Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor alpha factor promoter.
We have recently identified lipopolysaccharide tumor-induced tumor necrosis factor alpha factor (LITAF) as a novel transcription factor controlling necrosis factor (TNF)-alpha 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)-3alpha, 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-3alpha, or a nuclear binding protein to the CTCCC motif, may play an important role in regulating LPS-dependent LITAF transcription.
Shiomi
Nobuyuki
N
Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry, Okayama, Japan.
Myokai
Fumio
F
Naruishi
Koji
K
Oyaizu
Kosuke
K
Senoo
Kyoko
K
Yamaguchi
Tomoko
T
Amar
Salomon
S
Takashiba
Shogo
S
D024506:5' Flanking Region; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008070:Lipopolysaccharides / Q000276:immunology; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
C118738:LITAF protein, human; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D009841:Oligonucleotides; D012333:RNA, Messenger; D014157:Transcription Factors; D004247:DNA