Affinage

RIPK3

Receptor-interacting serine/threonine-protein kinase 3 · UniProt Q9Y572

Length
518 aa
Mass
56.9 kDa
Annotated
2026-04-28
100 papers in source corpus 44 papers cited in narrative 44 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RIPK3 is a serine/threonine kinase that serves as a central signaling hub governing necroptosis, apoptosis, inflammation, and ferroptosis, with the outcome determined by its activation state, oligomeric assembly, and post-translational modifications. Upon RHIM-domain-dependent interaction with RIPK1, TRIF, or ZBP1, RIPK3 undergoes amyloid-like oligomerization and autophosphorylation to form the necrosome, wherein it phosphorylates MLKL at the C-lobe interface to execute necroptosis; phosphorylation at S165/T166 instead inactivates kinase activity and redirects RIPK3 to recruit RIPK1, FADD, and caspase-8 for apoptosis, while RIPK3 stability is regulated by PELI1- and TRIM25-mediated K48-linked ubiquitylation, PPM1B dephosphorylation at T231/S232, and PRMT1-mediated methylation at R486 (PMID:34811356, PMID:35256774, PMID:34029184, PMID:25751141, PMID:29883609, PMID:37005412). Beyond cell death, RIPK3 promotes NF-κB-dependent and NLRP3 inflammasome-driven cytokine production independently of MLKL, phosphorylates CaMKII to open the mitochondrial permeability transition pore, and phosphorylates FSP1 to inhibit its ferroptosis-suppressive activity downstream of ZBP1 sensing of mitochondrial Z-DNA (PMID:25367573, PMID:25693118, PMID:26726877, PMID:38493248). OASL scaffolds RIPK3–ZBP1 assembly via liquid-like phase condensation to facilitate amyloid fibril formation and efficient autophosphorylation during viral infection (PMID:36604592).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1999 High

    The initial identification of RIPK3 as a kinase that binds RIPK1 through its C-terminal domain and modulates TNF receptor signaling established it as a novel component of the TNFR1 complex with roles in NF-κB attenuation and caspase-dependent apoptosis.

    Evidence Co-immunoprecipitation, overexpression/reporter assays, and binding domain mapping in mammalian cells

    PMID:10358032

    Open questions at the time
    • No endogenous loss-of-function evidence
    • Kinase substrates unknown
    • Role in non-apoptotic death not yet recognized
  2. 2010 High

    Discovery that RIPK3 is the essential kinase for programmed necrosis (necroptosis) via RHIM-dependent necrosome formation with RIPK1, regulated by caspases and ubiquitination, and targeted by viral inhibitors, reframed RIPK3 from an NF-κB modulator to the central executioner of a distinct cell death pathway.

    Evidence RIP3-deficient mice, viral infection (MCMV with vIRA/M45), biochemical fractionation, enzymatic and ROS assays

    PMID:20354226 PMID:20413098

    Open questions at the time
    • Downstream substrate executing membrane rupture not yet identified
    • Whether RIPK3 functions independently of RIPK1 unknown
  3. 2013 High

    Demonstration that TLR3/TLR4-TRIF and DAI(ZBP1) activate RIPK3 through RHIM independently of RIPK1, and that phosphorylation at S204 drives RIP1-independent necroptosis, established multiple upstream nucleation pathways and key regulatory phosphosites.

    Evidence RHIM mutant analysis, RIP3 kinase inhibitors, RIPK1/RIPK3/MLKL KO cells, S204D phosphomimetic mutagenesis

    PMID:23913919 PMID:24019532 PMID:24059293

    Open questions at the time
    • MLKL phosphorylation mechanism by RIPK3 not structurally resolved
    • Regulation of S204 phosphorylation by upstream kinases/phosphatases unclear
  4. 2014 High

    A series of studies revealed that RIPK3 controls both necroptosis and apoptosis through distinct mechanisms: enforced oligomerization (not mere dimerization) activates MLKL-dependent necroptosis, while kinase-dead or inhibited RIPK3 recruits FADD-caspase-8 for apoptosis, and RIPK1 acts as both a positive seed and negative brake on spontaneous RIPK3 activation.

    Evidence Chemically induced oligomerization, kinase-dead D161N knock-in mice, small-molecule inhibitors, FADD/caspase-8/MLKL KO cells, RIPK1-KO epistasis

    PMID:24813849 PMID:24902899 PMID:24902904 PMID:25459880

    Open questions at the time
    • Structural basis of oligomer-to-MLKL recruitment not resolved
    • How kinase-dead RIPK3 conformationally exposes RHIM for apoptosis scaffold unclear
  5. 2014 High

    RIPK3 was shown to drive NLRP3 inflammasome activation and NF-κB-dependent cytokine production (RelB/p50, IL-1β) independently of MLKL and necroptotic death, establishing a non-lethal inflammatory signaling function.

    Evidence IAP-KO, MLKL-KO, and kinase-dead mutant macrophages and dendritic cells, inflammasome assays, in vivo arthritis and colitis models

    PMID:25367573 PMID:25693118

    Open questions at the time
    • Precise mechanism linking RIPK3 scaffolding to NLRP3 assembly not defined
    • Whether kinase activity contributes to inflammation in all contexts unresolved
  6. 2015 High

    Identification of PPM1B as the first RIPK3 phosphatase, dephosphorylating T231/S232 to prevent MLKL recruitment, demonstrated that necrosome signaling is reversibly controlled by a phosphatase checkpoint.

    Evidence Co-IP, in vitro phosphatase assays, site-directed mutagenesis, Ppm1b-KO mice with enhanced TNF-induced death

    PMID:25751141

    Open questions at the time
    • Whether additional phosphatases act on other RIPK3 phosphosites unknown
    • Tissue-specific relevance of PPM1B regulation not broadly characterized
  7. 2016 High

    RIPK3 was found to phosphorylate CaMKII as an alternative substrate to trigger mitochondrial permeability transition pore opening, defining a non-MLKL necroptotic pathway operative in the heart.

