Human macrophage inflammatory protein-3alpha/CCL20/LARC/Exodus/SCYA20 is transcriptionally upregulated by tumor necrosis factor-alpha via a non-standard NF-kappaB site.
The 5'-flanking sequences of the human macrophage inflammatory protein-3alpha/CCL20 gene were cloned and transfected into G-361 human melanoma cells in a luciferase reporter construct. Tumor necrosis factor-alpha (TNF-alpha) treatment stimulated luciferase expression, and promoter truncations demonstrated that TNF-alpha inducibility is conferred by a region between nt -111 and -77, which contains a non-standard nuclear factor-kappaB (NF-kappaB) binding site. The requirement for NF-kappaB was demonstrated as follows: (i) mutations in this NF-kappaB site abrogated TNF-alpha responsiveness; (ii) TNF-alpha activated a construct containing two copies of the CCL20 NF-kappaB binding site; (iii) overexpression of NF-kappaB p65 activated the CCL20 promoter; (iv) NF-kappaB from nuclear extracts of TNF-alpha-stimulated cells bound specifically to this NF-kappaB site.
Harant
H
H
Department of Inflammatory Diseases, Novartis Research Institute, Vienna, Austria.
Eldershaw
S A
SA
Lindley
I J
IJ
D024506:5' Flanking Region / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D054418:Chemokine CCL20; D019742:Chemokines, CC / Q000235:genetics*; D003001:Cloning, Molecular; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D054399:Receptors, CCR6; D019707:Receptors, Chemokine; D020218:Response Elements / Q000235:genetics*; D051996:Transcription Factor RelA; D014158:Transcription, Genetic / Q000187:drug effects*; D015533:Transcriptional Activation / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
C108355:CCL20 protein, human; C106311:CCR6 protein, human; D054418:Chemokine CCL20; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D016328:NF-kappa B; D054399:Receptors, CCR6; D019707:Receptors, Chemokine; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases