Genomic locus proteomic screening identifies the NF-κB signaling pathway components NFκB1 and IKBKG as transcriptional regulators of Ripk3 in endothelial cells.
Genomic locus proteomic screening identifies the NF-κB signaling pathway components NFκB1 and IKBKG as transcriptional regulators of Ripk3 in endothelial cells.
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DOI：
10.1371/journal.pone.0253519
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Griffin CT
中科院分区：
文献类型：
--
作者：
Gao S;Menendez M;Kurylowicz K;Griffin CT
受体相互作用蛋白激酶3 （RIPK3）是一种多功能蛋白，以促进细胞坏死和炎症而闻名。我们实验室最近的证据表明，内皮细胞必须严格调节RIPK3的表达，以促进血管生成，在胚胎发生过程中保持血管完整性，并提供对出生后动脉粥样硬化的保护。RIPK3的活性和稳定性受翻译后修饰和caspase依赖性切割的调控。然而，对Ripk3的转录调控知之甚少。在这里，我们利用一种无偏倚的基于crispr的技术，称为基因组位点蛋白质组学（GLoPro）来筛选小鼠内皮细胞系中与Ripk3位点相关的转录因子和协调节蛋白。我们发现41种核蛋白在Ripk3位点特异性富集，包括活化B细胞的核因子kappa轻链增强子（NF-κB）信号通路成分NF-κ b1和IKBKG。在体外培养的人原代内皮细胞中，我们进一步验证了NFκB1和IKBKG直接结合Ripk3启动子，阻止tnf α诱导的Ripk3转录。此外，NFκB1可阻止ripk3介导的原代内皮细胞死亡。这些数据为内皮细胞中NF-κB信号传导和Ripk3转录调控提供了新的见解。
The receptor-interacting protein kinase 3 (RIPK3) is a multi-functional protein best known for facilitating cellular necroptosis and inflammation. Recent evidence from our lab indicates that RIPK3 expression must be tightly regulated in endothelial cells to promote angiogenesis, to maintain vascular integrity during embryogenesis, and to provide protection from postnatal atherosclerosis. RIPK3 activity and stability are regulated by post-translational modifications and caspase-dependent cleavage. However, less is known about the transcriptional regulation of Ripk3. Here we utilized an unbiased CRISPR-based technology called genomic locus proteomics (GLoPro) to screen transcription factors and coregulatory proteins associated with the Ripk3 locus in a murine endothelial cell line. We found that 41 nuclear proteins are specifically enriched at the Ripk3 locus, including the Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway components NFκB1 and IKBKG. We further verified that NFκB1 and IKBKG directly bind the Ripk3 promoter and prevent TNFα-induced Ripk3 transcription in cultured human primary endothelial cells. Moreover, NFκB1 prevents RIPK3-mediated death of primary endothelial cells. These data provide new insights into NF-κB signaling and Ripk3 transcriptional regulation in endothelial cells.
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影响因子：
39.3
作者：
Liu T;Zhang L;Joo D;Sun SC
通讯作者：
Sun SC
影响因子：
15.9
作者：
Kisseleva, Tatiana;Song, Li;Schindler, Christian
通讯作者：
Schindler, Christian
影响因子：
16
作者：
Choi, Seung-Won;Park, Han-Hee;Kim, You-Sun
通讯作者：
Kim, You-Sun
影响因子：
44.1
作者：
Koo, Gi-Bang;Morgan, Michael J.;Kim, You-Sun
通讯作者：
Kim, You-Sun
RIP3 的裂解通过去除激酶结构域使其不依赖 caspase 的细胞凋亡途径失活
影响因子：
4.8
作者：
Feng, Shanshan;Yang, Yonghui;Wu, Mian
通讯作者：
Wu, Mian