    Evidence In vitro kinase assay with CaMKII as substrate, RIP3-KO mice in cardiac ischemia-reperfusion, mPTP assays

    PMID:26726877

    Open questions at the time
    • Whether CaMKII phosphorylation by RIPK3 occurs outside cardiomyocytes not established
    • Stoichiometric relationship between MLKL- and CaMKII-dependent pathways not quantified
  8. 2018 High

    Multiple layers of RIPK3 protein turnover control were uncovered: PELI1 ubiquitylates active (T182-phosphorylated) RIPK3 at K363 for proteasomal degradation, and TRIM25 ubiquitylates RIPK3 at K501 via K48-linked chains, while the kinase domain was shown to homodimerize in a RAF-like manner to stimulate allosteric cis-autophosphorylation.

    Evidence E3 ligase–substrate Co-IP, mutagenesis of K363/T182 and K501, ubiquitin chain analysis, structural analysis of kinase domain dimer interface

    PMID:29883609 PMID:30131368 PMID:33953350

    Open questions at the time
    • Whether PELI1 and TRIM25 act redundantly or in distinct contexts unclear
    • Full ubiquitylation landscape of RIPK3 not mapped
  9. 2019 High

    RIPK3 was shown to restrict West Nile virus independently of necroptosis by promoting neuronal chemokine expression, and to act as a scaffold for ZBP1-driven NF-κB inflammatory signaling through K63/M1 ubiquitin chains on RIPK1, further separating its death-independent functions.

    Evidence Ripk3-KO vs. Mlkl-KO mice in WNV encephalitis, ZBP1 Co-IP with ubiquitin chain analysis, kinase inhibitor dissection

    PMID:28366204 PMID:36268590

    Open questions at the time
    • RIPK3 scaffold function specificity across different viral infections not systematically tested
    • Whether kinase activity contributes to antiviral chemokine induction unresolved
  10. 2021 High

    Crystal structures of the human RIPK3 kinase domain alone and in complex with MLKL pseudokinase revealed the C-lobe interface critical for complex assembly and species specificity, providing the first atomic-resolution view of necrosome-to-effector signaling, while phosphorylated MLKL was shown to disengage from RIPK3 prior to executing death.

    Evidence X-ray crystallography, structure-guided mutagenesis, monobody binding proteins, live-cell imaging

    PMID:33850121 PMID:34811356

    Open questions at the time
    • Full-length necrosome structure (RIPK1–RIPK3 amyloid core + MLKL) not determined
    • Dynamics of MLKL disengagement in different cell types not characterized
  11. 2021 High

    Phosphorylation of RIPK3 at S165/T166 was shown to be a molecular switch: it inactivates kinase activity while creating a platform for RIPK1-FADD-caspase-8 recruitment and apoptosis, with phospho-mimetic knock-in mice dying neonatally from spontaneous apoptosis.

    Evidence Phospho-resistant and phospho-mimetic knock-in mice, Co-IP, caspase activation assays, Hsp90/CDC37 manipulation

    PMID:34029184

    Open questions at the time
    • Kinase(s) responsible for S165/T166 phosphorylation in physiological contexts not identified
    • Structural basis of the necroptosis-to-apoptosis conformational switch not resolved
  12. 2022 High

    Super-resolution imaging revealed that necrosomes are mosaic amyloid-like fibrils of RIPK1 and RIPK3 oligomers, with RIPK3 tetramers or larger being the minimal units for MLKL recruitment, and RIPK1 autophosphorylation controlling ordered oligomer growth.

    Evidence Super-resolution microscopy, oligomerization assays, autophosphorylation mutants, functional cell death assays

    PMID:35256774

    Open questions at the time
    • Whether oligomer stoichiometry varies between RHIM nucleators (RIPK1 vs. ZBP1 vs. TRIF) not tested
    • In vivo oligomer architecture not examined
  13. 2023 High

    OASL was identified as a scaffold that nucleates RIPK3–ZBP1 assembly through liquid-like phase condensation, promoting amyloid fibril formation and RIPK3 autophosphorylation, and PRMT1 methylation of RIPK3 at R486 was found to inhibit RIPK1–RIPK3 necrosome assembly.

    Evidence Phase condensation and amyloid fibril assays, Oasl1-KO mice, in vitro methylation assay with PRMT1, R486 mutagenesis, site-specific antibody

    PMID:36604592 PMID:37005412

    Open questions at the time
    • Whether OASL also scaffolds RIPK1-seeded necrosomes unknown
    • Physiological conditions regulating PRMT1-mediated RIPK3 methylation not defined
  14. 2024 High

    RIPK3 was found to phosphorylate FSP1, inhibiting its anti-ferroptotic enzymatic activity downstream of ZBP1 sensing of mitochondrial Z-DNA, establishing RIPK3 as a dual-death kinase that can independently execute necroptosis (via MLKL) and ferroptosis (via FSP1) in the same cell.

    Evidence Endothelial-specific Zbp1-KO and Ripk3-KO mice, MLKL-KO mice, FSP1 phosphorylation assays, Z-DNA detection

    PMID:38493248

    Open questions at the time
    • FSP1 phosphorylation site(s) by RIPK3 not mapped at residue level
    • Whether FSP1 phosphorylation by RIPK3 occurs outside the ZBP1 context unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • Major unresolved questions include the full-length necrosome structure at atomic resolution, how the same RIPK3 molecule selectively switches among necroptosis, apoptosis, ferroptosis, and non-lethal inflammatory signaling, and whether therapeutic RIPK3 inhibition can be achieved without triggering compensatory apoptosis.
  • No cryo-EM or crystallographic structure of the intact RIPK1-RIPK3 amyloid necrosome
  • Cell-type-specific hierarchy of RIPK3 substrates (MLKL vs. CaMKII vs. FSP1) not systematically mapped
  • Safe therapeutic window for RIPK3 kinase inhibitors remains undefined due to apoptosis-switching risk

