Affinage

TRIM4

E3 ubiquitin-protein ligase TRIM4 · UniProt Q9C037

Length
500 aa
Mass
57.5 kDa
Annotated
2026-06-10
14 papers in source corpus 12 papers cited in narrative 12 extracted findings
Cross-family judge vs UniProt: tie faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

TRIM4 is a RING-domain E3 ubiquitin ligase that governs both antiviral innate immunity and proteostasis by directing distinct polyubiquitin linkages onto multiple substrates (PMID:24755855, PMID:35843886). In the RIG-I/MDA5 sensing pathway it catalyzes K63-linked polyubiquitination of RIG-I and MDA5, driving IRF3/NF-κB activation and type I interferon induction (PMID:24755855, PMID:37956198). This antiviral arm is targeted by pathogens: SARS-CoV-2 Nsp8 bridges MDA5 and TRIM4 to block the modification and suppress IFN (PMID:37956198), while TRIM4 directly degrades viral proteins including influenza A NP and PRRSV NSP2 through the proteasome to restrict replication (PMID:35637527, PMID:37823597). In its degradative mode TRIM4 catalyzes K48-linked ubiquitination of cellular substrates at defined lysines—SET at K150/K172, engaging the p53/PP2A/ESR1 axis, and TPL2 at K415/K439, controlling GSK3β and β-catenin/Wnt signaling—and additionally degrades hnRNPDL and ABTB1 to influence CDK4/6-inhibitor sensitivity and CDK1-dependent G2/M progression (PMID:35843886, PMID:39178114, PMID:39643799, PMID:42034143). TRIM4 substrate selection is achieved through discrete domains (RING and B-box for SET and hnRNPDL, SPRY for TRPM8), and its own abundance is controlled by a KRAS–RNF185–TRIM21 stabilization circuit and by ABTB1-driven turnover (PMID:35843886, PMID:33037615, PMID:39178114, PMID:37823597). Beyond ubiquitination, TRIM4 forms cytoplasmic speckles that transiently engage mitochondria and binds PRX1 to sensitize cells to H2O2-induced death (PMID:26524401), and it negatively regulates the TRPM8 ion channel via K63-ubiquitination at K423 (PMID:33037615).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 2014 Medium

    Established TRIM4 as a positive regulator of antiviral signaling, answering whether this TRIM acts on a nucleic-acid sensor rather than acting only as a degradative ligase.

    Evidence Reciprocal Co-IP plus overexpression/knockdown with IRF3, NF-κB and IFN-β reporters in response to viral RNA

    PMID:24755855

    Open questions at the time
    • Direct in vitro reconstitution of RIG-I K63-ubiquitination not shown
    • Ubiquitin-acceptor lysines on RIG-I not mapped
  2. 2015 Medium

    Defined a ubiquitination-independent role linking TRIM4 to mitochondrial oxidative-stress responses and identified PRX1 as a partner mediating death sensitization.

    Evidence Subcellular imaging, mitochondrial ROS/membrane-potential assays, Co-IP/MS for PRX1 in H2O2-treated cells

    PMID:26524401

    Open questions at the time
    • Whether PRX1 is a ubiquitination substrate unresolved
    • Molecular trigger of mitochondrial speckle association unknown
  3. 2020 High

    Showed TRIM4 regulates ion-channel surface expression, broadening its substrate repertoire to a membrane channel via the SPRY domain and a defined acceptor lysine.

    Evidence SPRY-domain Co-IP mapping, patch-clamp, surface biotinylation, K423 mutagenesis and UBA1 knockdown

    PMID:33037615

    Open questions at the time
    • Physiological context of TRPM8 regulation in cold/pain signaling untested
    • Whether K63-ubiquitination drives internalization mechanistically not dissected
  4. 2022 High

    Demonstrated a K48-linked degradative function with site-specific resolution, connecting TRIM4 to the SET/p53/PP2A/ESR1 axis in cancer.

    Evidence Domain mapping, in vitro/cell ubiquitination with K150/K172 mutagenesis, xenografts

    PMID:35843886

    Open questions at the time
    • Tissue specificity of the SET-ERα axis not defined
    • Upstream signals regulating TRIM4-SET engagement unknown
  5. 2022 Medium

    Extended the antiviral degradative role to a non-human virus, showing TRIM4 directly degrades a viral protein to restrict replication.

