Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor α factor promoter

Nobuyuki Shiomi, Fumio Myokai, Koji Naruishi, Kosuke Oyaizu, Kyoko Senoo, Tomoko Yamaguchi, Salomon Amar, Shogo Takashiba

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor α factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-α expression in the human monocytic cell line, THP-1. To characterize the human (h) LITAF promoter, we isolated a 1.2-kb DNA fragment and followed this by a screening of human genomic DNA with a hLITAF cDNA probe. A 34-bp sequence domain located from nucleotides -74 to -43 in the hLITAF promoter exhibited the highest basal reporter gene activity; however, the activity was not elevated by lipopolysaccharide (LPS) stimulation. The sequence domain included a consensus sequence for hepatocyte nuclear factor (HNF)-3α, regulating the transcription of many kinds of genes. Interestingly, the DNA sequence position between -542 and -538 in the hLITAF promoter contained the CTCCC motif, which has been reported to act as a specific binding site for hLITAF protein. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of THP-1 nuclear factors to a 22 bp probe containing the CTCCC motif. In addition, hLITAF mRNA and nuclear hLITAF protein increased significantly in the THP-1 cells immediately after LPS stimulation. These results suggest that the consensus sequence for HNF-3α, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.

Original languageEnglish
Pages (from-to)360-368
Number of pages9
JournalFEMS Immunology and Medical Microbiology
Volume47
Issue number3
DOIs
Publication statusPublished - Aug 2006

Keywords

  • Electrophoretic mobility shift assay
  • Genomic cloning
  • LPS-induced TNF-α factor
  • Promoter sequence
  • Reporter gene assay

ASJC Scopus subject areas

  • Immunology and Allergy
  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

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