Identification of TNF-alpha-responsive NF-kappaB p65-binding element in the distal promoter of the mouse serine protease inhibitor SerpinE2.
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.
Suzuki
Shunsuke
S
Division of Pathogenic Biochemistry, Institute of Natural Medicine, University of Toyama, Toyama 930-0194, Japan.
Singhirunnusorn
Pattama
P
Nakano
Hiroyasu
H
Doi
Takahiro
T
Saiki
Ikuo
I
Sakurai
Hiroaki
H
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus; D005347:Fibroblasts / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors; D051379:Mice; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D021381:Protein Transport; D012333:RNA, Messenger / Q000378:metabolism; D020218:Response Elements / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D058258:Serpin E2; D015843:Serpins / Q000235:genetics*; D013876:Thiophenes / Q000494:pharmacology; D051996:Transcription Factor RelA / Q000378:metabolism*; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology*
D012333:RNA, Messenger; C477523:SC 514; D058258:Serpin E2; C511869:Serpine2 protein, mouse; D015843:Serpins; D013876:Thiophenes; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D051550:I-kappa B Kinase