Affinage

RNF144B

E3 ubiquitin-protein ligase RNF144B · UniProt Q7Z419

Length
303 aa
Mass
33.7 kDa
Annotated
2026-06-10
17 papers in source corpus 13 papers cited in narrative 13 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RNF144B is a RING-IBR-RING (RBR) E3 ubiquitin ligase that operates downstream of the p53/p63 transcription factor network to control cell fate and innate immune signaling (PMID:12853982, PMID:38685100). It is a direct transcriptional target of p53 and of ΔNp63α, and its RING-IBR-RING domain engages the E2 conjugating enzymes UbcH7 and UbcH8 to drive substrate ubiquitination (PMID:12853982, PMID:16427630, PMID:23128396). As a multi-substrate ligase, RNF144B promotes degradation of the cyclin-dependent kinase inhibitor p21WAF1/CIP1 and of ΔNp63α itself, the latter forming an auto-regulatory feedback loop required for normal keratinocyte proliferation and differentiation (PMID:12853982, PMID:23128396). In apoptosis, RNF144B ubiquitinates Bax to restrain spontaneous cell death, and upon apoptotic stimulation it translocates from the cytosol to mitochondrial domains enriched in activated Bax (PMID:20300062). RNF144B is a recurrent negative regulator of innate immune signaling: it binds TBK1 through its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and downstream IRF3/IFN-β activation; it directs K27/K33-linked ubiquitination of MDA5 at K23/K43 within the CARDs to route MDA5 for p62-dependent autophagic degradation, limiting antiviral interferon responses; and it drives K48-linked ubiquitination and proteasomal degradation of TRAF3 to restrain NF-κB and MAPK signaling (PMID:31509299, PMID:39285245, PMID:41903812). Acting downstream of TP53, RNF144B functions as a tumor suppressor that maintains genomic stability, with its loss producing chromosomal instability and mitotic defects (PMID:38685100).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 2003 Medium

    Established RNF144B as a p53-inducible E3 ligase, linking it to the tumor suppressor transcriptional program and identifying p21WAF1 as a degradation substrate.

    Evidence In vitro E3 ligase assay, Co-IP of p53RFP-p21WAF1, p53 transcriptional reporter

    PMID:12853982

    Open questions at the time
    • E2 partner and ubiquitin linkage type not defined
    • in vivo relevance of p21 turnover untested
  2. 2006 Medium

    Mapped E2 enzyme specificity (UbcH7/UbcH8, not UbcH5) to the RING-IBR-RING domain and showed RNF144B-induced apoptosis is caspase-independent and separable from its ligase activity.

    Evidence Co-IP for E2 binding, domain deletion mutants, apoptosis assays in cell lines

    PMID:16427630

    Open questions at the time
    • substrate ubiquitinated during apoptosis not identified
    • mechanism of caspase-independent death by the C-terminal domain unresolved
  3. 2010 High

    Defined RNF144B as a regulator of Bax stability that protects against spontaneous apoptosis and relocalizes to mitochondria to engage activated Bax during cell death.

    Evidence Reciprocal Co-IP with activated Bax, Bax mutant epistasis in HCT116 Bax-null cells, ubiquitination assays, subcellular fractionation/live imaging

    PMID:20300062

    Open questions at the time
    • ubiquitin linkage type on Bax not defined
    • trigger for cytosol-to-mitochondria translocation not mechanistically resolved
  4. 2012 High

    Placed RNF144B in a ΔNp63α auto-regulatory feedback loop in keratinocytes, showing it degrades both p21WAF1 and ΔNp63α to enable proliferation and differentiation.

    Evidence ΔNp63α transcriptional reporter, reciprocal Co-IP, proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation readout

    PMID:23128396

    Open questions at the time
    • ubiquitin linkage on ΔNp63α not characterized
    • in vivo epidermal phenotype untested
  5. 2016 Medium

    Implicated RNF144B in inflammasome priming, showing it is LPS-inducible in human (not mouse) macrophages and selectively promotes IL-1β transcription.