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 8 GO:0098772 molecular function regulator activity 4
Localization
GO:0005829 cytosol 3 GO:0005634 nucleus 2 GO:0005739 mitochondrion 2
Pathway
R-HSA-5357801 Programmed Cell Death 11 R-HSA-168256 Immune System 7 R-HSA-162582 Signal Transduction 5
Partners
FADDMLKLPELI1PPM1BRIPK1TRIFTRIM25ZBP1
Complex memberships
Necrosome (RIPK1-RIPK3 amyloid-like complex)RIPK3-MLKL complexTRIF-RIPK3 complexZBP1-RIPK3 complex

Evidence

Reading pass · 44 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 RIP3 was identified as a novel kinase containing an N-terminal kinase domain; it binds RIP1 through its unique C-terminal segment, is recruited to the TNF receptor-1 signaling complex via this interaction, attenuates RIP1- and TNFR1-induced NF-κB activation, and induces apoptosis when overexpressed, selectively binding large prodomain initiator caspases. Co-immunoprecipitation, overexpression/reporter assays, binding domain mapping The Journal of biological chemistry High 10358032
2010 Murine cytomegalovirus infection induces RIP3-dependent necrosis through RHIM-dependent interactions; viral vIRA (M45) targets RIP3 and disrupts RIP3-RIP1 interactions, suppressing both TNF-dependent necroptosis and virus-induced necrosis. RIP3 kinase activity and RHIM interactions are required for virus-associated necrosis, which proceeds independently of RIP1. Genetic knockout (RIP3-deficient mice), viral infection assays, co-immunoprecipitation, in vivo attenuation studies Cell host & microbe High 20413098
2010 RIP1 and RIP3 form a necrosome complex upon TNF stimulation that drives programmed necrosis; RIP3 activates enzymes controlling glycolytic flux and glutaminolysis, and the necrosome promotes mitochondrial complex I-mediated ROS production. RIP3 activity is regulated by caspases and ubiquitination. Biochemical fractionation, enzymatic assays, caspase/ubiquitination studies, ROS measurement Science signaling High 20354226
2011 RIP3 mediates programmed necrosis of intestinal epithelial cells (IECs); genetic deletion of RIP3 prevents spontaneous epithelial cell necrosis, Paneth cell loss, and colitis in FADD-knockout mice. CYLD deubiquitinase regulates cellular necrosis in this context and is upstream of RIP3-dependent death. Genetic epistasis (FADD IEC-KO × RIP3-KO double mutant mice), histology, immunofluorescence Nature High 21804564
2013 TLR3 and TLR4 directly activate programmed necrosis through a RHIM-dependent association of the TIR adaptor TRIF with RIP3 kinase; in fibroblasts, this TRIF-RIP3-MLKL pathway proceeds independently of RIP1 or its kinase activity but requires MLKL downstream of RIP3. Small molecule RIP3 kinase inhibitors block necroptosis induced by RIP1-RIP3, DAI-RIP3, and TRIF-RIP3 complexes. RHIM mutant analysis, RIP3 kinase inhibitors, RIP1/RIP3/MLKL knockout cells, Co-IP The Journal of biological chemistry High 24019532
2013 RIP3 activation is regulated by phosphorylation, ubiquitination, and caspase-mediated cleavage; these post-translational modifications coordinately regulate assembly of the necrosome, a macromolecular signaling complex. RIP3 can also promote inflammation independently of its pro-necrotic activity. Biochemical analysis, mutagenesis, pathway dissection in KO cells Genes & development High 23913919
2013 Phosphorylation of RIP3 at S204 generates a phosphomimetic form (S204D) that drives programmed necrosis independently of RIP1 and is refractory to necrostatin-1. Ser89 of RIP1 is an inhibitory phosphoacceptor site whose alanine substitution enhances RIP1 kinase activity and TNF-induced programmed necrosis without affecting RIP1-RIP3 necrosome formation. Site-directed mutagenesis, kinase activity assays, siRNA knockdown, cell death assays The Biochemical journal High 24059293
2014 RIP3 kinase inhibitors unexpectedly induce apoptosis via RHIM-driven recruitment of RIP1 to assemble a Casp8-FADD-cFLIP complex, independently of pronecrotic kinase activities and MLKL. The kinase-dead D161N RIP3 mutant spontaneously induces apoptosis, causing perinatal lethality in homozygous D161N mice, whereas K51A mutant mice are viable—demonstrating that RIP3 holds necroptosis and apoptosis in balance through a Ripoptosome-like platform. Small-molecule inhibitors, kinase-dead knock-in mice, Co-IP, FADD/caspase-8 genetic analysis Molecular cell High 25459880
2014 RIPK1 deletion leads to systemic inflammation due to unrestrained RIPK3-MLKL-dependent necroptosis; deletion of RIPK3 or MLKL (but not caspase-8) prevents extracellular release of the necroptotic DAMP IL-33 and reduces MyD88-dependent inflammation. RIPK1 thus negatively regulates RIPK3 to limit inflammation. Genetic epistasis with Ripk1/Ripk3/Mlkl/Casp8 knockout mice, cytokine measurements, bone marrow transplant Cell High 24813849
2014 RIPK1 positively seeds RHIM-dependent RIPK3 oligomers upon receptor signaling but intrinsically suppresses spontaneous RIPK3 activation in the cytosol by controlling RIPK3 oligomerization. Chemically enforced oligomerization of RIPK3 is sufficient to induce necroptosis independently of RHIM domain, TNF, or RIPK1. RIPK3 dimer formation alone is insufficient to trigger death. Chemically induced dimerization/oligomerization system, RIPK1-KO cells, RHIM mutants, live-cell imaging Cell death and differentiation High 24902904
2014 RIPK3 overexpression in cardiomyocytes is sufficient to induce the formation of a RIP1-RIP3 complex and trigger necroptosis; in vivo, RIP3-deficient mice show better cardiac function and less adverse remodeling after myocardial infarction. Adenoviral overexpression in neonatal rat cardiomyocytes, Co-IP, RIP3-/- mouse model of myocardial infarction, MRI Cardiovascular research Medium 24920296
2014 When RIPK3 is dimerized using a coumermycin system, it induces cell death by an MLKL-dependent mechanism and also by FADD/caspase-8/RIPK1-dependent mechanism; catalytically active RIPK3 kinase domains are required for MLKL-dependent but not caspase-8-dependent death. The mode of death is determined by the availability of FADD, caspase-8, and MLKL. Chemically induced dimerization, FADD/caspase-8/MLKL KO cells, kinase-dead mutants Cell death and differentiation High 24902899
2014 RIPK3 promotes NLRP3 inflammasome and IL-1β inflammatory responses independently of MLKL and necroptotic cell death; in the absence of IAPs, LPS triggers RIPK3 to activate caspase-8 promoting apoptosis and NLRP3-caspase-1 activation independently of RIPK3 kinase activity and MLKL. IAPs and TRIF regulate RIPK3 and MLKL ubiquitylation. IAP-KO and MLKL-KO cells, kinase-dead mutants, caspase-8 inhibition, NLRP3 inflammasome assays, arthritis mouse model Nature communications High 25693118
2014 RIPK3 controls NF-κB subunit RelB and p50 activation and caspase-1-mediated IL-1β processing in dendritic cells independently of its pronecrotic activity, thereby regulating inflammatory cytokine expression and injury-induced tissue repair. Ripk3-/- bone marrow-derived dendritic cells, LPS stimulation, NF-κB reporter assays, adoptive DC transfer, DSS colitis model Immunity High 25367573
2015 Ppm1b (protein phosphatase 1B) is a phosphatase for Rip3 that dephosphorylates Rip3 at Thr231/Ser232 in the necrosome, preventing recruitment of MLKL and thereby suppressing necroptosis. Ppm1b-deficient mice show enhanced TNF-induced death with elevated Rip3 phosphorylation in a Rip3-dependent manner. Co-IP, phosphatase assays, site-directed mutagenesis of phosphorylation sites, Ppm1b-KO mice Nature cell biology High 25751141
2016 RIP3 phosphorylates CaMKII (Ca2+-calmodulin-dependent protein kinase II) as a substrate—via phosphorylation or oxidation—to trigger opening of the mitochondrial permeability transition pore (mPTP) and myocardial necroptosis, defining an RIP3-CaMKII-mPTP pathway distinct from the canonical RIP1/MLKL pathway. In vitro kinase assay, RIP3-/- and CaMKII inhibitor mouse models, mPTP assays, cardiac ischemia-reperfusion model Nature medicine High 26726877
2016 Crystal cytotoxicity (calcium oxalate, monosodium urate, etc.) requires RIPK3-MLKL-mediated necroptosis; RNAi knockdown of RIPK3 or MLKL blocks crystal-induced cell death, and RIPK3- or MLKL-deficient mice are protected from oxalate crystal-induced acute kidney injury driven by TNF-α. RNAi knockdown, RIPK3-KO and MLKL-KO mice, cell death assays, pharmacological inhibitors Nature communications High 26817517
2016 Proteasome inhibitors activate the RIPK3-MLKL necroptotic pathway without requiring caspase inhibition, through a mechanism requiring an intact RHIM domain. Ubiquitin-proteasome system regulates RIPK3 stability through K48-linked ubiquitination at Lys264; when MLKL recruitment to RIPK3 is restricted, proteasome inhibitors induce RIPK3-dependent apoptosis. Proteasome inhibitor treatment, RHIM mutants, K264R RIPK3 mutant, MLKL restriction, cell death assays The Journal of biological chemistry Medium 26786097
2016 RIPK3 deficiency in adipose tissue promotes increased caspase-8-dependent adipocyte apoptosis and WAT inflammation, impairing insulin signaling and causing glucose intolerance, demonstrating that RIPK3 maintains adipose tissue homeostasis by suppressing caspase-8-dependent apoptosis. Ripk3-/- mice on high-fat diet, caspase-8 activity assays, insulin signaling analysis, metabolic phenotyping Nature communications Medium 27323669
2017 RIPK3 restricts West Nile virus pathogenesis and promotes neuronal chemokine expression and CNS immune cell recruitment independently of necroptotic cell death, as MLKL-KO mice are unaffected while Ripk3-/- mice show enhanced mortality with suppressed neuroinflammation. Ripk3-/-, Mlkl-/-, and Mlkl-/-Casp8-/- mouse models of WNV encephalitis, chemokine measurement, immune cell phenotyping Cell High 28366204
2017 RIPK3 mediates two distinct processes in FADD-deficient mice: RIPK3-MLKL-driven necroptosis during embryogenesis (rescued by kinase-dead Ripk3Δ mutation) and RIPK3-mediated postnatal inflammation causing lethality within 1 day of birth. Ripk3Δ/Δ kinase-dead knock-in mice crossed to Fadd-/- mice, phenotypic analysis of embryonic and postnatal survival Cell reports High 28445730
2018 PELI1 E3 ubiquitin ligase targets kinase-active RIP3 for K48-linked polyubiquitylation at K363 and proteasomal degradation; this requires phosphorylation of RIP3 at T182, which allows interaction with the PELI1 FHA domain. PELI1-mediated RIP3 degradation prevents cell death from RIP3 hyperactivation. Co-IP, mutagenesis (T182, K363), ubiquitin chain analysis, PELI1 overexpression/KD, proteasome inhibitor experiments Molecular cell High 29883609
2018 The RIPK3 kinase domain homodimerizes through a surface structurally similar to RAF family members; kinase domain dimerization stimulates RIPK3 activation via cis-autophosphorylation in a noncatalytic, allosteric manner. Mutation of the dimer interface impairs dimerization and necroptosis. The D161N mutant enhances this dimerization-dependent noncatalytic activity. Structural analysis, dimerization interface mutants, autophosphorylation assays, RIPK1-KO mice, RIPK3 D161N heterozygous/homozygous studies Science signaling High 30131368
2018 TRIM25 E3 ubiquitin ligase interacts with RIP3 via its SPRY domain and mediates K48-linked polyubiquitination of RIP3 at K501 through its RING domain, promoting proteasomal degradation of RIP3 and thereby suppressing TNF-induced cell necrosis. Co-IP, domain deletion mutants (SPRY, RING), ubiquitination assays, K501 mutagenesis, TRIM25 KO cells Cell death and differentiation High 33953350
2018 RIP3 nuclear translocation and co-localization with AIF after global cerebral ischemia/reperfusion injury leads to formation of an RIP3-AIF complex whose nuclear translocation causes ischemic neuronal DNA degradation; this process is blocked by necrostatin-1 and the autophagy inhibitor 3-MA but not by caspase-3 inhibitor. Co-immunoprecipitation, immunofluorescence, necrostatin-1 and autophagy inhibitor pharmacology, TUNEL assay in rat I/R model Scientific reports Medium 27377128
2018 Nuclear RIPK3 and MLKL continuously shuttle between the nucleus and cytoplasm. During TNF-induced necroptosis, nuclear MLKL becomes phosphorylated and oligomerized; inhibition of nuclear export retains RIPK3/MLKL in the nucleus, prevents cytosolic RIPK3/MLKL oligomerization, and reduces necroptotic cell death—indicating that nuclear passage regulates cytosolic necrosome formation. Nuclear export inhibitors, subcellular fractionation, immunofluorescence, phospho-MLKL detection, cell death assays Communications biology Medium 30271893
2019 CK1γ1 and CK1γ3 are recruited to the necrosome containing RIPK1, RIPK3, and MLKL; in vitro assays show CK1γ directly phosphorylates RIPK3 to affect its activity. Autophosphorylated CK1γ3 at Ser344/345 is detected in the necrosome and is required for necroptosis. Kinase screen (546 kinases), Co-IP, in vitro phosphorylation assay with purified proteins, CK1γ inhibitor in vivo model Cell death & disease Medium 31801942
2019 ZBP1 (DAI) associates with RIPK1 and RIPK3 during viral infection; RIPK3 also acts as a scaffold in ZBP1-induced inflammatory signaling via K63- and M1-linked ubiquitin chains on RIPK1 and ZBP1, promoting TAK1- and IKK-mediated cytokine production independently of cell death. Co-IP, ubiquitin chain analysis, RIPK1/RIPK3 kinase inhibitors, ZBP1-overexpressing HT29 cells EMBO reports Medium 36268590
2019 RIPK3-MLKL pathway activation suppresses intracellular Listeria replication in a cell-death-independent manner; MLKL directly binds to Listeria and inhibits replication in the cytosol. Ripk3-/- mice show dramatically increased systemic Listeria infection. Ripk3-/- mice, intracellular replication assays, MLKL-Listeria direct binding assay, pMLKL detection without cell death The Journal of cell biology High 30975711
2020 RIPK3 deficiency in TAMs reduces ROS, inhibits caspase-1-mediated cleavage of PPARδ, enabling PPARδ activation, which facilitates fatty acid oxidation and M2 polarization in the tumor microenvironment. RIPK3-KO macrophages, caspase-1 cleavage assay, PPARδ activation assays, FAO measurement, tumor model Cancer immunology research Medium 32122992
2020 ER stress activates RIPK3 in β-cells leading to NF-κB-mediated proinflammatory gene expression (IL-1β); this RIPK3-mediated IL-1β induction occurs independently of necroptosis and is conserved across zebrafish, mouse, and human islet models. RIPK3 inhibitor treatment, ripk3 KO in zebrafish model, mouse islet ex vivo, human islet xenograft, NF-κB reporter assays Science advances Medium 33355143
2020 BPA induces endothelial cell necroptosis via an RIP3/CaMKII-dependent pathway: silencing RIP3 reverses BPA-induced necroptosis and CaMKII activation, while inhibiting CaMKII (KN-93) does not affect RIP3 expression but decreases necroptosis, placing RIP3 upstream of CaMKII. siRNA RIP3 silencing, CaMKII inhibitor KN-93, cell death assays, in vivo BPA mouse model Scientific reports Medium 32144343
2021 The crystal structure of human RIPK3 kinase domain alone and in complex with the MLKL pseudokinase was determined; the structure reveals how human RIPK3 maintains MLKL in an inactive conformation prior to necroptosis induction, and residues at the RIPK3:MLKL C-lobe interface are crucial for complex assembly and necroptotic signaling, explaining strict species specificity. X-ray crystallography, structure-guided mutagenesis, cell death functional assays in human cells Nature communications High 34811356
2021 Phosphorylated MLKL undergoes conformational change and disengages from RIPK3 prior to cell death; two distinct conformations of the MLKL pseudokinase domain were identified by crystal structures, and MLKL disengagement from RIPK3 was confirmed using monobody-27 that binds an epitope overlapping the RIPK3 binding site only after phosphorylated MLKL disengages. Monobody binding proteins, crystal structures of MLKL pseudokinase domain, live-cell imaging, functional necroptosis assays Nature communications High 33850121
2021 RIP3 translocates into mitochondria in response to renal ischemia-reperfusion, interacts with inner mitochondrial membrane protein Mitofilin, and promotes its degradation, resulting in mtDNA release that activates the cGAS-STING-p65 pathway and promotes transcription of pro-inflammatory markers. Mitochondrial fractionation, Co-IP of RIP3-Mitofilin, RIP3-/- mice, cGAS-STING pathway analysis, RIP3 overexpression in HK-2 cells Cells Medium 35741025
2021 RIPK3 kinase phosphorylates serine 165/threonine 166 sites on its own kinase activation loop; when these sites are phosphorylated, RIPK3 kinase activity is inactivated but RIPK3 gains the ability to recruit RIPK1, FADD, and caspase-8 to form an apoptosis-inducing complex. This pathway mediates PGF2α-induced luteal regression. Phospho-resistant S165A/T166A mice fail to respond to PGF2α, while phospho-mimicking S165D/T166E mice undergo spontaneous apoptosis and die shortly after birth. Phospho-resistant and phospho-mimicking knock-in mice, Co-IP, caspase activation assays, Hsp90/CDC37 manipulation eLife High 34029184
2022 RIP1 and RIP3 form fibrous amyloid-like oligomers in necrosomes; super-resolution microscopy reveals that cellular necrosomes consist of mosaic complexes of RIP1 and RIP3 oligomers, with rod-shaped large mosaics required for functional necroptosis or apoptosis. RIP3 oligomers of tetramer size or above are the domains that allow MLKL recruitment and oligomerization. RIP1 autophosphorylation controls ordered oligomerization and is required for RIP1-initiated RIP3 homo-oligomerization. Super-resolution microscopy, oligomerization assays, autophosphorylation mutants, functional cell death assays Nature cell biology High 35256774
2022 RIPK3 inhibits autophagic flux during STING activation; RIPK3 knockout enhances STING autophagy and thereby suppresses STING signaling. Separately, MLKL regulates STING bidirectionally—when MLKL pro-necroptotic activity is blocked, MLKL bound to activated STING is secreted extracellularly, restricting TBK1 and IRF3 recruitment. RIPK3-KO HT-29 cells, autophagy flux assays, STING pathway readouts, MLKL secretion analysis Clinical and translational medicine Medium 37475188
2022 RIPK3-MLKL signaling in CKD promotes TGFβ-induced translocation of RIPK3 and MLKL to mitochondria, causing mitochondrial dysfunction and ROS production that activates CaMKII, which phosphorylates Smad2/3 to drive pro-fibrotic gene expression (αSMA, collagen). Ripk3-/- and Mlkl-/- mice, mitochondrial fractionation, CaMKII inhibition, Smad2/3 phosphorylation assays, kidney fibrosis models Matrix biology Medium 35964866
2022 RIPK3 inactivation increases susceptibility to 15LOX/PEBP1-driven ferroptosis; PEBP1 forms a complex with RIP3 and inhibits necroptosis. When 15LOX expression is elevated, it outcompetes RIP3 for PEBP1 binding, diverting PEBP1 from RIP3 and promoting PUFA-PE oxidation and ferroptosis. Rip3K51A kinase-inactive knock-in mice, Co-IP of PEBP1-RIP3 and PEBP1-15LOX, redox lipidomics, computational structural analysis Redox biology Medium 35101798
2023 OASL protein scaffolds RIPK3-ZBP1 assembly via liquid-like phase condensation to facilitate amyloid-like fibril formation and autophosphorylation of RIPK3, thereby promoting MLKL phosphorylation and necroptosis during viral infection. Mice deficient in Oasl1 have severely impaired RIPK3-ZBP1-driven necroptosis. Phase condensation assays, amyloid fibril detection, RIPK3 autophosphorylation assays, Oasl1-KO mice, viral infection models Nature cell biology High 36604592
2023 PRMT1 methyltransferase methylates RIP3 at R486 (human) / R479 (mouse); this methylation inhibits the interaction of RIP3 with RIP1 and suppresses RIP1-RIP3 necrosome complex formation, blocking RIP3 phosphorylation and necroptosis activation. In vitro methylation assay, Co-IP, phosphorylation assays, methylation-deficiency RIP3 mutant, site-specific antibody (RIP3ADMA) Cell death & disease High 37005412
2024 RIPK3 phosphorylates FSP1 to inhibit its enzymatic activity, promoting ferroptosis downstream of ZBP1 sensing of mitochondrial Z-form DNA; in endothelial cells, ZBP1 interacts with RIPK3, and RIPK3 has dual roles—phosphorylating MLKL to induce necroptosis and phosphorylating FSP1 to promote ferroptosis. Zbp1-KO and Ripk3-KO endothelial-specific mice, MLKL-KO mice, FSP1 phosphorylation assays, Z-DNA detection Cell death and differentiation High 38493248
2024 RIPK3 induces diabetic podocytopathy by regulating mitochondrial fission via the PGAM5-Drp1 pathway through MLKL; RIPK3 inhibition attenuates mitochondrial fission and dysfunction by decreasing pMLKL, PGAM5, and p-Drp1 S616 and mitochondrial Drp1 translocation. Ripk3-KO mice on high-fat diet, RIPK3 inhibition, MLKL phosphorylation assay, PGAM5 and Drp1 phosphorylation analysis, single-cell RNA sequencing, cultured podocytes Metabolism: clinical and experimental Medium 39089491

