| 1996 |
TRAF5 was identified as a novel TRAF family member containing RING finger, zinc finger, coiled-coil, and TRAF homology domains. In vitro translation and co-immunoprecipitation in COS7 cells showed TRAF5 binds the cytoplasmic region of the lymphotoxin-beta receptor (LT-βR) but not CD40, both TNF receptors, Fas, or NGF receptor. Overexpression of full-length TRAF5 (but not a truncated form lacking the zinc-binding region) activated NF-κB in HEK293 cells, and dominant-negative TRAF5 partially inhibited LT-βR-induced NF-κB activation. |
In vitro binding assay (translated protein), co-immunoprecipitation in COS7 cells, NF-κB reporter assay in HEK293 cells, domain truncation analysis |
The Journal of biological chemistry |
High |
8663299
|
| 1996 |
TRAF5 was cloned via yeast two-hybrid using the CD40 cytoplasmic tail as bait. In vitro binding assays confirmed TRAF5 associates with CD40 (residues 230–269 required) but not TNFR2. Overexpression of TRAF5 activates NF-κB, and amino-terminally truncated TRAF5 suppresses CD40-mediated CD23 induction, similar to TRAF3. |
Yeast two-hybrid, in vitro binding assay, NF-κB reporter assay, CD40 deletion mutant analysis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
8790348
|
| 1997 |
Human TRAF5 cDNA was cloned and mapped to chromosome 1q32. The encoded 557-aa protein retains typical TRAF family structural features. Overexpression of human TRAF5 activated NF-κB in 293T cells, and the protein was found to bind LT-βR cytoplasmic region more efficiently than CD40 or CD30. |
cDNA cloning, chromosomal mapping (FISH), NF-κB reporter assay, binding assay |
Genomics |
Medium |
9177772
|
| 1997 |
The novel TNFR family member ATAR (both human and mouse) physically interacts with TRAF5 and TRAF2 via its C-terminal 20 amino acids. Co-expression of ATAR with TRAF5 (but not TRAF2) results in synergistic NF-κB activation, indicating differential roles of TRAF2 and TRAF5 downstream of ATAR. |
In vitro binding assay, co-immunoprecipitation, NF-κB reporter assay |
The Journal of biological chemistry |
Medium |
9153189
|
| 1998 |
CD27 activates NF-κB and SAPK/JNK through direct interaction with TRAF2 and TRAF5 via its C-terminal PIQEDYR motif. Dominant-negative TRAF2 or TRAF5 blocked both NF-κB and SAPK/JNK activation induced by CD27. NF-κB-inducing kinase (NIK) acts as a common downstream kinase of TRAF2 and TRAF5 in this pathway. |
Cytoplasmic domain deletion analysis, dominant-negative transfection, NF-κB EMSA, kinase assay |
The Journal of biological chemistry |
High |
9582383
|
| 1998 |
OX40 associates with TRAF1, TRAF2, TRAF3, and TRAF5 (but not TRAF4) in vitro, and with TRAF2, TRAF3, and TRAF5 in vivo. A cytoplasmic sequence (aa 256–263, GGSFRTPI) is required for TRAF association and NF-κB activation. Dominant-negative TRAF2 and TRAF5 suppress OX40-induced NF-κB activation in a dose-dependent manner. |
GST pull-down, co-immunoprecipitation in HEK293T, NF-κB EMSA, deletion mutant analysis, dominant-negative transfection |
The Journal of biological chemistry |
High |
9488716
|
| 1998 |
Human TRAF5 protein binds the LT-βR cytoplasmic region more efficiently than CD40 or CD30, and overexpression activates NF-κB in 293T cells. The gene was mapped to human chromosome 1q32.3–q41.1. |
cDNA cloning, binding assay, NF-κB reporter assay, chromosomal mapping (PCR-RFLP) |
Gene |
Medium |
9511754
|
| 1999 |
TRAF5-deficient mice generated by gene targeting showed that TRAF5 loss does not completely abrogate TNF-, CD27-, or CD40-induced NF-κB or JNK activation, but TRAF5-/- B cells exhibit defects in CD40-driven proliferation, upregulation of CD23, CD54, CD80, CD86, and Fas, and reduced IgG production with IL-4. TRAF5-/- T cells show impaired CD27-mediated costimulatory signaling. |
