| 2016 |
SPATA2 was identified as a novel component of the TNF receptor signaling complex (TNF-RSC). The N-terminal PUB domain of SPATA2 interacts with the USP domain of CYLD, while the C-terminus of SPATA2 interacts with HOIP (the catalytically active component of LUBAC). SPATA2 is required for recruitment of CYLD to the TNF-RSC. Loss of SPATA2 augments NF-κB transcriptional activation and inhibits TNF-α-induced necroptosis. |
Quantitative mass spectrometry of TNF-RSC composition, Co-IP/pulldown domain mapping, siRNA knockdown with NF-κB and necroptosis readouts |
The EMBO journal |
High |
27307491
|
| 2016 |
SPATA2 is a constitutive direct binding partner of HOIP that bridges the interaction between CYLD and HOIP, enabling CYLD recruitment to TNFR1- and NOD2-signaling complexes. SPATA2 recruitment to these complexes is dependent on HOIP. Loss of SPATA2 abolishes CYLD recruitment and diminishes TNF-induced necroptosis, resembling loss of CYLD. |
Co-IP, pulldown, CRISPR/siRNA loss-of-function with necroptosis and NF-κB readouts in multiple signaling complexes |
Cell reports |
High |
27545878
|
| 2016 |
SPATA2 interacts with CYLD via its PUB domain, while a PUB interaction motif (PIM) of SPATA2 interacts with the PUB domain of HOIP. Beyond scaffolding, SPATA2 acts as an allosteric activator of the K63- and M1-deubiquitinase activity of CYLD. SPATA2 is required for TNF-induced complex II formation, caspase activation, and apoptosis, and substantially attenuates TNF-induced NF-κB and MAPK signaling. |
Mass spectrometry screen, Co-IP domain mapping, in vitro deubiquitinase activity assay, loss-of-function with NF-κB/MAPK/apoptosis readouts |
EMBO reports |
High |
27458237
|
| 2017 |
SPATA2-deficient cells show resistance to RIPK1-dependent apoptosis and necroptosis, and partial protection against RIPK1-independent apoptosis. SPATA2 deficiency promotes M1 ubiquitination of RIPK1, thereby inhibiting RIPK1 kinase activity. Biochemical in vitro evidence shows that the CYLD USP domain complexed with the SPATA2 PUB domain preferentially deubiquitinates M1 (linear) ubiquitin chains. SPATA2 deficiency also promotes MKK4 and JNK activation and cytokine production independently of RIPK1 kinase activity. |
SPATA2-deficient cell lines (genetic KO), in vitro deubiquitinase assay with M1 and K63 ubiquitin chains, RIPK1 ubiquitination biochemistry, cell death assays |
Genes & development |
High |
28701375
|
| 2019 |
SPATA2 recruits CYLD to the centrosome to deubiquitinate PLK4 (polo-like kinase 4). Deubiquitination of PLK4 facilitates its phosphorylation of NEK7 at Ser204, which attenuates the NEK7–NLRP3 interaction required for NLRP3 inflammasome activation. SPATA2 deficiency enhances NLRP3 inflammasome activity in macrophages and in a peritonitis animal model. |
SPATA2 KO macrophages, Co-IP of PLK4/CYLD/NEK7/NLRP3, in vitro ubiquitination/deubiquitination assay, phosphorylation site mutagenesis (NEK7 S204A), shRNA knockdown, animal peritonitis model |
The EMBO journal |
High |
31762063
|
| 2023 |
SPATA2 has CYLD-independent functions: SPATA2 competes with OTULIN for binding to HOIP via its PIM and zinc finger domain, thereby promoting LUBAC autoubiquitylation and attenuating LUBAC activity. Double knockout (Cyld-/-Spata2-/-) mice show highly penetrant perinatal lethality absent in single knockouts. Cyld-/-Spata2-/- cells show increased M1-linked TNFR1-SC ubiquitylation and elevated pro-inflammatory gene expression compared to either single KO, and this increased signaling depends on OTULIN. |