    Evidence siRNA silencing in primary human macrophages, cytokine qPCR, CAGE promoter analysis, human/mouse comparison

    PMID:26819317

    Open questions at the time
    • molecular target mediating IL-1β induction not identified
    • ligase activity not connected to this phenotype
  6. 2018 Medium

    Showed RNF144B protein is stabilized by GSK3β-dependent phosphorylation, coupling its abundance to proliferative signaling in endometrial cancer.

    Evidence GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, proliferation assays

    PMID:29724995

    Open questions at the time
    • specific phosphorylation site not mapped
    • direct vs indirect role of GSK3β not established
  7. 2019 Medium

    Identified RNF144B as a negative regulator of TBK1-IRF3 signaling, binding TBK1 via its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and IFN-β production.

    Evidence Co-IP with domain mapping (IBR-SDD), siRNA knockdown, Western blot for phospho/ubiquitination status, IFN-β ELISA

    PMID:31509299

    Open questions at the time
    • whether ligase activity is required for TBK1 inhibition unclear
    • in vivo confirmation absent at this stage
  8. 2022 Medium

    Linked RNF144B to spermatogonial stem cell maintenance through an FCER2/NOTCH2/HES1 axis governing proliferation and survival.

    Evidence RNA-seq, Co-IP (RNF144B-FCER2, FCER2-N2ICD), siRNA knockdown, FCER2 overexpression rescue, proliferation/apoptosis assays

    PMID:35699595

    Open questions at the time
    • whether FCER2 is a ubiquitination substrate not tested
    • ligase-dependence of the NOTCH2 effect unknown
  9. 2023 Medium

    Provided in vivo evidence that RNF144B tunes macrophage immune signaling, with myeloid-specific loss impairing TBK1 activation while enhancing NF-κB and worsening sepsis outcomes.

    Evidence Myeloid conditional knockout mice, Co-IP (Rnf144b-TBK1), Western blot, cytokine measurement, in vivo sepsis model

    PMID:37088470

    Open questions at the time
    • molecular basis for divergent TBK1 vs NF-κB effects not resolved
    • substrate driving NF-κB suppression not identified at this point
  10. 2023 Medium

    Showed in ovarian cancer that HDAC3 induces RNF144B, which degrades the p21/p53 complex to promote a pro-tumorigenic phenotype, reinforcing p21 as a key substrate.

    Evidence Co-IP (RNF144B/p21/p53), cycloheximide chase, transcriptomics, knockdown/overexpression, in vivo tumor model

    PMID:37992458

    Open questions at the time
    • ubiquitin linkage on the p21/p53 complex not defined
    • apparent oncogenic role here contrasts with tumor-suppressor findings, context dependence unexplained
  11. 2024 High

    Defined the molecular mechanism of RNF144B antiviral restraint: K27/K33-linked ubiquitination of MDA5 at K23/K43 in the CARDs directs p62-mediated autophagic degradation, with knockout boosting interferon and antiviral protection.

    Evidence Co-IP, ubiquitination linkage/site mapping, Rnf144b knockout mice, EMCV challenge, IFN assays, autophagy analysis

    PMID:39285245

    Open questions at the time
    • E2 partner driving K27/K33 linkages not specified
    • interplay with the TBK1 branch of the same pathway not integrated
  12. 2024 Medium

    Established RNF144B as a TP53-downstream tumor suppressor maintaining genomic stability, with loss causing chromosomal instability and mitotic defects.

    Evidence Knockout in human/mouse non-transformed and cancer cells, proteomics/transcriptomics, cell cycle and DNA damage assays, in vivo tumor models

    PMID:38685100

    Open questions at the time
    • specific degradation substrates driving genomic stability not pinpointed
    • reconciliation with oncogenic context-dependent roles incomplete
  13. 2026 High

    Identified TRAF3 as a K48-linked ubiquitination substrate of RNF144B, showing degradation of TRAF3 restrains NF-κB/MAPK signaling and limits neuroinflammation after ischemic stroke.

