| 引用本文： |
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白红妹,陈小平,陈正虎,等.TRAF2 K48聚泛素化位点的研究[J].同济大学学报（医学版）,2014,35(5):26-32. [点击复制]
- BAI Hong-mei,CHEN Xiao-ping,CHEN Zheng-hu,et al.Identification of the sites for K48-polyubiquitination regulation of human TRAF2 activities[J].同济大学学报（医学版）,2014,35(5):26-32. [点击复制]
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| 摘要: |
| 目的筛选K48聚泛素链在肿瘤坏死因子受体相关因子2（tumor necrosis factor receptor-associated facto r 2,T RA F2）上的主要修饰位点。方法比较不同物种间的T RA F2氨基酸序列,选出人T RA F2上保守率在90%以上的赖氨酸。构建人野生型TRAF2的表达载体及保守赖氨酸突变为精氨酸的突变表达载体。将不同的TRAF2表达载体与NF-κB荧光素酶报告基因表达载体共转至293FT细胞中,通过荧光素酶检测NF-κB激活情况。结果人TRAF2上共有8个保守率在90%以上的赖氨酸,酶切及测序结果证明本研究成功构建TRAF2野生型和突变型表达载体。NF-κB荧光素酶报告基因检测证明TRAF2-K320可能是K48聚泛素链的主要修饰位点。结论 TRAF2-K320位点对TRAF2介导的NF-κB激活具有负向调控作用。 |
| 关键词: 肿瘤坏死因子受体相关因子2 K48聚泛素化 赖氨酸 NF-κB |
| DOI：10.3969/j. issnl008 -0392.2014.05.006 |
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| 基金项目:国家自然科学基金（31170824） |
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| Identification of the sites for K48-polyubiquitination regulation of human TRAF2 activities |
| BAI Hong-mei,CHEN Xiao-ping,CHEN Zheng-hu,YANG Jian-hua |
| (Dept. of Immunology, School of Medicine, Tongji University, Shanghai 200092, China) |
| Abstract: |
| Objective To identify the sites involved in the regulation of human tumor necrosis factor receptor-associated factor 2（ TRA F2） activities by the K48-poly ubiquitination.Methods The conserved lysine residues on hum an TRA F2 were sorted by aligningthe hum an TRA F2 am inoacid sequences with TRA F2 from other organism s. The wild ty pe TRA F2 and sitedirected m utations of TRA F2 in which conserved ly sine was changed intoarginine were cloned intopc D N A3. 1（ ＋）. A fter co-transfectingthe ex pression vectors encodingwild ty pe TRA F2 or m utants alongwith N F-κB luciferase reporter constructs into293 FTcells,the luciferase activities were determ ined toidentify the ly sine which is involved in downregulation of TRA F2-m ediated N F-κB activation. Results Eight conserved ly sine residues were found in hum an TRA F2.Restriction enzy m e digestion analy sis and D N A sequencingdem onstrate that wild ty pe TRA F2 and m utants were constructed successfully. The result of luciferase reporter gene assay suggested that TRA F2-K320 was a potential ubiquitin acceptor site tomediate TRAF2 K48-polyubiquitination.Conclusion K48-polyubiquitination of human TRAF2 at lysine 320 may play a negative regulatory role in TRAF2-mediated NF-κB activation. |
| Key words: TRAF2 K48-polyubiquitination lysine NF-κB |