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2013 Toll-like receptor 3-mediated necrosis via TRIF, RIP3, and MLKL. The Journal of biological chemistry 802 24019532
2016 CaMKII is a RIP3 substrate mediating ischemia- and oxidative stress-induced myocardial necroptosis. Nature medicine 664 26726877
2015 RIPK3 promotes cell death and NLRP3 inflammasome activation in the absence of MLKL. Nature communications 566 25693118
2014 RIP3 induces apoptosis independent of pronecrotic kinase activity. Molecular cell 530 25459880
2014 RIPK1 regulates RIPK3-MLKL-driven systemic inflammation and emergency hematopoiesis. Cell 514 24813849
2011 FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation. Nature 511 21804564
2010 Virus inhibition of RIP3-dependent necrosis. Cell host & microbe 475 20413098
2010 The role of the kinases RIP1 and RIP3 in TNF-induced necrosis. Science signaling 381 20354226
2019 Intratumoral activation of the necroptotic pathway components RIPK1 and RIPK3 potentiates antitumor immunity. Science immunology 327 31227597
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2014 RIPK1 both positively and negatively regulates RIPK3 oligomerization and necroptosis. Cell death and differentiation 244 24902904
2016 Cytotoxicity of crystals involves RIPK3-MLKL-mediated necroptosis. Nature communications 228 26817517
2020 RIPK3 Orchestrates Fatty Acid Metabolism in Tumor-Associated Macrophages and Hepatocarcinogenesis. Cancer immunology research 224 32122992
2014 The necroptosis adaptor RIPK3 promotes injury-induced cytokine expression and tissue repair. Immunity 221 25367573
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2016 RIPK3 Restricts Myeloid Leukemogenesis by Promoting Cell Death and Differentiation of Leukemia Initiating Cells. Cancer cell 174 27411587
2017 Complex Pathologic Roles of RIPK1 and RIPK3: Moving Beyond Necroptosis. Trends in pharmacological sciences 155 28126382
2017 RIPK3 Restricts Viral Pathogenesis via Cell Death-Independent Neuroinflammation. Cell 152 28366204
2022 Roles of RIPK3 in necroptosis, cell signaling, and disease. Experimental & molecular medicine 147 36224345
2016 Contribution of RIP3 and MLKL to immunogenic cell death signaling in cancer chemotherapy. Oncoimmunology 144 27471616
2020 Inhibitors Targeting RIPK1/RIPK3: Old and New Drugs. Trends in pharmacological sciences 140 32035657
2018 Nuclear RIPK3 and MLKL contribute to cytosolic necrosome formation and necroptosis. Communications biology 135 30271893
2015 Ppm1b negatively regulates necroptosis through dephosphorylating Rip3. Nature cell biology 131 25751141
2014 RIPK1- and RIPK3-induced cell death mode is determined by target availability. Cell death and differentiation 129 24902899
2019 Pathogenesis of lupus nephritis: RIP3 dependent necroptosis and NLRP3 inflammasome activation. Journal of autoimmunity 128 31133359
2013 Positive and negative phosphorylation regulates RIP1- and RIP3-induced programmed necrosis. The Biochemical journal 118 24059293
2014 RIP3 is downregulated in human myeloid leukemia cells and modulates apoptosis and caspase-mediated p65/RelA cleavage. Cell death & disease 112 25144719
2018 Resibufogenin suppresses colorectal cancer growth and metastasis through RIP3-mediated necroptosis. Journal of translational medicine 108 30029665
2018 A RIPK3-PGE2 Circuit Mediates Myeloid-Derived Suppressor Cell-Potentiated Colorectal Carcinogenesis. Cancer research 103 30012671
2015 RIP3-mediated necrotic cell death accelerates systematic inflammation and mortality. Proceedings of the National Academy of Sciences of the United States of America 92 26283397
2018 PELI1 Selectively Targets Kinase-Active RIP3 for Ubiquitylation-Dependent Proteasomal Degradation. Molecular cell 91 29883609
2022 Mosaic composition of RIP1-RIP3 signalling hub and its role in regulating cell death. Nature cell biology 87 35256774
2016 The necroptosis-inducing kinase RIPK3 dampens adipose tissue inflammation and glucose intolerance. Nature communications 84 27323669
2021 Conformational interconversion of MLKL and disengagement from RIPK3 precede cell death by necroptosis. Nature communications 83 33850121
2021 A specific RIP3+ subpopulation of microglia promotes retinopathy through a hypoxia-triggered necroptotic mechanism. Proceedings of the National Academy of Sciences of the United States of America 79 33836603
2016 The Inflammatory Signal Adaptor RIPK3: Functions Beyond Necroptosis. International review of cell and molecular biology 79 28069136
2021 Human RIPK3 maintains MLKL in an inactive conformation prior to cell death by necroptosis. Nature communications 77 34811356
2017 RIP3 AND pMLKL promote necroptosis-induced inflammation and alter membrane permeability in intestinal epithelial cells. Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver 75 28844856
2018 BRAF and AXL oncogenes drive RIPK3 expression loss in cancer. PLoS biology 73 30157175
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2022 Human ZBP1 induces cell death-independent inflammatory signaling via RIPK3 and RIPK1. EMBO reports 68 36268590
2017 RIP1-RIP3-DRP1 pathway regulates NLRP3 inflammasome activation following subarachnoid hemorrhage. Experimental neurology 68 28579326
2013 More to life than death: molecular determinants of necroptotic and non-necroptotic RIP3 kinase signaling. Current opinion in immunology 67 24556404
2020 Bisphenol A induces coronary endothelial cell necroptosis by activating RIP3/CamKII dependent pathway. Scientific reports 63 32144343
2019 Necroptosis mediators RIPK3 and MLKL suppress intracellular Listeria replication independently of host cell killing. The Journal of cell biology 63 30975711
2020 Catalytically inactive RIP1 and RIP3 deficiency protect against acute ischemic stroke by inhibiting necroptosis and neuroinflammation. Cell death & disease 60 32703968
2016 Regulation of RIPK3- and RHIM-dependent Necroptosis by the Proteasome. The Journal of biological chemistry 60 26786097
2021 A toolbox for imaging RIPK1, RIPK3, and MLKL in mouse and human cells. Cell death and differentiation 59 33589776
2020 RIP-roaring inflammation: RIPK1 and RIPK3 driven NLRP3 inflammasome activation and autoinflammatory disease. Seminars in cell & developmental biology 57 32771377
2014 Rip3 knockdown rescues photoreceptor cell death in blind pde6c zebrafish. Cell death and differentiation 56 24413151
