    Open questions at the time
    • The kinase phosphorylating TIFA itself was not identified
    • Mechanism converting binding into TRAF6 activation unknown
  4. 2004 High

    Demonstrated the biochemical mechanism by which TIFA activates IKK: oligomeric TIFA promotes K63-linked TRAF6 polyubiquitination and TAK1/IKK activation proteasome-independently.

    Evidence In vitro reconstitution with purified TIFA, TRAF6, TAK1, Ubc13-Uev1A and cell-free IKK assays

    PMID:15492226

    Open questions at the time
    • What drives TIFA into the active oligomeric form in cells was undefined
    • Structural basis of oligomerization unresolved
  5. 2004 Medium

    Identified TIFAB as a homolog that inhibits TIFA-mediated NF-κB activation, establishing negative regulation of the pathway.

    Evidence Co-IP and NF-κB reporter assays

    PMID:15047173

    Open questions at the time
    • Structural basis of inhibition inferred but not directly shown
    • Whether TIFAB acts on monomer vs oligomer not resolved at this stage
  6. 2007 High

    Connected TIFA to membrane trafficking via myosin VI binding and trans-Golgi localization, indicating roles beyond cytosolic NF-κB signaling.

    Evidence Yeast two-hybrid, in vitro binding, IF/EM localization, and RNAi phenotyping

    PMID:17635994

    Open questions at the time
    • Mechanistic link between trafficking role and NF-κB signaling unclear
    • How myosin VI binding relates to oligomerization not addressed
  7. 2012 High

    Pinpointed Thr9 phosphorylation and the pThr9-FHA inter-dimer interaction as the molecular switch driving TIFA oligomerization and NF-κB activation.

    Evidence Phosphorylation mapping, oligomerization assays, siRNA knockdown, and NF-κB reporters

    PMID:22566686

    Open questions at the time
    • The physiological kinase and stimulus were not identified
    • Higher-order complex architecture not visualized
  8. 2015 High

    Identified the bacterial metabolite HBP as the cytosolic ligand triggering TIFA-dependent NF-κB activation, defining the upstream stimulus.

    Evidence Genome-wide RNAi screen and cell-based NF-κB/oligomerization assays

    PMID:26068852

    Open questions at the time
    • The sensor/kinase coupling HBP to TIFA phosphorylation was not yet identified
  9. 2015 High

    Provided the structural mechanism for oligomerization, showing TIFA is an intrinsic dimer and pThr9-FHA contacts occur only between distinct dimers.

    Evidence X-ray crystallography of TIFA 1-150 and the pThr9 peptide complex with mutagenesis

    PMID:26389808

    Open questions at the time
    • Full higher-order oligomer/condensate structure not resolved
    • TRAF6 incorporation geometry not determined
  10. 2016 Medium

    Showed multiple host kinases (Aurora A, Akt) can phosphorylate TIFA Thr9 to drive NF-κB survival signaling and inflammasome priming, expanding TIFA into oncogenic and vascular contexts.

    Evidence Kinase assays, siRNA knockdown, Co-IP (TIFA-caspase-1), reporter assays, and xenograft/flow models

    PMID:27965388 PMID:28069801

    Open questions at the time
    • Relative contribution of each kinase in physiological settings unclear
    • Direct caspase-1 engagement mechanism not structurally defined
  11. 2017 High

    Identified ALPK1 as the ADP-heptose/HBP sensor kinase that phosphorylates TIFA Thr9 to nucleate TIFAsomes during enteric and gastric bacterial infection, completing the ligand-kinase-adaptor axis.

    Evidence CRISPR/Cas9 knockout, RNAi, mass spectrometry of TIFAsome components, IF, and bacterial mutants across H. pylori, Shigella, and Salmonella models

    PMID:28222186 PMID:28514661 PMID:28811347 PMID:28877472

    Open questions at the time
    • Quantitative biophysics of TIFAsome assembly not yet defined
    • How TIFA discriminates replication rate mechanistically partly inferred
  12. 2018 Medium

    Extended TIFA function to the DNA damage response, showing genotoxic stress drives TIFA pThr9-dependent chromatin accumulation and TRAF2-dependent NEMO ubiquitination.

    Evidence Subcellular fractionation, IF, Thr9 mutagenesis, ubiquitination assays, and siRNA knockdown

    PMID:29581234

    Open questions at the time
    • The kinase phosphorylating TIFA after DNA damage not identified
    • How TIFA targets damaged chromatin mechanistically unresolved
  13. 2018 High

    Defined the direct TIFA C-terminus/TRAF6 TRAF-domain interface at atomic resolution, mapping the salt-bridge contacts required for engagement.