    Evidence Co-IP, K48-linked ubiquitination assay, RNF144B knockout mice (MCAO), TRAF3 knockdown epistasis rescue, Western blot, immunofluorescence

    PMID:41903812

    Open questions at the time
    • E2 partner and structural basis of TRAF3 recognition not defined
    • relationship between TRAF3 and TBK1 branches of RNF144B immune regulation not integrated

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RNF144B's apparently opposing roles — tumor suppressor maintaining genomic stability versus context-dependent pro-proliferative ligase — are reconciled, and what determines its substrate selection across cell types, remains unresolved.
  • no unifying model for substrate choice across apoptotic, immune, and cell-cycle contexts
  • structural determinants of linkage-type specificity (K48 vs K27/K33) unknown
  • physiological hierarchy among Bax, p21, ΔNp63α, MDA5, TRAF3 substrates undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016874 ligase activity 4 GO:0140096 catalytic activity, acting on a protein 4
Localization
GO:0005739 mitochondrion 1 GO:0005829 cytosol 1
Pathway
R-HSA-168256 Immune System 5 R-HSA-392499 Metabolism of proteins 4 R-HSA-5357801 Programmed Cell Death 2 R-HSA-1640170 Cell Cycle 1
Partners
BAXCDKN1AFCER2IFIH1TBK1TP53TP63TRAF3