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2016 RIP3 induces ischemic neuronal DNA degradation and programmed necrosis in rat via AIF. Scientific reports 54 27377128
2017 Kinase Activities of RIPK1 and RIPK3 Can Direct IFN-β Synthesis Induced by Lipopolysaccharide. Journal of immunology (Baltimore, Md. : 1950) 53 28461567
2022 Caspase-8 auto-cleavage regulates programmed cell death and collaborates with RIPK3/MLKL to prevent lymphopenia. Cell death and differentiation 51 35064213
2020 RIPK3-mediated inflammation is a conserved β cell response to ER stress. Science advances 51 33355143
2019 Ptpn6 inhibits caspase-8- and Ripk3/Mlkl-dependent inflammation. Nature immunology 50 31819256
2020 RIPK3 upregulation confers robust proliferation and collateral cystine-dependence on breast cancer recurrence. Cell death and differentiation 48 31988496
2020 Somatic Epigenetic Silencing of RIPK3 Inactivates Necroptosis and Contributes to Chemoresistance in Malignant Mesothelioma. Clinical cancer research : an official journal of the American Association for Cancer Research 45 33203643
2024 Sensing of mitochondrial DNA by ZBP1 promotes RIPK3-mediated necroptosis and ferroptosis in response to diquat poisoning. Cell death and differentiation 44 38493248
2021 E3 ligase TRIM25 ubiquitinates RIP3 to inhibit TNF induced cell necrosis. Cell death and differentiation 44 33953350
2019 RIP3 deficiency alleviates liver fibrosis by inhibiting ROCK1-TLR4-NF-κB pathway in macrophages. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 44 31295018
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2022 RIPK3 dampens mitochondrial bioenergetics and lipid droplet dynamics in metabolic liver disease. Hepatology (Baltimore, Md.) 43 36029129
2021 Surviving death: emerging concepts of RIPK3 and MLKL ubiquitination in the regulation of necroptosis. The FEBS journal 43 34710282
2019 Dectin-1-induced RIPK1 and RIPK3 activation protects host against Candida albicans infection. Cell death and differentiation 43 30944411
2017 RIPK3 Mediates Necroptosis during Embryonic Development and Postnatal Inflammation in Fadd-Deficient Mice. Cell reports 42 28445730
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2023 OASL phase condensation induces amyloid-like fibrillation of RIPK3 to promote virus-induced necroptosis. Nature cell biology 37 36604592
2022 Inactivation of RIP3 kinase sensitizes to 15LOX/PEBP1-mediated ferroptotic death. Redox biology 37 35101798
2022 RIP3 Translocation into Mitochondria Promotes Mitofilin Degradation to Increase Inflammation and Kidney Injury after Renal Ischemia-Reperfusion. Cells 37 35741025
2019 Ablation of RIP3 protects from dopaminergic neurodegeneration in experimental Parkinson's disease. Cell death & disease 36 31690718
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2021 RIPK3 Activates MLKL-mediated Necroptosis and Inflammasome Signaling during Streptococcus Infection. American journal of respiratory cell and molecular biology 35 33625952
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2015 The serine threonine kinase RIP3: lost and found. BMB reports 35 25858093
2023 RIPK1 blocks T cell senescence mediated by RIPK3 and caspase-8. Science advances 34 36696505
2022 Bone Marrow-Derived RIPK3 Mediates Kidney Inflammation in Acute Kidney Injury. Journal of the American Society of Nephrology : JASN 33 35046131
2022 RIPK3 inhibitor-AZD5423 alleviates acute kidney injury by inhibiting necroptosis and inflammation. International immunopharmacology 32 36166972
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2022 Mechanisms of TNF-independent RIPK3-mediated cell death. The Biochemical journal 31 36240069
2019 Casein kinase-1γ1 and 3 stimulate tumor necrosis factor-induced necroptosis through RIPK3. Cell death & disease 31 31801942
2023 RIPK3-MLKL necroptotic signalling amplifies STING pathway and exacerbates lethal sepsis. Clinical and translational medicine 30 37475188
2023 PRMT1 reverts the immune escape of necroptotic colon cancer through RIP3 methylation. Cell death & disease 28 37005412
2023 Inhibition of gingival fibroblast necroptosis mediated by RIPK3/MLKL attenuates periodontitis. Journal of clinical periodontology 28 37366309
2019 RIP1, RIP3, and MLKL Contribute to Cell Death Caused by Clostridium perfringens Enterotoxin. mBio 26 31848291
2019 Inflammation, necrosis, and the kinase RIP3 are key mediators of AAG-dependent alkylation-induced retinal degeneration. Science signaling 25 30755477
2020 Cell-specific and athero-protective roles for RIPK3 in a murine model of atherosclerosis. Disease models & mechanisms 24 31953345
2023 Absence of Either Ripk3 or Mlkl Reduces Incidence of Hepatocellular Carcinoma Independent of Liver Fibrosis. Molecular cancer research : MCR 23 37204757
2021 A phosphorylation of RIPK3 kinase initiates an intracellular apoptotic pathway that promotes prostaglandin2α-induced corpus luteum regression. eLife 23 34029184
2016 RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death. Biochimica et biophysica acta. Molecular cell research 23 27984090
2020 RIPK3-MLKL-Mediated Neutrophil Death Requires Concurrent Activation of Fibroblast Activation Protein-α. Journal of immunology (Baltimore, Md. : 1950) 22 32796025
2021 MLKL and CaMKII Are Involved in RIPK3-Mediated Smooth Muscle Cell Necroptosis. Cells 21 34572045
2020 Enhanced RIPK3 kinase activity-dependent lytic cell death in M1 but not M2 macrophages. Molecular immunology 21 33221042
2018 RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells. Neuropsychiatric disease and treatment 21 30532542
2024 Expression of HIF1α in intestinal epithelium restricts arthritis inflammation by inhibiting RIPK3-induced cell death machinery. Annals of the rheumatic diseases 20 38503474
2024 RIPK3 causes mitochondrial dysfunction and albuminuria in diabetic podocytopathy through PGAM5-Drp1 signaling. Metabolism: clinical and experimental 20 39089491
2022 Curcumin Blocks High Glucose-Induced Podocyte Injury via RIPK3-Dependent Pathway. Frontiers in cell and developmental biology 20 35706905
2022 RIPK3-MLKL signaling activates mitochondrial CaMKII and drives intrarenal extracellular matrix production during CKD. Matrix biology : journal of the International Society for Matrix Biology 20 35964866
2022 RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity. Molecular metabolism 20 36030035