Gene targeting (KO mice), NF-κB EMSA, JNK kinase assay, flow cytometry, in vitro Ig production assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
10449775
|
| 2001 |
TRAF2 and TRAF5 double knockout (DKO) MEFs show severely impaired TNF-induced (but not IL-1-induced) NF-κB nuclear translocation, establishing redundant roles for TRAF2 and TRAF5 specifically in TNF-induced NF-κB activation. DKO MEFs are more susceptible to TNF-induced cytotoxicity than TRAF2 single KO cells. |
Double knockout mouse embryonic fibroblasts, NF-κB nuclear translocation assay, cytotoxicity assay |
The Journal of biological chemistry |
High |
11479302
|
| 2001 |
Overexpression of TRAF5 (and TRAF6), or LMP1 (via its TRAF-binding site), suppresses Epstein-Barr virus oriP replication through a p38 MAPK-dependent pathway. Dominant-negative TRAF5 and TRAF6 relieve LMP1-induced oriP suppression; p38 MAPK inhibition abolishes the suppressive effect. |
Transient replication assay, dominant-negative transfection, p38 MAPK inhibitor, LMP1 deletion mutant analysis |
Journal of virology |
Medium |
11333886
|
| 2002 |
In Hodgkin-Reed-Sternberg (H-RS) cells with constitutively active CD30 signaling, TRAF2 and TRAF5 aggregate in cytoplasmic clusters and co-localize with IKKα, NIK, and IκBα. Dominant-negative TRAF2 and TRAF5 suppressed cytoplasmic aggregation and constitutive NF-κB activation, suggesting TRAF5 functions as a scaffolding protein in CD30-driven NF-κB signaling. |
Confocal immunofluorescence microscopy, dominant-negative transfection, NF-κB assay |
The American journal of pathology |
Medium |
12000717
|
| 2003 |
TNF-α-induced phosphorylation of NF-κB p65 on Ser-536 is mediated through a TRAF2/TRAF5–TAK1–IKK pathway. This phosphorylation is severely impaired in MEFs from traf2-/-traf5-/- double KO mice. Dominant-negative TAK1, IKKα, IKKβ, and siRNAs against TAK1, IKKα, IKKβ each blocked the phosphorylation. |
Anti-phospho-p65 (Ser-536) antibody, double KO MEFs, dominant-negative overexpression, siRNA knockdown |
The Journal of biological chemistry |
High |
12842894
|
| 2003 |
TRAF5-deficient osteoclast progenitor cells fail to differentiate effectively into mature multinucleated osteoclasts in response to RANKL or TNFα, even though JNK and NF-κB activation is preserved, demonstrating TRAF5 is required for osteoclastogenesis downstream of or parallel to these signaling events. In vivo, PTH-induced hypercalcemia is delayed in TRAF5-deficient mice. |
TRAF5-deficient mouse osteoclast progenitor cultures, RANKL/TNFα stimulation, JNK/NF-κB activation assay, PTH hypercalcemia model |
Journal of bone and mineral research |
High |
12619928
|
| 2009 |
TRAF5 associates strongly with Epstein-Barr virus latent membrane protein 1 (LMP1) — more strongly than with CD40 — and is required for LMP1-mediated c-Jun kinase signaling and B cell hyperactivation phenotypes. In mice expressing LMP1 in place of CD40, TRAF5 deficiency abrogated much of the abnormal splenic phenotype (splenomegaly, lymphadenopathy, elevated IL-6, autoantibodies). |
Co-immunoprecipitation, LMP1/CD40 transgenic mice crossed with TRAF5-KO mice, JNK assay, in vivo phenotyping |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19805155
|
| 2009 |
