| 朱翔,曹宝山,张照辉,等.TAK1赖氨酸158位点发生Lys 63连接多聚泛素化在辐射诱导NF-κB激活过程中的作用[J].中华放射医学与防护杂志,2013,33(3):235-238.ZHU Xiang,CAO Bao-shan,ZHANG Zhao-hui,et al.Roel of Lys63-linked TAK1 poly-ubiquitination at lysine 158 in radiation-induced NF-κB activity[J].Chin J Radiol Med Prot,2013,33(3):235-238 |
| TAK1赖氨酸158位点发生Lys 63连接多聚泛素化在辐射诱导NF-κB激活过程中的作用 |
| Roel of Lys63-linked TAK1 poly-ubiquitination at lysine 158 in radiation-induced NF-κB activity |
| 投稿时间：2013-02-19 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.03.004 |
| 中文关键词: 辐射损伤 转化生长因子β激酶1 多聚泛素化 NF-κB 信号通路 |
| 英文关键词:Radiation effect Transforming growth factor-β-activating kinase 1 Polyubiquitination Nuclear factor-kappa B Signaling pathway |
| 基金项目:卫生行业科研专项(201002009) |
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| 中文摘要: |
| 目的 研究转化生长因子激酶1(TAK1)Lys 63连接多聚泛素化在辐射导致的NF-κB激活过程中起到的作用及其作用位点。方法 通过基因瞬时转染技术将FLAG-TAK1和HA-Ub-K63质粒共同转染至HEK-293T细胞,应用免疫沉淀和免疫印迹技术研究辐射对TAK1泛素化的影响;用稳定转染TAK1表达的HeLa细胞,证实辐射对TAK1泛素化的影响。通过免疫印迹技术,比较分析辐射对稳定转染TAK1 K158R的MEF细胞与TAK1野生型MEF细胞比较分析辐射对它表达IKKs、p38及JNK的影响,以证明辐射诱导的TAK1-Lys 63多聚泛素化的位点是否在TAK1赖氨酸158位点。结果 瞬时转染的HEK-293T细胞接受辐射后1 h开始出现明显的TAK1泛素化表现,2 h更明显。在稳定表达FLAG-TAK1的 HeLa细胞系中得到证实。与TAK1野生型MEF细胞相比,TAK1 K158R突变的MEF细胞中辐射诱导的IKKs和 p38 磷酸化作用减低。结论 TAK1多聚泛素化在辐射导致的NF-κB激活过程中起到关键作用,辐射所致TAK1-Lys 63连接多聚泛素化发生在TAK1的赖氨酸158位点。 |
| 英文摘要: |
| Objective To study the role of the transforming growth factor-β-activating kinase 1 (TAK1) poly-ubiquitination in radiation-induced NF-κB activation.Methods FLAG-TAK1 and HA-Ub-K63 plasmids were transiently transfected into HEK-293T cells. The radiation effect on TAK1 ubiquitination was detected by immunoprecipitation and Western blot, which was further confirmed in FLAG-TAK1 HeLa stable cell line. Results At 1 h after irradiation, Lys 63-link TAK1 ubiquitination was induced in the HEK-293T cells with plasmids transfection. This result was further confirmed by using a FLAG-TAK1 stably expressed HeLa cell line. Radiation-induced IKKs and p38 phosphorylation were greatly impaired in MEFs reconstituted with TAK1 K158R mutant compared to ones with wild-type TAK1. Conclusions Lys63-linked TAK1 poly-ubiquitination at Lys-58 plays a key role in the process of radiation-induced NF-κB activation. |
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