Double KO mice (CRISPR), OTULIN competition binding assay, M1-ubiquitin chain analysis, pro-inflammatory gene expression in fibroblasts/macrophages/intestinal epithelial cells, genetic epistasis with OTULIN |
Cell reports |
High |
36640323
|
| 2022 |
SPATA2 and CYLD inhibit STAT1 accumulation and activation (and subsequent CXCL10 expression) in colorectal cancer cells. At steady state, STAT1 is highly ubiquitinated in a SPATA2/CYLD-dependent manner, suggesting SPATA2/CYLD deubiquitinate STAT1 to regulate IFN-γ signaling. Tumor-specific deletion of SPATA2 or CYLD enhances anti-PD-1 response in vivo. |
Specific deletion of SPATA2/CYLD in human and mouse CRC cell lines, STAT1 ubiquitination analysis, CXCL10 expression measurement, in vivo anti-PD-1 response assay |
Frontiers in oncology |
Medium |
36531014
|
| 2022 |
The SPATA2/CYLD pathway contributes to doxorubicin-induced cardiomyocyte ferroptosis by enhancing ferritinophagy. SPATA2 knockdown reduced NCOA4 deubiquitination, attenuated ferritinophagy, and decreased ferroptosis. The pathway promotes NCOA4 deubiquitination (via CYLD), which stabilizes NCOA4 and drives ferritin degradation and intracellular iron overload. |
SPATA2 knockdown in cardiomyocytes, NCOA4 ubiquitination assay, Co-IP of SPATA2/CYLD interaction, ferroptosis markers (iron, ACSL4, LC3), in vivo mouse doxorubicin model |
Chemico-biological interactions |
Medium |
36195186
|
| 2025 |
SPATA2/CYLD pathway-dependent deubiquitylation of p53 promotes ferroptosis in rat heart after ischemia/reperfusion by stabilizing p53, which suppresses SLC7A11 transcription, reducing GPX4 levels. SPATA2 knockdown restored p53 ubiquitination, increased SLC7A11 and GPX4, and reduced ferroptosis. |
SPATA2 knockdown in H9c2 cells and rat I/R model, p53 ubiquitination assay, SLC7A11/GPX4 expression, lurasidone as SPATA2 inhibitor |
Heart, lung & circulation |
Medium |
40389334
|
| 2001 |
SPATA2 protein tagged with GFP is located in the nucleus when expressed in HLtat transfected cells. SPATA2 mRNA is expressed in testis and Sertoli cells, and FSH treatment of Sertoli cells induces changes in steady-state spata2 mRNA in a time-dependent manner. |
GFP-fusion protein expression and fluorescence localization, RT-PCR, FSH treatment of Sertoli cells |
Biochemical and biophysical research communications |
Low |
11322771
|
| 2017 |
SPATA2 global knockout in mice (via CRISPR/Cas9n) results in approximately 40% decrease in testis size/weight, 40% decrease in sperm count, 28% decrease in germ cell proliferation, and approximately 70% increase in inhibin alpha-subunit mRNA and protein in testes, indicating SPATA2 ensures normal secretory function of Sertoli cells and is required for male fertility. |
CRISPR/Cas9n global KO mice, histomorphology, sperm count, BrdU proliferation assay, RT-PCR and Western blot for inhibin alpha |
Biology of reproduction |
Medium |
29025062
|
| 2023 |
SPATA2 suppresses β-catenin signaling by attenuating DVL1 ubiquitination. Ectopic overexpression of SPATA2 inhibited EMT, impaired motility and invasiveness in vitro, reduced metastasis in vivo, and increased radiosensitivity in NSCLC cells. |
SPATA2 overexpression/KD in NSCLC cell lines, DVL1 ubiquitination assay, β-catenin signaling reporter, in vitro invasion/migration assays, in vivo metastasis model |
Thoracic cancer |
Low |
36814090
|