Suzuki Shunsuke, Singhirunnusorn Pattama, Nakano Hiroyasu, Doi Takahiro, Saiki Ikuo, Sakurai Hiroaki
Division of Pathogenic Biochemistry, Institute of Natural Medicine, University of Toyama, Toyama 930-0194, Japan.
FEBS Lett. 2006 May 29;580(13):3257-62. doi: 10.1016/j.febslet.2006.04.086. Epub 2006 May 6.
Serine protease inhibitor SerpinE2 is known as a cytokine-inducible gene. Here, we investigated whether tumor necrosis factor alpha-(TNF-alpha)-induced expression of SerpinE2 is mediated by the nuclear factor-kappaB (NF-kappaB) p65 subunit. Both steady state and TNF-alpha-induced expression of SerpinE2 mRNA were abrogated in p65-/- murine embryonic fibroblasts (MEFs). Reconstitution with wild-type p65 rescued SerpinE2 mRNA expression in an IkappaB kinase beta-dependent manner. Electrophoresis mobility shift assay and ChIP assay demonstrated that p65 bound to the kappaB-like DNA sequence located at approximately -9 kbp in the SerpinE2 promoter. In addition, TNF-alpha stimulated luciferase gene expression driven by the kappaB-like element in the reconstituted MEFs, but not in p65-/- MEFs. These results indicated that activation of NF-kappaB p65 plays an important role in TNF-alpha-induced expression of SerpinE2.
丝氨酸蛋白酶抑制剂SerpinE2是一种细胞因子诱导基因。在此，我们研究了肿瘤坏死因子α（TNF-α）诱导的SerpinE2表达是否由核因子κB（NF-κB）p65亚基介导。在p65基因敲除的小鼠胚胎成纤维细胞（MEF）中，SerpinE2 mRNA的稳态表达和TNF-α诱导的表达均被消除。用野生型p65进行重组以IκB激酶β依赖性方式挽救了SerpinE2 mRNA的表达。电泳迁移率变动分析和染色质免疫沉淀分析表明，p65与位于SerpinE2启动子中约-9 kbp处的κB样DNA序列结合。此外，TNF-α刺激了重组MEF中由κB样元件驱动的荧光素酶基因表达，但在p65基因敲除的MEF中未出现这种情况。这些结果表明，NF-κB p65的激活在TNF-α诱导的SerpinE2表达中起重要作用。
