DTX3L induced NLRP3 ubiquitination inhibit R28 cell pyroptosis in OGD/R injury

Ischemia/reperfusion (I/R) injury is one of the most common etiologies in many diseases. Retinal I/R leads to cytokine storm, resulting in tissue damage and cell death. Pyroptosis, a novel type of regulated cell death, occurs after cellular I/R injury. In this study, we established an oxygen glucose...

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Bibliographic Details
Published in:Biochimica et biophysica acta. Molecular cell research 2023-03, Vol.1870 (3), p.119433-119433, Article 119433
Main Authors: Zhou, Ziyu, Shang, Lei, Zhang, Qi, Hu, Ximin, Huang, Ju-fang, Xiong, Kun
Format: Article
Language:English
Subjects:
Cell Death
Cell Line
DTX3L
Humans
NLR Family, Pyrin Domain-Containing 3 Protein - genetics
NLR Family, Pyrin Domain-Containing 3 Protein - metabolism
NLRP3
OGD/R
Pyroptosis
Reperfusion Injury - metabolism
Ubiquitin-Protein Ligases
Ubiquitination
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Summary:Ischemia/reperfusion (I/R) injury is one of the most common etiologies in many diseases. Retinal I/R leads to cytokine storm, resulting in tissue damage and cell death. Pyroptosis, a novel type of regulated cell death, occurs after cellular I/R injury. In this study, we established an oxygen glucose deprivation (OGD/R) cellular model (R28) to simulate retinal I/R injury. We conducted an LDH assay, and EthD-III and PI staining procedures to confirm pyroptosis. Mass spectrometry and bioinformatics analysis were used to identify the possible proteins interacting with NLRP3. Co-IP and various molecular biology techniques were used to investigate the possible modes regulating NLRP3 by DTX3L. EthD-III, PI staining and LDH assays demonstrated pyroptosis induced by OGD/R injury, mediated via NLRP3 pathway. Mass spectrometry and bioinformatics analysis screened out three candidate proteins interacting with NLRP3, and further Co-IP experiment indicated that DTX-3L may interact with NLRP3 to regulate its protein levels after injury. Co-IP experiments and various molecular biology methods demonstrated that DTX3L ubiquitinates NLRP3 resulting in pyroptosis after R28 OGD/R injury. Further, NLRP3 LRR and DTX3L RING domains interact with each other. Our study demonstrated that DTX3L may ubiquitinate NLRP3 to regulate OGD/R-induced pyroptosis globally in R28 cells. •DTX3L may ubiquitinate NLRP3 to regulate OGD/R-induced pyroptosis in R28 cells.•The RING domain of DTX3L is the key that regulates NLRP3 ubiquitination.•LRR domain of NLRP3 is the key for DTX3L binding via K48 site.
ISSN:0167-4889
1879-2596
DOI:10.1016/j.bbamcr.2023.119433