Evidence

Reading pass · 13 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2010 IBRDC2 (RNF144B) is an IBR-type RING-finger E3 ubiquitin ligase that regulates Bax stability through ubiquitination-dependent degradation, protecting cells from spontaneous apoptosis. Upon apoptosis induction, IBRDC2 translocates from cytosol to mitochondrial domains enriched with active Bax, physically interacts with activated Bax, and this translocation requires both mitochondrial localization and apoptotic activation of Bax, and depends on Bcl-xL expression levels. Co-immunoprecipitation (physical interaction with activated Bax), active Bax-specific antibody staining, Bax mutant analysis in HCT116 Bax(-/-) cells, ubiquitination assays with IBRDC2 overexpression/knockdown, subcellular fractionation/live imaging The EMBO journal High 20300062
2003 p53RFP (RNF144B) is a transcriptional target of p53 with E3 ubiquitin ligase activity that interacts with and ubiquitinates p21WAF1, promoting its degradation. Its expression is negatively correlated with p21WAF1 protein levels, suggesting it regulates p21WAF1 stability. E3 ubiquitin ligase activity assay, co-immunoprecipitation (p53RFP–p21WAF1 interaction), expression correlation analysis, transcriptional reporter assay for p53 induction Oncogene Medium 12853982
2006 p53RFP (RNF144B) induces p53-dependent but caspase-independent apoptosis. Its RING-IBR-RING domain mediates interaction with E2 ubiquitin-conjugating enzymes UbcH7 and UbcH8 but not UbcH5. The conserved C-terminal domain (not the E3 ligase domain) is required and sufficient for apoptosis induction, indicating apoptosis does not require E3 ubiquitin ligase activity. Co-immunoprecipitation (E2 enzyme binding), domain deletion mutant analysis, apoptosis assays (caspase-independent), overexpression in cell lines FEBS letters Medium 16427630
2012 PIR2/RNF144B is a direct transcriptional target of ΔNp63α in keratinocytes. It binds and mediates proteasomal degradation of both p21WAF1/CIP1 and ΔNp63α itself, forming an auto-regulatory feedback loop. Depletion of PIR2/RNF144B impairs keratinocyte proliferation and differentiation with accumulation of p21WAF1/CIP1. Transcriptional reporter assay (ΔNp63α target), co-immunoprecipitation (PIR2–ΔNp63α binding), proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation phenotypic readout Oncogene High 23128396
2019 RNF144B interacts with TBK1 through its IBR domain binding the scaffold/dimerization domain (SDD) of TBK1, inhibiting TBK1 phosphorylation and K63-linked polyubiquitination, which leads to TBK1 inactivation, IRF3 dephosphorylation, and reduced IFN-β production in response to LPS stimulation. RNF144B knockdown increases IRF3 activation and IFN-β production. Co-immunoprecipitation (RNF144B–TBK1 interaction), domain mapping (IBR domain–SDD interaction), siRNA knockdown, Western blot for phosphorylation and ubiquitination status, IFN-β ELISA Journal of leukocyte biology Medium 31509299
2016 RNF144B is necessary for priming of inflammasome responses in primary human macrophages; specifically, it promotes LPS-inducible IL-1β mRNA expression but does not regulate several other LPS-inducible cytokines (e.g., IL-10, IFN-γ) or inflammasome components (procaspase-1, pro-IL-18). RNF144B is LPS-inducible in human but not mouse macrophages due to differences in transcription factor binding sites in the promoter. Gene silencing (siRNA) in primary human macrophages, cytokine mRNA quantification (qPCR), cap analysis of gene expression (CAGE) for promoter activity, comparative analysis across human and mouse cell types Journal of leukocyte biology Medium 26819317
2018 PIR2/RNF144B protein is stabilized via phosphorylation downstream of GSK3β activity, and this stabilization is necessary for proliferation of endometrial cancer cells in the absence of oestrogenic growth stimuli. Inactivation of GSK3β leads to loss of PIR2/RNF144B protein and consequent inhibition of cell proliferation. GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, cell proliferation assays Cell death & disease Medium 29724995
2022 RNF144B interacts with FCER2 in human spermatogonial stem cells (SSCs), and through FCER2 activates the NOTCH2/HES1 pathway (FCER2 pulls down NOTCH2 intracellular domain N2ICD). RNF144B knockdown reduces NOTCH2, FCER2, HES1, and HEY1 levels, inhibits SSC proliferation, and promotes apoptosis; these effects are reversed by FCER2 overexpression. RNA sequencing (target identification), co-immunoprecipitation (RNF144B–FCER2 and FCER2–N2ICD interactions), siRNA knockdown, FCER2 overexpression rescue, proliferation and apoptosis assays Journal of cellular physiology Medium 35699595
2023 RNF144B interacts with TBK1; in sepsis model, Rnf144b deficiency results in impaired TBK1 activation but enhanced NF-κB activation in macrophages. Conditional knockout of Rnf144b in myeloid cells leads to increased inflammatory cytokines after LPS/CpG stimulation, higher mortality, and exacerbated cardiac dysfunction in septic mice. Conditional knockout mice (myeloid-specific), co-immunoprecipitation (Rnf144b–TBK1), Western blot for TBK1 and NF-κB activation, cytokine measurement, in vivo sepsis model ESC heart failure Medium 37088470
2023 HDAC3 promotes RNF144B expression in ovarian cancer cells. RNF144B interacts with p21 and regulates degradation of the p21/p53 complex. RNF144B-mediated p21 degradation (assessed by cycloheximide chase) promotes ovarian cancer cell proliferation, migration, and invasion while inhibiting apoptosis. Co-immunoprecipitation (RNF144B/p21/p53 interaction), cycloheximide chase (p21 stability), transcriptome profiling, siRNA knockdown and overexpression, in vivo tumor model Tissue & cell Medium 37992458
2024 RNF144B specifically interacts with MDA5 and promotes K27/K33-linked polyubiquitination of MDA5 at lysine residues K23 and K43 (within the CARDs domain), which promotes autophagic degradation of MDA5 by p62. Rnf144b knockout in mice greatly promotes IFN production, inhibits EMCV replication, and significantly increases overall survival upon EMCV infection. Co-immunoprecipitation (RNF144B–MDA5 interaction), ubiquitination assay with linkage type and site identification (K27/K33, K23/K43), Rnf144b knockout mice, viral challenge (EMCV), IFN production assay, autophagy pathway analysis EMBO reports High 39285245
2024 RNF144B acts as a tumor suppressor downstream of TP53 in lung adenocarcinoma, mediating protein degradation associated with cell cycle progression, DNA damage response, and genomic stability. RNF144B deficiency induces chromosomal instability, mitotic defects, and resistance to cell cycle inhibitors that induce chromosomal instability. RNF144B knockout in human and mouse cells (non-transformed and cancerous), proteomics and transcriptomics analysis, proliferation/transformation assays, cell cycle analysis, DNA damage response assays, in vivo tumor models Journal of experimental & clinical cancer research Medium 38685100
2026 RNF144B interacts with TRAF3 and promotes its K48-linked ubiquitination and proteasomal degradation. In the absence of RNF144B (KO mice), TRAF3 is stabilized, leading to enhanced NF-κB and MAPK signaling pathway activation and exacerbated neuroinflammation after ischemic stroke. TRAF3 knockdown in RNF144B-deficient mice partially reversed neurological dysfunction and neuroinflammation post-MCAO. Co-immunoprecipitation (RNF144B–TRAF3 interaction), ubiquitination assay (K48-linked), RNF144B knockout mice (MCAO model), TRAF3 knockdown epistasis rescue, Western blot for NF-κB and MAPK activation, immunofluorescence Biochemical pharmacology High 41903812