    Evidence X-ray crystallography of TRAF6 with WT and mutant TIFA peptides plus Co-IP

    PMID:30378729

    Open questions at the time
    • Stoichiometry of TIFA-TRAF6 in the active condensate not defined
  14. 2020 High

    Confirmed conserved TIFA dimer architecture in mouse and modeled a higher-order TIFA-TRAF6 signaling complex, reinforcing the dimer-to-oligomer mechanism.

    Evidence Crystallography of mouse TIFA and phosphomimetic mutants, SEC, and SAXS

    PMID:32198460

    Open questions at the time
    • Solution oligomer beyond the dimer modeled but not directly visualized
  15. 2020 Medium

    Linked TIFA/NF-κB signaling to genome instability, showing the ALPK1/TIFA axis drives co-transcriptional R-loop formation and DNA double-strand breaks during H. pylori infection.

    Evidence CRISPR knockout, gastric organoids, R-loop/S9.6 detection, DNA fiber assays, and bacterial mutants

    PMID:33037203

    Open questions at the time
    • Direct molecular link from NF-κB output to R-loop induction not fully defined
    • Causal role of TIFA in carcinogenesis not established
  16. 2021 Medium

    Dissected how TIFAsomes branch into classical and alternative NF-κB outputs: TRAF6/TAK1 for classical and TRAF2-mediated cIAP1 displacement/degradation for alternative signaling.

    Evidence Reciprocal Co-IP, siRNA knockdown, IF, and immunoblotting in H. pylori-infected epithelial cells

    PMID:34328245

    Open questions at the time
    • Temporal coordination of the two branches within one condensate not resolved
    • Single-lab observation of the cIAP1 displacement step
  17. 2022 Medium

    Placed TIFA in intestinal homeostasis and cancer, showing the ALPK1/TIFA/TRAF6 axis drives barrier gene programs and pro-proliferative RSK/PRAS40 signaling requiring both Thr9 oligomerization and TRAF6 binding.

    Evidence CRISPR editing, structure-function mutagenesis (T9A, TRAF6-binding deletion), reporters, qPCR, and in vivo proliferation models

    PMID:35635239 PMID:36036242

    Open questions at the time
    • Whether commensal-driven and pathogen-driven TIFA signaling differ mechanistically unclear
    • In vivo causal contribution to tumorigenesis not fully established
  18. 2022 Medium

    Revealed a trafficking/antigen-presentation function in which TIFA binds calnexin and controls MHC-II compartment localization and invariant chain stability.

    Evidence siRNA knockdown, immunopeptidome analysis, Co-IP (TIFA-calnexin), IF, and CD4+ T cell assays

    PMID:36215666

    Open questions at the time
    • Relationship between this trafficking role and NF-κB signaling unresolved
    • Whether oligomerization is required not tested
  19. 2023 Medium

    Established that activated TIFA is turned over through both proteasomal and lysosomal routes via autophagy receptors, defining signal termination.

    Evidence Proteasome/lysosome inhibitors, Co-IP with polyubiquitin, optineurin, TAX1BP1, LAMP1, and infection time-course

    PMID:36965415

    Open questions at the time
    • The E3 ligase ubiquitinating TIFA not identified
    • Which degradation route dominates physiologically unclear
  20. 2024 Medium

    Showed TIFAsome formation is a liquid-liquid phase separation event requiring pThr9-FHA interaction and an intrinsically disordered region, with TRAF6 ubiquitination occurring inside condensates.

    Evidence Live-cell phase-separation imaging, chemical probe inhibition, in vitro ubiquitination, mutagenesis, and Co-IP

    PMID:38357697

    Open questions at the time
    • Material properties and regulation of condensate dissolution not defined
    • Single-lab characterization of the LLPS requirement
  21. 2024 High

    Provided the structural basis for TIFAB inhibition, showing it forms a pseudo-TIFA heterodimer with monomeric TIFA that lacks the phosphorylation site and TRAF6-binding motif.

    Evidence Crystallography of the TIFA/TIFAB heterodimer with SEC, in vitro binding, and NF-κB cell-based assays

    PMID:38442163

    Open questions at the time
    • How TIFAB levels are regulated to set the signaling threshold not addressed
  22. 2024 Medium

    Identified a non-NF-κB TIFA function in hepatocytes, where TIFA blocks PYK2 dimerization and recruits CBL for PYK2 degradation, dampening PYK2-JNK signaling.

    Evidence Hepatocyte-specific overexpression/deletion, Co-IP of the TIFA-PYK2-CBL complex, ubiquitination assays, and dietary rodent models

    PMID:39260704

    Open questions at the time
    • Whether this role depends on Thr9/oligomerization not tested
    • Tissue specificity versus the immune signaling role unclear
  23. 2025 Medium

    Refined TRAF2 engagement and degradation, defining a novel TIFA TRAF2-binding motif and showing ADP-heptose triggers TRAF2/TRAF6-dependent autophagic clearance of TIFA.

    Evidence Binding-motif mutagenesis, Co-IP, autophagy inhibitors, and TRAF2/TRAF6 knockdown

    PMID:40696502

    Open questions at the time
    • How TRAFs route TIFA to autophagy mechanistically unclear
    • Single-lab finding
  24. 2025 Medium

    Identified post-transcriptional control of TIFA, with METTL3/m6A-IGF2BP2 and NSUN3/m5C modifications stabilizing TIFA mRNA to amplify NF-κB/NLRP3-driven pyroptosis in kidney injury.

    Evidence MeRIP-seq/qPCR, RNA stability assays, conditional/knockdown models, Co-IP (IGF2BP2-TIFA mRNA), and in vivo AKI models

    PMID:39924309 PMID:41320097

    Open questions at the time
    • Whether these RNA modifications regulate TIFA in other tissues unknown
    • Crosstalk between mRNA stabilization and protein degradation not integrated

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the ALPK1/TIFA axis is integrated with other innate immune pathways such as STING and the integrated stress response, and how TIFAsome assembly/dissolution is quantitatively tuned in vivo, remains open.
  • Bidirectional ALPK1/TIFA-STING crosstalk reported only in a preprint without deep TIFA-specific dissection
  • No unified model linking condensate dynamics, degradation, and mRNA stabilization in setting signaling output

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 4 GO:0042393 histone binding 2
Localization
GO:0005829 cytosol 2 GO:0000228 nuclear chromosome 1 GO:0005634 nucleus 1 GO:0005794 Golgi apparatus 1
Pathway
R-HSA-162582 Signal Transduction 3 R-HSA-1643685 Disease 3 R-HSA-168256 Immune System 3
Partners
ALPK1CALNEXINIRAK1NEMOPYK2TIFABTRAF2TRAF6
Complex memberships
TIFA-TRAF2 complexTIFA-TRAF6 complexTIFA/TIFAB heterodimerTIFAsome

Evidence

Reading pass · 33 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 TIFA (T6BP) specifically associates with TRAF6 via its coiled-coil region interacting with the N-terminal ring finger and zinc finger domains of TRAF6. IL-1, but not TNF, induces TRAF6-T6BP complex formation in a ligand-dependent manner requiring IRAK; IRAK is not present in TRAF6-T6BP complexes. Yeast two-hybrid, co-immunoprecipitation Proceedings of the National Academy of Sciences of the United States of America Medium 10920205
2002 TIFA (T2BP) binds TRAF2 via the TRAF domain of TRAF2 and requires almost the entire TIFA protein for the interaction; overexpression of TIFA activates NF-κB and AP-1 in a dose-dependent manner without TNF stimulation. Mammalian two-hybrid, co-immunoprecipitation, NF-κB/AP-1 reporter assays Biochemical and biophysical research communications Medium 11798190
2003 TIFA bridges IRAK-1 and TRAF6 in IL-1 receptor signaling: TIFA associates with TRAF6 constitutively and with IRAK-1 in an IL-1-stimulation-dependent manner; mutations abolishing TRAF6 binding or FHA domain phosphopeptide binding prevent NF-κB and JNK activation; overexpressed TIFA significantly enhances the IRAK-1/TRAF6 interaction. Co-immunoprecipitation of endogenous proteins, transient transfection with NF-κB/JNK reporter assays, site-directed mutagenesis The Journal of biological chemistry High 12566447
2004 TIFA activates IKK by promoting oligomerization and K63-linked polyubiquitination of TRAF6, which then activates TAK1 and IKK through a proteasome-independent mechanism. Only high-molecular-weight oligomeric forms of TIFA (not TRAF6-binding-defective mutant) activate IKK. Demonstrated in an in vitro reconstitution system with purified TIFA, TRAF6, TAK1 kinase complex, and Ubc13-Uev1A. In vitro reconstitution with purified proteins, cell-free IKK activation assay, in vitro ubiquitination assay, mutagenesis Proceedings of the National Academy of Sciences of the United States of America High 15492226
2004 TIFAB, a TIFA homolog, inhibits TIFA-mediated NF-κB activation by binding TIFA. TIFAB does not associate with TRAF family members but binds TIFA and increases the amount of TRAF6 co-precipitated with TIFA, suggesting TIFAB induces a conformational change in TIFA that inhibits TRAF6 activation. Co-immunoprecipitation, NF-κB reporter assay Biochemical and biophysical research communications Medium 15047173
2006 ZCCHC11, a zinc finger protein, interacts with TIFA via affinity purification from macrophage phosphoproteins. Upon LPS stimulation, ZCCHC11L translocates from nucleus to cytoplasm and binds TIFA. ZCCHC11 functions as a negative regulator of TLR-mediated NF-κB activation downstream of TIFA-TRAF6. GST pulldown/affinity purification with mass spectrometry, co-immunoprecipitation, siRNA knockdown, NF-κB reporter assay Biochemical and biophysical research communications Medium 16643855
2007 TIFA (T6BP) is a myosin VI binding partner; the interaction was confirmed in vitro and in vivo with binding sites mapped on each protein. T6BP and myosin VI localize to the trans-Golgi complex and perinuclear vesicles. RNAi knockdown of T6BP reduces membrane ruffling, increases stress fibers and focal adhesions, upregulates constitutive secretion, and inhibits NF-κB activation. Yeast two-hybrid, in vitro binding, co-immunoprecipitation, immunofluorescence, electron microscopy, RNAi knockdown with functional assays Journal of cell science High 17635994
2012 TIFA undergoes phosphorylation at Thr9, and this phosphorylated Thr9 (pT9) binds intramolecularly with the FHA domain of TIFA from a different dimer, driving TIFA oligomerization and NF-κB activation. Unphosphorylated TIFA exists as an intrinsic dimer. Silencing endogenous TIFA attenuates TNF-α-mediated downstream NF-κB signaling. Biochemical analysis, phosphorylation mapping, TIFA oligomerization assays, siRNA knockdown, NF-κB reporter assay Molecular and cellular biology High 22566686
2013 TIFA upregulation during hypoxia-reoxygenation is TLR4- and MyD88-dependent. TIFA associates with TRAF6 constitutively and with IRAK-1 only after hypoxia-reoxygenation. siRNA-mediated TIFA knockdown reduces NF-κB activation and HMGB1 upregulation/release after hypoxia-reoxygenation. Co-immunoprecipitation, siRNA knockdown, EMSA, Western blotting, TLR4(-/-) and MyD88(-/-) cells Free radical biology & medicine Medium 23722163
2015 Cytosolic detection of the bacterial metabolite heptose-1,7-bisphosphate (HBP) activates NF-κB through a TIFA-dependent signaling axis mediated by phosphorylation-dependent oligomerization of TIFA. Identified by genome-wide RNAi screen and validated in cell-based assays. Genome-wide RNAi screen, cell-based NF-κB activation assay, TIFA phosphorylation and oligomerization assays Science (New York, N.Y.) High 26068852
2015 Crystal structure of TIFA (residues 1-150) and its complex with the N-terminal pThr9 peptide (residues 1-15) reveals: TIFA forms an intrinsic dimer, has an extra β-strand in the FHA structure, and pThr9-FHA domain interaction occurs only between different dimers (not within a dimer), providing the structural mechanism for TIFA oligomerization. X-ray crystallography, size-exclusion chromatography, mutagenesis, functional NF-κB assays Biochemistry High 26389808
2016 Aurora A kinase phosphorylates TIFA at Thr9, triggering NF-κB survival pathway activation in AML cells. TIFA silencing decreases leukemic cell growth and chemoresistance by downregulating Bcl-2 and Bcl-XL. In vivo delivery of TIFA-inhibitory fragments potentiated myeloblast clearance in xenograft mice. siRNA knockdown, in vivo xenograft, kinase assay (Aurora A phosphorylation of TIFA Thr9), Western blotting, IC50 assays Cancer research Medium 28069801
2016 TIFA mediates both signal 1 (priming via NF-κB) and signal 2 (activation) of the NLRP3 inflammasome in vascular endothelial cells. STEROL regulatory element-binding protein 2 transactivates TIFA expression (signal 1). Akt phosphorylates TIFA at Thr9 (signal 2), enabling TIFA-TIFA homophilic oligomerization and interaction with caspase-1 for higher-order NLRP3 assembly. Transcription factor binding/reporter assay, Akt kinase assay, co-immunoprecipitation (TIFA-caspase-1), siRNA knockdown, in vitro and in vivo atheroprone flow models Proceedings of the National Academy of Sciences of the United States of America Medium 27965388
2017 ALPK1 kinase phosphorylates TIFA at Thr9 in response to H. pylori-delivered βHBP; this triggers TIFAsome formation (large TIFA oligomeric complexes including TRAF2) and NF-κB activation. ALPK1 knockout and CRISPR/Cas9 ablation abrogate TIFA phosphorylation and TIFAsome formation. The pathway is independent of CagA translocation. CRISPR/Cas9 knockout, RNAi, immunofluorescence microscopy, immunoblotting, mass spectrometry, recombinant protein technology Cell reports High 28877472
2017 TIFA threonine 9 phosphorylation and FHA domain are required for TIFA oligomerization in both infected and bystander cells in response to Shigella and Salmonella infection. ALPK1 is the critical kinase responsible for TIFA oligomerization, TRAF6 oligomerization, and IL-8/NF-κB activation. Connexin-dependent cell-cell communication enables bystander cell activation via this pathway. Genome-wide RNAi screen, mutagenesis (Thr9), siRNA knockdown, NF-κB/IL-8 reporter assays, TIFA oligomerization assays PLoS pathogens High 28222186
2017 During Shigella infection, TIFA senses HBP released during intracellular bacterial replication (after NOD1-mediated entry), assembling into large signaling complexes; IECs lacking TIFA cannot discriminate between proliferating and stagnant intracellular bacteria, establishing TIFA as a rheostat for intracellular bacterial replication that escalates immune response proportionally to replication rate. TIFA knockout cells, siRNA knockdown, NF-κB activation assays, bacterial replication quantification, immunofluorescence Cell reports High 28514661
2017 HBP is delivered to host gastric epithelial cells via the cag-T4SS where it activates TIFA-dependent NF-κB signaling independently of NOD1; TIFA activation precedes NOD1 activation during H. pylori infection. CagA contributes to NF-κB downstream of TIFA and NOD1. siRNA knockdown, bacterial mutant strains (cagPAI-deficient), NF-κB reporter assays, time-course analyses mBio High 28811347
2018 Direct binding between TIFA C-terminal peptide and the TRAF domain of TRAF6 was demonstrated in vitro; crystal structures of TRAF domain complexed with WT and TIFA S174Q/M179D mutant peptides show salt-bridge formation between TIFA residues 177-181 and TRAF6 binding pocket. The double mutant shows enhanced binding to endogenous full-length TRAF6. X-ray crystallography, in vitro binding assay, co-immunoprecipitation with mutants Chembiochem : a European journal of chemical biology High 30378729
2018 Upon genotoxic stress, TIFA undergoes nuclear translocation and accumulates on damaged chromatin. DNA damage induces TIFA phosphorylation at Thr9; this pThr9-FHA domain interaction drives enrichment on damaged chromatin and NF-κB activation. TIFA, together with TRAF2, stimulates NEMO ubiquitination to relay DNA damage signals. TRAF2 knockdown suppresses TIFA-enhanced NEMO ubiquitination; unphosphorylatable T9A TIFA mutant has minimal effect on NEMO polyubiquitination. Subcellular fractionation, immunofluorescence, mutagenesis (Thr9), co-immunoprecipitation, ubiquitination assay, siRNA knockdown The Journal of biological chemistry Medium 29581234
2020 Crystal structures of mouse TIFA and Thr9Asp/Thr9Glu phosphomimetic mutants confirm dimer formation similar to human TIFA. Size-exclusion chromatography modeling suggests a higher-order TIFA-TRAF6 signaling complex. Small-angle X-ray scattering confirms the dimer as the solution structure. X-ray crystallography, size-exclusion chromatography, small-angle X-ray scattering Scientific reports High 32198460
2020 H. pylori-induced DNA double-strand breaks occur co-transcriptionally in S-phase cells activating NF-κB via β-ADP-heptose/ALPK1/TIFA signaling. R-loops (RNA/DNA hybrids) form during S-phase as a consequence of TIFA/NF-κB signaling, and replication stress/DNA damage depend on these R-loops. DNA damage requires the H. pylori RfaE enzyme and Cag pathogenicity island. CRISPR/Cas9 knockout, gastric organoid primary cells, R-loop detection (immunofluorescence/S9.6 antibody), DNA fiber assay, bacterial mutant strains Nature communications Medium 33037203
2021 TIFA interacts with both TRAF6 and TRAF2 in H. pylori-infected gastric epithelial cells, forming TIFAsomes. The TIFA/TRAF6 interaction enables TAK1 binding, leading to classical NF-κB activation. The TIFA/TRAF2 interaction causes transient displacement and proteasomal degradation of cIAP1 from TRAF2, facilitating alternative NF-κB pathway activation. Co-immunoprecipitation, siRNA knockdown, immunofluorescence, immunoblotting EMBO reports Medium 34328245
2022 TIFA expression is essential for intestinal epithelial cell responsiveness to ADP-heptose from Akkermansia muciniphila; TIFA/ALPK1/TRAF6-dependent NF-κB activation drives MUC2, BIRC3, and TNFAIP3 gene expression for intestinal barrier maintenance. Genetic editing tools confirm the pathway. CRISPR/Cas9 gene editing, NF-κB reporter assay, siRNA, pharmacological inhibitors, qPCR Gut microbes Medium 36036242
2022 TIFA-T9A (oligomerization site mutation) and TIFA-D6 (TRAF6-binding site deletion) mutants abolish TIFA-mediated CRC cell proliferation enhancement, confirming that both Thr9 phosphorylation-dependent oligomerization and TRAF6 binding are required for TIFA's pro-proliferative function. RSK and PRAS40 activation downstream of TIFA drives CRC progression. Site-directed mutagenesis, siRNA knockdown, ectopic expression, in vitro/in vivo proliferation assays, Western blotting Cancer science Medium 35635239
2022 TIFA (T6BP) influences MHC-II-restricted endogenous antigen presentation. T6BP silencing causes mislocalization of MHC-II loading compartments and rapid degradation of the invariant chain (CD74) without affecting MHC-II expression or internalization. Calnexin is identified as a T6BP binding partner (via its cytosolic tail), and calnexin silencing replicates T6BP knockdown phenotypes. siRNA knockdown, immunopeptidome analysis of MHC-II molecules, co-immunoprecipitation (T6BP-calnexin), immunofluorescence, CD4+ T cell activation assay EMBO reports Medium 36215666
2023 H. pylori promotes TIFA degradation via both proteasomal and lysosomal pathways following ALPK1-dependent activation. TIFA interacts with polyubiquitin, optineurin, TAX1BP1, and LAMP1 after H. pylori infection; TRAF2, TRAF6, TAK1, and NEMO are not required for TIFA degradation. Proteasome/lysosome inhibitor treatment, co-immunoprecipitation, immunoblotting, H. pylori infection time-course European journal of cell biology Medium 36965415
2024 ADP-heptose (ADP-Hep) recognition by ALPK1 induces liquid-liquid phase separation (LLPS) of TIFA, driven by ALPK1 phosphorylation of pT9, the pT9-FHA domain interaction, and an intrinsically disordered region. TRAF6 is recruited into TIFA condensates, enabling K63-linked polyubiquitin chain synthesis within condensates. A chemical probe (compound 22) confirmed that ALPK1-TIFA-TRAF6 pathway activation requires TIFA phase separation. Live-cell imaging of phase separation, chemical probe inhibition, in vitro ubiquitination assay, mutagenesis, Co-IP Research (Washington, D.C.) Medium 38357697
2024 TIFAB forms a stable heterodimer with monomeric TIFA (not with the TIFA dimer), inhibiting TIFA dimer formation and suppressing TIFA-TRAF6 signaling. Crystal structure of the TIFA/TIFAB heterodimer shows it forms a 'pseudo-TIFA dimer' lacking the phosphorylation site and TRAF6-binding motif, preventing formation of the ordered phosphorylated TIFA oligomer required for NF-κB activation. X-ray crystallography, biochemical assays (SEC, in vitro binding), cell-based NF-κB assays, co-immunoprecipitation Proceedings of the National Academy of Sciences of the United States of America High 38442163
2024 TIFA (T6BP) directly targets PYK2 and prevents N-terminal FERM domain-triggered PYK2 dimerization, disrupting PYK2-JNK signaling. TIFA also recruits the E3 ubiquitin ligase CBL to form a complex with PYK2, leading to CBL-mediated PYK2 degradation. Hepatocyte-specific overexpression/deletion, co-immunoprecipitation (TIFA-PYK2-CBL complex), ubiquitination assay, in vivo dietary rodent models Journal of hepatology Medium 39260704
2025 TRAF2 interacts with TIFA via two conserved sequence motifs, one of which (Pro159-Xaa-Xaa-Glu162) is novel. ADP-heptose induces TIFA degradation by autophagy, and both TRAF2 and TRAF6 contribute to this autophagic degradation of TIFA. Mutagenesis of TIFA binding motifs, co-immunoprecipitation, autophagy inhibitor assays, TRAF2/TRAF6 knockdown FEBS letters Medium 40696502
2025 METTL3 mediates m6A modification of TIFA mRNA, which is recognized by IGF2BP2 to enhance TIFA mRNA stability, thereby upregulating TIFA protein; elevated TIFA promotes NLRP3 transcription via NF-κB signaling, activating NLRP3 inflammasome and Caspase-1 to drive pyroptosis in tubular epithelial cells during AKI. MeRIP-seq, RNA sequencing, METTL3 conditional knockout mice, siRNA knockdown, co-immunoprecipitation (IGF2BP2-TIFA mRNA), NLRP3 inflammasome/pyroptosis assays Free radical biology & medicine Medium 41320097
2025 NSUN3 stabilizes TIFA mRNA through m5C methylation, increasing TIFA protein expression; NSUN3 knockdown reduces TIFA expression, alleviates LPS-induced HK-2 cell injury, and reduces SA-AKI in mice. methylation RNA immunoprecipitation-qPCR, actinomycin D mRNA stability assay, siRNA knockdown, in vivo CLP model Clinical and experimental pharmacology & physiology Medium 39924309
2025 ALPK1 activation enhances STING pathway outputs including LC3B lipidation and NLRP3 inflammasome activation; conversely, STING activation increases ALPK1 protein expression and triggers TIFA Thr9 phosphorylation, demonstrating bidirectional crosstalk between the ALPK1/TIFA and STING pathways. ALPK1 signaling also activates eIF2α (integrated stress response). Cell-based pathway activation assays, Western blotting (TIFA Thr9 phosphorylation), LC3B lipidation assay, NLRP3 inflammasome assay bioRxivpreprint Low bio_10.1101_2025.06.30.662363

