Trim6结合并降解接头蛋白TAB2抑制NF-κB活化

The binding and degradation of adaptor proteins TAB2 by Trim6 inhibit TAB2-induced NF-κB activation

观察Trim6是否与接头蛋白TAB2存在蛋白相互作用,进而影响TAB2介导的NF-κB荧光素酶报告基因的活化。在HEK293T细胞中共转染Trim6及TAB2的真核表达载体,免疫共沉淀法验证两者是否存在蛋白相互作用;用蛋白印迹法验证Trim6是否能降解TAB2;同时用双荧光素酶报告基因系统检测Trim6是否影响TAB2介导的NF-κB荧光素酶报告基因活化。免疫共沉淀结果证明Trim6与TAB2存在蛋白相互作用;蛋白印迹检测发现Trim6能显著降解TAB2,进而明显抑制TAB2介导的NFκB荧光素酶报告基因的活化。

To investigate the protein interaction between Trim6 and TAB2, moreover to identify the effects of Trim6 on NF-κB luciferase reporter gene activation induced by TAB2, the interaction between Trim6 and TAB2 was examined by coimmunopre cipitation. Then, whether Trim6 could target TAB2 for degradation was detected by Western blot. Furthermore, the effects of Trim6 on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Coimmunopreeipitation ex periments indicated that Trim6 interacted with TAB2. Besides, Trim6 could target TAB2 for degradation. As a result, dual-lu- ciferase assay showed that Trim6 could effectively block TAB2 induced NF-κB activation. The result of this study suggests that Trim6 was involved in negative regulation of TAB2-induced NF-κB activation though interacting with, and degrading TAB2.