Source papers

Stage 0 corpus · 17 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2010 IBRDC2, an IBR-type E3 ubiquitin ligase, is a regulatory factor for Bax and apoptosis activation. The EMBO journal 74 20300062
2003 p53RFP, a p53-inducible RING-finger protein, regulates the stability of p21WAF1. Oncogene 44 12853982
2006 The p53-inducible E3 ubiquitin ligase p53RFP induces p53-dependent apoptosis. FEBS letters 32 16427630
2012 PIR2/Rnf144B regulates epithelial homeostasis by mediating degradation of p21WAF1 and p63. Oncogene 25 23128396
2020 LINC00662 triggers malignant progression of chordoma by the activation of RNF144B via targeting miR-16-5p. European review for medical and pharmacological sciences 24 32096180
2019 RNF144B inhibits LPS-induced inflammatory responses via binding TBK1. Journal of leukocyte biology 24 31509299
2016 The E3 ubiquitin ligase RNF144B is LPS-inducible in human, but not mouse, macrophages and promotes inducible IL-1β expression. Journal of leukocyte biology 17 26819317
2022 RNF144B stimulates the proliferation and inhibits the apoptosis of human spermatogonial stem cells via the FCER2/NOTCH2/HES1 pathway and its abnormality is associated with azoospermia. Journal of cellular physiology 16 35699595
2018 Pir2/Rnf144b is a potential endometrial cancer biomarker that promotes cell proliferation. Cell death & disease 15 29724995
2024 The TP53-activated E3 ligase RNF144B is a tumour suppressor that prevents genomic instability. Journal of experimental & clinical cancer research : CR 10 38685100
2024 RNF144B negatively regulates antiviral immunity by targeting MDA5 for autophagic degradation. EMBO reports 10 39285245
2023 Rnf144b alleviates the inflammatory responses and cardiac dysfunction in sepsis. ESC heart failure 10 37088470
2023 RNF144B-mediated p21 degradation regulated by HDAC3 contribute to enhancing ovarian cancer growth and metastasis. Tissue & cell 7 37992458
2025 RNF144A and RNF144B: Important molecules for health. Open life sciences 2 40756570
2024 Integrated analysis ceRNA network of autophagy-related gene RNF144B in steroid-induced necrosis of the femoral head. Scientific reports 1 39567703
2026 RNF144B deficiency aggravated neuroinflammatory response induced by cerebral ischemic stroke via regulating TRAF3 ubiquitination. Biochemical pharmacology 0 41903812
2025 Characterization of RNF144B and PPP2R2A identified by a novel approach using TCGA data in ovarian cancer. Scientific reports 0 39948107

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