Source papers

Stage 0 corpus · 46 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2017 ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System. Cell reports 148 28877472
2015 INNATE IMMUNITY. Cytosolic detection of the bacterial metabolite HBP activates TIFA-dependent innate immunity. Science (New York, N.Y.) 129 26068852
2017 TIFA Signaling in Gastric Epithelial Cells Initiates the cag Type 4 Secretion System-Dependent Innate Immune Response to Helicobacter pylori Infection. mBio 118 28811347
2004 TIFA activates IkappaB kinase (IKK) by promoting oligomerization and ubiquitination of TRAF6. Proceedings of the National Academy of Sciences of the United States of America 116 15492226
2017 ALPK1 controls TIFA/TRAF6-dependent innate immunity against heptose-1,7-bisphosphate of gram-negative bacteria. PLoS pathogens 106 28222186
2020 The ALPK1/TIFA/NF-κB axis links a bacterial carcinogen to R-loop-induced replication stress. Nature communications 91 33037203
2007 T6BP and NDP52 are myosin VI binding partners with potential roles in cytokine signalling and cell adhesion. Journal of cell science 80 17635994
2003 Identification of TIFA as an adapter protein that links tumor necrosis factor receptor-associated factor 6 (TRAF6) to interleukin-1 (IL-1) receptor-associated kinase-1 (IRAK-1) in IL-1 receptor signaling. The Journal of biological chemistry 75 12566447
2022 Akkermansia muciniphila upregulates genes involved in maintaining the intestinal barrier function via ADP-heptose-dependent activation of the ALPK1/TIFA pathway. Gut microbes 68 36036242
2000 T6BP, a TRAF6-interacting protein involved in IL-1 signaling. Proceedings of the National Academy of Sciences of the United States of America 56 10920205
2002 T2BP, a novel TRAF2 binding protein, can activate NF-kappaB and AP-1 without TNF stimulation. Biochemical and biophysical research communications 55 11798190
2023 Fusobacterium nucleatum promotes inflammatory and anti-apoptotic responses in colorectal cancer cells via ADP-heptose release and ALPK1/TIFA axis activation. Gut microbes 53 38126163
2017 Innate Recognition of Intracellular Bacterial Growth Is Driven by the TIFA-Dependent Cytosolic Surveillance Pathway. Cell reports 53 28514661
2016 Aurora A and NF-κB Survival Pathway Drive Chemoresistance in Acute Myeloid Leukemia via the TRAF-Interacting Protein TIFA. Cancer research 48 28069801
2016 TIFA as a crucial mediator for NLRP3 inflammasome. Proceedings of the National Academy of Sciences of the United States of America 44 27965388
2021 TIFA has dual functions in Helicobacter pylori-induced classical and alternative NF-κB pathways. EMBO reports 42 34328245
2006 A novel Zinc finger protein, ZCCHC11, interacts with TIFA and modulates TLR signaling. Biochemical and biophysical research communications 41 16643855
2012 Intermolecular binding between TIFA-FHA and TIFA-pT mediates tumor necrosis factor alpha stimulation and NF-κB activation. Molecular and cellular biology 40 22566686
2016 TIFA suppresses hepatocellular carcinoma progression via MALT1-dependent and -independent signaling pathways. Signal transduction and targeted therapy 37 29263897
2015 TIFA, an inflammatory signaling adaptor, is tumor suppressive for liver cancer. Oncogenesis 32 26501855
2013 TIFA upregulation after hypoxia-reoxygenation is TLR4- and MyD88-dependent and associated with HMGB1 upregulation and release. Free radical biology & medicine 31 23722163
2015 Uncovering the Mechanism of Forkhead-Associated Domain-Mediated TIFA Oligomerization That Plays a Central Role in Immune Responses. Biochemistry 29 26389808
2020 TIFA and TIFAB: FHA-domain proteins involved in inflammation, hematopoiesis, and disease. Experimental hematology 24 32910997
2004 TIFAB inhibits TIFA, TRAF-interacting protein with a forkhead-associated domain. Biochemical and biophysical research communications 20 15047173
2022 The potential biomarker TIFA regulates pyroptosis in sepsis-induced acute kidney injury. International immunopharmacology 19 36586274
2019 Role of NOD1 and ALPK1/TIFA Signalling in Innate Immunity Against Helicobacter pylori Infection. Current topics in microbiology and immunology 19 31123889
2018 TRAF-interacting protein with forkhead-associated domain (TIFA) transduces DNA damage-induced activation of NF-κB. The Journal of biological chemistry 15 29581234
2018 Binding and Enhanced Binding between Key Immunity Proteins TRAF6 and TIFA. Chembiochem : a European journal of chemical biology 14 30378729
2018 TIFA Promotes Cell Survival and Migration in Lung Adenocarcinoma. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 13 29975933
2022 TIFA promotes colorectal cancer cell proliferation in an RSK- and PRAS40-dependent manner. Cancer science 12 35635239
2022 The Autophagy Receptor TAX1BP1 (T6BP) improves antigen presentation by MHC-II molecules. EMBO reports 12 36215666
2024 Targeting PYK2 with heterobifunctional T6BP helps mitigate MASLD and MASH-HCC progression. Journal of hepatology 10 39260704
2020 Structural analysis of TIFA: Insight into TIFA-dependent signal transduction in innate immunity. Scientific reports 10 32198460
2020 Nonhydrolyzable Heptose Bis- and Monophosphate Analogues Modulate Pro-inflammatory TIFA-NF-κB Signaling. Chembiochem : a European journal of chemical biology 9 32452604
2025 NSUN3 Aggravates Sepsis-Associated Acute Kidney Injury by Stabilising TIFA mRNA Through m5C. Clinical and experimental pharmacology & physiology 8 39924309
2024 ADP-Hep-Induced Liquid Phase Condensation of TIFA-TRAF6 Activates ALPK1/TIFA-Dependent Innate Immune Responses. Research (Washington, D.C.) 8 38357697
2023 Helicobacter pylori regulates TIFA turnover in gastric epithelial cells. European journal of cell biology 7 36965415
2024 TIFAB regulates the TIFA-TRAF6 signaling pathway involved in innate immunity by forming a heterodimer complex with TIFA. Proceedings of the National Academy of Sciences of the United States of America 6 38442163
2005 Identification and characterization of Xenopus laevis homologs of mammalian TRAF6 and its binding protein TIFA. Gene 6 16023795
2024 IFN-γ licenses normal and pathogenic ALPK1/TIFA pathway in human monocytes. iScience 4 39868044
2023 Inducing a Proinflammatory Response with Bioengineered Yeast Vacuoles with TLR2-Binding Peptides (VacT2BP) as a Drug Carrier for Daunorubicin Delivery. ACS applied materials & interfaces 4 37615983
2023 TIFA contributes to periodontitis in diabetic mice via activating the NF‑κB signaling pathway. Molecular medicine reports 4 38099344
2025 TIFA renders intestinal epithelial cells responsive to microbial ADP-heptose and drives colonic inflammation in mice. Mucosal immunology 3 39842611
2024 TIFA enhances glycolysis through E2F1 and promotes the progression of glioma. Cellular signalling 2 39481822
2025 METTL3-mediated m6A modification of TIFA mRNA promotes tubular cell pyroptosis in acute kidney injury. Free radical biology & medicine 1 41320097
2025 TRAF2 binds to TIFA via a novel motif and contributes to its autophagic degradation. FEBS letters 0 40696502

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