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Identification of TNF‐α‐responsive NF‐κB p65‐binding element in the distal promoter of the mouse serine protease inhibitor SerpinE2
Author(s) -
Suzuki Shunsuke,
Singhirunnusorn Pattama,
Nakano Hiroyasu,
Doi Takahiro,
Saiki Ikuo,
Sakurai Hiroaki
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.04.086
Subject(s) - microbiology and biotechnology , electrophoretic mobility shift assay , tumor necrosis factor alpha , luciferase , serine protease , gene expression , protein subunit , serine , biology , messenger rna , gene , chemistry , protease , transfection , enzyme , phosphorylation , biochemistry , immunology
Serine protease inhibitor SerpinE2 is known as a cytokine‐inducible gene. Here, we investigated whether tumor necrosis factor α‐(TNF‐α)‐induced expression of SerpinE2 is mediated by the nuclear factor‐κB (NF‐κB) p65 subunit. Both steady state and TNF‐α‐induced expression of SerpinE2 mRNA were abrogated in p65 −/− murine embryonic fibroblasts (MEFs). Reconstitution with wild‐type p65 rescued SerpinE2 mRNA expression in an IκB kinase β‐dependent manner. Electrophoresis mobility shift assay and ChIP assay demonstrated that p65 bound to the κB‐like DNA sequence located at approximately −9 kbp in the SerpinE2 promoter. In addition, TNF‐α stimulated luciferase gene expression driven by the κB‐like element in the reconstituted MEFs, but not in p65 −/− MEFs. These results indicated that activation of NF‐κB p65 plays an important role in TNF‐α‐induced expression of SerpinE2 .