小鼠丝氨酸蛋白酶抑制剂SerpinE2远端启动子中肿瘤坏死因子-α反应性核因子-κB p65结合元件的鉴定
Identification of TNF-alpha-responsive NF-kappaB p65-binding element in the distal promoter of the mouse serine protease inhibitor SerpinE2.
作者信息
Suzuki Shunsuke, Singhirunnusorn Pattama, Nakano Hiroyasu, Doi Takahiro, Saiki Ikuo, Sakurai Hiroaki
机构信息
Division of Pathogenic Biochemistry, Institute of Natural Medicine, University of Toyama, Toyama 930-0194, Japan.
出版信息
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。
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