In TRAF2/TRAF5 double KO (T2/5 DKO) cells, basal IKK activity and NF-κB-dependent gene expression are elevated (not reduced) due to elevated NIK activity. TNFα-induced RIP1 ubiquitination is impaired in DKO cells, yet TNFα can still further activate IKK. TRAF2 (not TRAF5) is required for recruiting anti-apoptotic proteins to TNFR1 complex and protecting against TNFα-induced cell death. |
DKO MEF cells, IKK kinase assay, NF-κB target gene expression, NIK inhibition, TNFR1 complex immunoprecipitation, cytotoxicity assay |
Journal of molecular biology |
High |
19409903
|
| 2010 |
TRAF5 is a downstream effector of MAVS in antiviral innate immune signaling. The MAVS transmembrane domain mediates dimerization and subsequent association with TRAF5 and induction of TRAF5 ubiquitination in a CARD-dependent manner. TRAF5 mediates both IRF3 and NF-κB activation downstream of MAVS, and NEMO is recruited to dimerized MAVS in a TRAF3/TRAF5-dependent manner. |
Co-immunoprecipitation, ubiquitination assay, IRF3/NF-κB reporter assays, MAVS domain truncation, siRNA knockdown of TRAF5 |
PloS one |
Medium |
20161788
|
| 2010 |
TRAF5 deficiency in mice accelerates atherosclerosis: TRAF5-/-/LDLR-/- mice develop larger lesions with more lipids and macrophages. TRAF5 deficiency in endothelial cells or leukocytes enhances inflammatory cell adhesion and increases JNK activation, and TRAF5-deficient macrophages show enhanced foam cell formation. These effects appear independent of TRAF2. |
TRAF5/LDLR double KO mice on high-cholesterol diet, intravital microscopy, dynamic adhesion assays, foam cell lipid uptake assay, JNK activation measurement |
Circulation research |
High |
20651286
|
| 2011 |
IL-17 stabilizes CXCL1 mRNA via a pathway involving Act1→TRAF2/TRAF5→SF2(ASF). TRAF2 and TRAF5 are necessary for IL-17-induced mRNA stabilization. IL-17 promotes formation of a TRAF5–TRAF2–Act1–SF2(ASF) complex; SF2(ASF) binding to CXCL1 mRNA is reduced after IL-17 stimulation, correlating with mRNA stabilization. |
siRNA knockdown, mRNA half-life assay, co-immunoprecipitation, RNA-protein binding assay |
Nature immunology |
High |
21822258
|
| 2012 |
Numbl directly interacts with TRAF5 and promotes K48-linked polyubiquitination of TRAF5, committing it to proteasomal degradation, thereby dampening TRAF5-dependent NF-κB activation and inhibiting glioma cell migration and invasion. |
Co-immunoprecipitation, ubiquitination assay (K48-specific), proteasome inhibitor treatment, overexpression/knockdown of Numbl, NF-κB reporter, migration/invasion assays |
Molecular biology of the cell |
Medium |
22593207
|
| 2012 |
Crystal structures of the TRAF domain of TRAF5 and TRAF3 (bound to a Cardif/MAVS TRAF-interacting motif peptide) were solved. Structural comparison identified two key residues in TRAF3 (Tyr440 and Phe473) near the Cardif binding pocket absent in TRAF5. Mutating the corresponding TRAF5 residues to match TRAF3 conferred TRAF3-like antiviral (IFN-inducing) activity on TRAF5. |
X-ray crystallography, in vitro binding assay, cellular IFN reporter assay, site-directed mutagenesis |
Science signaling |
High |
23150880
|
| 2013 |
TRAF5 is a negative regulator of TLR signaling in B lymphocytes. TRAF5-/- B cells overproduce IL-6, IL-12p40, IL-10, TNF-α, and IgM upon TLR stimulation, with markedly enhanced phosphorylation of ERK1/2 and JNK but no effect on NF-κB or cell survival. Following TLR stimulation, TRAF5 associates with MyD88 and TAB2, and negatively regulates TAB2–TRAF6 association. |
TRAF5 KO B cells, cytokine ELISA, flow cytometry, phospho-MAPK western blot, co-immunoprecipitation (TRAF5 with MyD88/TAB2/TRAF6) |
Journal of immunology |
High |
24259503
|
| 2014 |