    Evidence Knockdown/overexpression PRRSV replication assays, ubiquitination assays, MG132 rescue

    PMID:35637527

    Open questions at the time
    • NSP2 acceptor lysines not mapped
    • Ubiquitin linkage type on NSP2 not determined
  6. 2022 Low

    Confirmed conservation of TRIM4's positive IFN-pathway role across primates.

    Evidence Overexpression/siRNA with IFN-β/NF-κB/ISRE reporters and qRT-PCR in Macaca mulatta cells

    PMID:35242838

    Open questions at the time
    • No mechanistic dissection of direct substrate interactions in this system
    • Human relevance inferred not tested here
  7. 2023 Medium

    Identified MDA5 as a second sensor substrate and revealed a viral evasion strategy targeting TRIM4-MDA5 ubiquitination.

    Evidence Co-IP, ubiquitination assays, IFN-β/NF-κB/ISRE reporters; SARS-CoV-2 Nsp8 binding studies

    PMID:37956198

    Open questions at the time
    • MDA5 acceptor lysines not mapped
    • Stoichiometry of the Nsp8-MDA5-TRIM4 ternary interaction unresolved
  8. 2023 Medium

    Showed TRIM4 restricts influenza by degrading NP and revealed ABTB1 as a regulator of TRIM4 stability.

    Evidence IP-MS, viral replication assays, ubiquitination assays, proteasome rescue

    PMID:37823597

    Open questions at the time
    • NP degradation lysines not defined
    • Whether ABTB1 directly ubiquitinates TRIM4 not established here
  9. 2024 High

    Placed TRIM4 within an oncogenic KRAS circuit, defining TPL2 as a substrate and a KRAS-RNF185-TRIM21 cascade controlling TRIM4 levels.

    Evidence BioID, Co-IP, K415/K439 mutagenesis, genetic perturbation of KRAS/TRIM21/RNF185

    PMID:39178114

    Open questions at the time
    • Direct TRIM21-mediated TRIM4 ubiquitination sites not mapped
    • Generality of the KRAS circuit across tumor types untested
  10. 2024 Medium

    Linked TRIM4 to CDK4/6-inhibitor sensitivity through K48-degradation of hnRNPDL and its downstream control of CDKN2C splicing.

    Evidence Co-IP, GST pull-down domain mapping, ubiquitination assays, RIP, in vivo siTRIM4

    PMID:39643799

    Open questions at the time
    • hnRNPDL acceptor lysines not mapped
    • Direct vs indirect effects on CDKN2C splicing not fully separated
  11. 2024 Medium

    Demonstrated atypical multi-linkage ubiquitination of ABTB1 by TRIM4, coupling TRIM4 to CDK1 stabilization and G2/M progression in glioblastoma.

    Evidence Co-IP, domain/lysine-specific ubiquitination assays, cell-cycle flow cytometry, xenografts

    PMID:42034143

    Open questions at the time
    • Functional consequence of each individual linkage type unresolved
    • Reciprocal TRIM4/ABTB1 regulation across contexts not reconciled with the influenza ABTB1 finding
  12. 2024 Medium

    Revealed a drug-induced molecular-glue/neddylation mechanism, showing TRIM4 can be redirected to degrade CORO1A in the presence of Aurovertin B.

    Evidence Co-IP, MS, SPPIER, phenotypic drug screen with multi-omics, patient-derived organoids

    PMID:39629122

    Open questions at the time
    • Neddylation versus ubiquitination contributions not biochemically separated
    • Endogenous (drug-independent) relevance of CORO1A targeting unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How TRIM4 selects between K48-degradative and K63/signaling outputs on different substrates, and what upstream signals partition these activities, remains undefined.
  • No structural model of substrate engagement
  • Linkage-determining cofactors (E2 enzymes) largely unidentified
  • Cell-type-specific regulation of competing functions unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016874 ligase activity 7 GO:0140096 catalytic activity, acting on a protein 5 GO:0031386 protein tag activity 3
Localization
GO:0005739 mitochondrion 1 GO:0005829 cytosol 1
Pathway
R-HSA-168256 Immune System 4 R-HSA-392499 Metabolism of proteins 4 R-HSA-162582 Signal Transduction 3 R-HSA-1640170 Cell Cycle 2

