title: Cloning and characterization of lipopolysaccharide-induced tumor necrosis factor alpha factor promoter.
abstract: 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.
authors.affiliation: Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry, Okayama, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shiomi
authors.forename: Fumio
authors.initials: F
authors.lastname: Myokai
authors.forename: Koji
authors.initials: K
authors.lastname: Naruishi
authors.forename: Kosuke
authors.initials: K
authors.lastname: Oyaizu
authors.forename: Kyoko
authors.initials: K
authors.lastname: Senoo
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Salomon
authors.initials: S
authors.lastname: Amar
authors.forename: Shogo
authors.initials: S
authors.lastname: Takashiba
mesh_terms: 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
chemical_list: C118738:LITAF protein, human; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D009841:Oligonucleotides; D012333:RNA, Messenger; D014157:Transcription Factors; D004247:DNA