TRAF5 constitutively associates with a cytoplasmic region of gp130 that overlaps the STAT3 binding site, suppressing STAT3 recruitment and activation in response to IL-6, thereby limiting Th17 differentiation. TRAF5-deficient naïve CD4+ T cells show enhanced Th17 differentiation and TH17-driven EAE is greatly exacerbated in Traf5-/- mice. |
Co-immunoprecipitation (TRAF5–gp130), STAT3 phosphorylation assay, Traf5-/- mouse model, Th17 differentiation assay, EAE model |
Nature immunology |
High |
24681564
|
| 2015 |
TRAF5 directly interacts with RORγt (the Th17 master transcription factor) and promotes K63-linked polyubiquitination of RORγt via its RING finger domain, stabilizing RORγt protein. TRAF5 depletion in Th17 cells destabilizes RORγt and downregulates IL-17A and other Th17-related genes. |
Co-immunoprecipitation, ubiquitination assay (K63-specific), RING domain mutant, TRAF5 knockdown in Th17 cells, RORγt protein stability assay, qRT-PCR |
The Journal of biological chemistry |
High |
26453305
|
| 2016 |
TRAF5 negatively regulates NAFLD/NASH by blocking JNK1 (but not JNK2) activity. TRAF5 deficiency worsens HFD-induced metabolic disorders, and Jnk1 ablation markedly ameliorates the detrimental effects of Traf5 deficiency on obesity, inflammation, insulin resistance, hepatic steatosis, and fibrosis. |
TRAF5 KO and overexpression in mice (HFD and ob/ob models), JNK1/JNK2-selective genetic ablation, liver histology, metabolic parameters |
Journal of hepatology |
High |
27032381
|
| 2016 |
Deep mutational scanning of TRAF2, TRAF3, and TRAF5 MATH domain peptide-binding preferences revealed that TRAF5 shows distinct peptide binding specificity compared to TRAF2 and TRAF3. Different preferences were identified in both CD40 and TANK background peptide libraries, demonstrating previously unappreciated binding selectivity among TRAF paralogs. |
Deep mutational scanning, bacterial surface display of peptide libraries, next-generation sequencing enrichment analysis, individual peptide affinity measurement |
Protein science |
Medium |
26779844
|
| 2018 |
TRAF2 and TRAF5 constitutively bind to gp130 and inhibit IL-6-driven JAK1 transphosphorylation by limiting proximal JAK1–JAK1 interaction in the IL-6 receptor complex. This was demonstrated using a luciferase fragment complementation system for JAK1–JAK1 proximity. Traf5-/- CD4+ T cells display significantly higher IL-6-induced pJAK1 levels than wild-type cells. |
Luciferase fragment complementation for JAK1–JAK1 interaction, HEK293T co-transfection, phospho-JAK1 western blot in Traf5-/- T cells |
International immunology |
Medium |
29668931
|
| 2019 |
TRAF5 promotes plasmacytoid dendritic cell (pDC) development from bone marrow progenitors in a cell-intrinsic manner. TRAF5 regulates the balance of transcription factors TCF4 and ID2 to promote pDC versus conventional DC commitment. |
TRAF5-/- mice, bone marrow chimera experiments, flow cytometry of DC subsets, bone marrow progenitor culture, TCF4/ID2 expression analysis |
Biochemical and biophysical research communications |
Medium |
31668809
|
| 2020 |
14-3-3ζ physically interacts with TRAF5 (and TRAF6), and this interaction is increased in the presence of IL-17A. TRAF5 acts as an endogenous suppressor of IL-17A-induced IL-6 production, and 14-3-3ζ counters TRAF5's suppressive effect. 14-3-3ζ interaction with TRAF proteins is required for IL-17A-induced IL-6 levels, while TRAF5 and TRAF6 define distinct branches (IL-6 vs. CXCL-1) of IL-17A signaling. |
Co-immunoprecipitation, genetically manipulated human and mouse cells, ex vivo and in vivo rat models, IL-6/CXCL-1 production assay |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
32968020
|
| 2020 |
TRAF5 protects against myocardial ischemia/reperfusion injury by promoting AKT activation. TRAF5-knockout mice exhibit heavier heart damage, inflammatory response, and cell death after I/R injury. TRAF5 overexpression in H/R-stimulated cardiomyocytes inhibits inflammation and apoptosis. |
TRAF5 KO mice (myocardial I/R model), cardiomyocyte overexpression, AKT phosphorylation assay, apoptosis/inflammation readouts |
European journal of pharmacology |
Medium |
32234528
|
| 2020 |
TRAF5 deficiency in nonhematopoietic intestinal epithelial cells reduces TRAF2 protein stability in a proteasome-dependent manner during inflammation, identifying TRAF5 as required for TRAF2 protein maintenance in inflamed colon tissue. |
TRAF5 KO mice (DSS colitis model), bone marrow chimeras, TRAF2 protein assay, proteasome inhibitor treatment, proinflammatory cytokine stimulation of TRAF5-/- nonhematopoietic cells |
ImmunoHorizons |
Medium |
32156688
|
| 2021 |
TRAF5 deficiency in mice aggravates diet-induced obesity and metabolic derangements. TRAF5-deficient adipocytes (but not leukocytes) show increased expression of TNFα, MIP-1α, MCP-1, and RANTES, identifying TRAF5 as an anti-inflammatory regulator in adipocytes. |
Traf5-/- mice on high-fat diet, flow cytometry of adipose stromal vascular fraction, cell-type-specific gene expression, adipocyte isolation |
Arteriosclerosis, thrombosis, and vascular biology |
Medium |
34348490
|
| 2021 |
TRAF5 silencing in HCC cells enhances necroptosis by suppressing LTBR (lymphotoxin-beta receptor)-mediated NF-κB signaling. Co-immunoprecipitation confirmed interaction between TRAF5 and LTBR. LTBR overexpression abolished the promotive effect of TRAF5 knockdown on necroptosis and reversed NF-κB suppression. |
Co-immunoprecipitation (TRAF5–LTBR), siRNA knockdown, LTBR overexpression rescue, NF-κB western blot, necroptosis markers (p-RIP1, p-MLKL), xenograft model |
PeerJ |
Medium |
37366426
|
| 2023 |
TRAF5 regulates intestinal Th1/Th17 cell differentiation through Runx1. In a T-cell transfer colitis model, TRAF5-/- CD4+ T cells cause more severe colitis with increased IFN-γ, TNF-α, IL-17a. AAV-mediated Runx1 knockout inhibited TRAF5-/- CD4+ T cell differentiation into Th1 and Th17 cells in vivo, placing Runx1 downstream of TRAF5 in this pathway. |
T-cell transfer colitis model (Rag2-/- recipients), AAV-mediated Runx1 KO in vivo, flow cytometry, cytokine measurement (ELISA, qRT-PCR, IHC) |
Immunology |
Medium |
37575027
|
| 2024 |
IL-17A promotes formation of a TRAF2/TRAF5/HuR complex that enhances PFKFB3 mRNA stability and expression, thereby activating glycolysis in hepatic stellate cells. Co-IP, RNA immunoprecipitation, and RNA pull-down confirmed interactions among TRAF2, TRAF5, and HuR in this complex. Silencing TRAF2 or TRAF5 abolished IL-17A-induced PFKFB3 upregulation and glycolysis. |
Co-immunoprecipitation, RNA immunoprecipitation (RIP), RNA pull-down, siRNA knockdown, glycolysis assay (ECAR), western blot |
Central-European journal of immunology |
Medium |
39944257
|
| 2025 |
The E3 ubiquitin ligase Huwe1 is required for TRAF5 activity in type I IFN induction downstream of RIG-I-like receptors. Proteomics identified TRAF5 (and MAVS and other TRAFs) as putative Huwe1 substrates; TRAF5 physically interacts with Huwe1; and Huwe1 is essential for TRAF5-dependent IFN-β induction. |
CRISPR KO of Huwe1, IFN-β reporter assay, proteomics for substrate identification, co-immunoprecipitation (Huwe1–TRAF5), primary macrophages |
bioRxivpreprint |
Low |
bio_10.1101_2025.03.27.645708
|