Evidence

Reading pass · 12 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2014 TRIM4 associates with RIG-I and targets it for K63-linked polyubiquitination, positively regulating RIG-I-mediated type I interferon induction and activation of IRF3 and NF-κB in response to viral RNA. Co-immunoprecipitation, overexpression and knockdown with reporter assays for IRF3, NF-κB, and IFN-β activation Journal of molecular cell biology Medium 24755855
2015 TRIM4 forms cytoplasmic speckle-like structures that transiently interact with mitochondria, induces mitochondrial aggregation and increased mitochondrial ROS in the presence of H2O2, potentiates loss of mitochondrial transmembrane potential and cytochrome c release, and interacts with peroxiredoxin 1 (PRX1); this interaction is critical for H2O2-induced cell death sensitization. Subcellular localization imaging, overexpression/knockdown cell death assays, Co-immunoprecipitation/mass spectrometry for PRX1 interaction, mitochondrial ROS and membrane potential measurements Free radical biology & medicine Medium 26524401
2022 TRIM4 interacts with SET via its RING and B-box domains (and SET's carboxyl terminus) and catalyzes K48-linked polyubiquitination of SET at K150 and K172, promoting proteasomal degradation of SET, which releases p53 and PP2A to promote ESR1 transcription and enhance ER-α expression. Co-immunoprecipitation, domain-mapping experiments, in vitro/cell-based ubiquitination assays with site-specific mutagenesis (K150/K172), in vivo xenograft models Advanced science (Weinheim, Baden-Wurttemberg, Germany) High 35843886
2023 TRIM4 acts as an E3 ubiquitin ligase for MDA5, mediating K63-linked polyubiquitination of MDA5 to activate antiviral innate immune responses; SARS-CoV-2 Nsp8 binds both MDA5 and TRIM4 to impair this ubiquitination and suppress IFN induction. Co-immunoprecipitation, ubiquitination assays, reporter assays for IFN-β/NF-κB/ISRE, overexpression/knockdown functional studies PLoS pathogens Medium 37956198
2020 TRIM4 interacts with TRPM8 via its SPRY domain and promotes K63-linked ubiquitination of TRPM8 at lysine K423, reducing TRPM8 cell-surface expression and negatively regulating TRPM8-mediated currents; this regulation requires the E1 enzyme UBA1. Co-immunoprecipitation (SPRY domain mapping), patch-clamp electrophysiology, cell-surface biotinylation, site-directed mutagenesis (K423), UBA1 knockdown Journal of cellular physiology High 33037615
2022 TRIM4 inhibits PRRSV replication by ubiquitinating and degrading the viral NSP2 protein via the proteasome; TRIM4 knockdown increases virus titers and MG132 treatment blocks TRIM4-driven NSP2 degradation. Overexpression/siRNA knockdown viral replication assays, ubiquitination assays, proteasome inhibitor (MG132) rescue experiments BMC veterinary research Medium 35637527
2023 TRIM4 uses its E3 ubiquitin ligase activity to degrade influenza A virus NP protein (within incoming vRNP and newly synthesized NP), thereby inhibiting IAV replication; ABTB1 counteracts this by interacting with TRIM4 and promoting TRIM4 degradation through the proteasome. Immunoprecipitation and mass spectrometry, overexpression/knockdown viral replication assays, ubiquitination assays, proteasome inhibitor rescue Emerging microbes & infections Medium 37823597
2024 TRIM4 is the E3 ligase that binds and degrades TPL2 kinase via polyubiquitination at lysines K415 and K439; TRIM4 itself is stabilized by E3 ligase TRIM21, which is regulated by KRAS—mutant KRAS recruits RNF185 to degrade TRIM21, thereby destabilizing TRIM4 and stabilizing TPL2, leading to GSK3β phosphorylation/degradation and β-catenin/Wnt pathway activation. Proximity-dependent biotin identification (BioID), Co-immunoprecipitation, ubiquitination assays with site-specific mutagenesis (K415, K439), genetic perturbation of KRAS/TRIM21/RNF185 Cell reports High 39178114
2024 TRIM4 binds hnRNPDL via its RING and B-box domains and promotes K48-linked ubiquitination and proteasomal degradation of hnRNPDL; hnRNPDL in turn binds CDKN2C isoform 2 mRNA and suppresses its expression via alternative splicing, thereby weakening sensitivity to CDK4/6 inhibitors. Co-immunoprecipitation, GST pull-down (domain mapping), ubiquitination assays, RIP (RNA immunoprecipitation) assay, in vivo siTRIM4 experiments Frontiers of medicine Medium 39643799
2024 TRIM4 promotes ABTB1 degradation through K6, K27, K29, and K33-linked ubiquitination (mediated by the TRIM4 53–500 aa region), thereby preventing ABTB1 from ubiquitinating CDK1 (via K27-linked ubiquitination) and stabilizing CDK1 to promote G2/M progression in glioblastoma. Co-immunoprecipitation, ubiquitination assays with domain/lysine-specific mutagenesis, protein turnover assays, flow cytometry (cell cycle), xenograft tumor models International journal of biological macromolecules Medium 42034143
2024 TRIM4 acts as an E3 ligase (atypical neddylation pathway) facilitating the neddylation and proteasomal degradation of CORO1A when bound to the small molecule Aurovertin B, functioning as a molecular glue mechanism; TRIM4-CORO1A interaction was confirmed by Co-IP, mass spectrometry, and SPPIER assay. Co-immunoprecipitation, mass spectrometry, SPPIER (separation of phases-based protein interaction reporter), phenotypic drug screen with multi-omics, patient-derived organoids Theranostics Medium 39629122
2022 Macaca mulatta TRIM4 overexpression upregulates IFN-alpha, IFN-beta, RIG-I, MAVS, IRF3, IRF7, and antiviral gene expression, and increases IFN-β, NF-κB, and ISRE reporter activity; siRNA knockdown of TRIM4 downregulates the IFN pathway, confirming a conserved positive regulatory role in the interferon signaling pathway. Overexpression and siRNA knockdown with luciferase reporter assays and qRT-PCR in primate cells Frontiers in veterinary science Low 35242838

