Transcriptional regulation of beta-defensin-2 by lipopolysaccharide in cultured human cervical carcinoma (HeLa) cells.
Human beta-defensin-2 (hBD-2) is an antimicrobial peptide with a broad spectrum of antimicrobial activity against bacteria, yeast and fungi. Here, we analyzed the transcriptional regulation of hBD-2 in cultured human cervical carcinoma (HeLa) cells with or without lipopolysaccharide (LPS). DNA from position -329 to -39 in the hBD-2 promoter region contained the consensus binding sites for transcription factors, one site for nuclear factor for IL-6 expression (NF-IL6) and two sites for nuclear factor-(kappa)B (NF-(kappa)B). Reporter gene assays for promoter activity revealed that the region had the highest level of responsiveness to LPS. Furthermore, mutations in both of the NF-(kappa)B binding sites caused a significant reduction of the responsiveness to LPS, whereas mutation in the NF-IL6 binding site resulted in an elevation of the basal promoter activity. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of HeLa nuclear factors to 60-bp probe containing the two NF-(kappa)B binding sites, suggesting that the sites were essential for the binding. Our results suggest that the two NF-(kappa)B binding sites contribute to LPS-mediated hBD-2 transcription while the NF-IL6 binding site represses LPS-independent hBD-2 transcription in the HeLa cells.
Mineshiba
Junji
J
Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8525, Japan.
Myokai
Fumio
F
Mineshiba
Fumi
F
Matsuura
Kaori
K
Nishimura
Fusanori
F
Takashiba
Shogo
S
D001483:Base Sequence; D001665:Binding Sites; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000276:immunology; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008070:Lipopolysaccharides / Q000276:immunology*; D008969:Molecular Sequence Data; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014158:Transcription, Genetic; D023083:beta-Defensins / Q000235:genetics*
D022782:CCAAT-Enhancer-Binding Protein-beta; C106641:DEFB4A protein, human; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009841:Oligonucleotides; D023083:beta-Defensins