Source papers

Stage 0 corpus · 14 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 TRIM4 modulates type I interferon induction and cellular antiviral response by targeting RIG-I for K63-linked ubiquitination. Journal of molecular cell biology 185 24755855
2015 TRIM4; a novel mitochondrial interacting RING E3 ligase, sensitizes the cells to hydrogen peroxide (H2O2) induced cell death. Free radical biology & medicine 34 26524401
2023 SARS-CoV-2 Nsp8 suppresses MDA5 antiviral immune responses by impairing TRIM4-mediated K63-linked polyubiquitination. PLoS pathogens 27 37956198
2022 The E3 Ligase TRIM4 Facilitates SET Ubiquitin-Mediated Degradation to Enhance ER-α Action in Breast Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 27 35843886
2019 TRIM4 is associated with neural tube defects based on genome-wide DNA methylation analysis. Clinical epigenetics 17 30709423
2023 ABTB1 facilitates the replication of influenza A virus by counteracting TRIM4-mediated degradation of viral NP protein. Emerging microbes & infections 12 37823597
2020 TRIM4 interacts with TRPM8 and regulates its channel function through K423-mediated ubiquitination. Journal of cellular physiology 9 33037615
2024 TRIM4 enhances small-molecule-induced neddylated-degradation of CORO1A for triple negative breast cancer therapy. Theranostics 7 39629122
2022 TRIM4-mediated ubiquitination of NSP2 restricts porcine reproductive and respiratory syndrome virus proliferation. BMC veterinary research 5 35637527
2023 TRIM4 Expression Related to Malignant Progression and Cisplatin Resistance in Osteosarcoma. Applied biochemistry and biotechnology 2 37115387
2022 Sequence Analysis of Macaca mulatta TRIM4 and Its Role in the Interferon Pathway. Frontiers in veterinary science 2 35242838
2024 The TRIM4 E3 ubiquitin ligase degrades TPL2 and is modulated by oncogenic KRAS. Cell reports 1 39178114
2024 TRIM4 modulates the ubiquitin-mediated degradation of hnRNPDL and weakens sensitivity to CDK4/6 inhibitor in ovarian cancer. Frontiers of medicine 1 39643799
2026 The E3 ubiquitin ligase TRIM4 promotes the proliferation of glioblastoma by inhibiting ABTB1-mediated CDK1 ubiquitination. International journal of biological macromolecules 0 42034143

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