ESE-1, an enterocyte-specific Ets transcription factor, regulates MIP-3alpha gene expression in Caco-2 human colonic epithelial cells.
We have previously shown that colonic epithelial cells are a major site of MIP-3alpha production in human colon and that enterocyte MIP-3alpha protein levels are elevated in inflammatory bowel disease. The aim of this study was to determine the molecular mechanisms regulating MIP-3alpha gene transcription in Caco-2 intestinal epithelial cells. ... We show that a kappaB element at nucleotides -82 to -93 of the MIP-3alpha promoter binds p50/p65 NF-kappaB heterodimers and is a major regulator of basal and interleukin-1beta (IL-1beta)-mediated gene activation. Scanning mutagenesis of the MIP-3alpha 5'-flanking region also identified two additional binding elements: Site X (nucleotides -63 to -69) and Site Y (nucleotides -143 to -154). Site X (CGCCTTC) bound Sp1 and regulated basal MIP-3alpha gene transcription. Overexpression of Sp1 increased basal luciferase activity, whereas, substitutions in the Sp1 element significantly reduced reporter activity. In contrast, Site Y (AAGCAGGAAGTT) regulated both basal and cytokine-induced gene activation and bound the Ets nuclear factor ESE-1. Substitutions in the Site Y element markedly reduced inducible MIP-3alpha reporter activity. Conversely, overexpression of ESE-1 significantly up-regulated MIP-3alpha luciferase levels. Taken together, our findings demonstrate that co-ordinate activation and binding of ESE-1, Sp1, and NF-kappaB to the MIP-3alpha promoter is required for maximal gene expression by cytokine-stimulated Caco-2 human intestinal epithelial cells.
Mesh Terms:
Base Sequence, Binding Sites, CCAAT-Enhancer-Binding Proteins, Caco-2 Cells, Chemokine CCL20, Chemokines, CC, Cloning, Molecular, DNA-Binding Proteins, Gene Expression Regulation, Humans, Interleukin-1, Macrophage Inflammatory Proteins, Molecular Sequence Data, NF-kappa B, Promoter Regions, Genetic, Proto-Oncogene Proteins, Receptors, CCR6, Receptors, Chemokine, Sp1 Transcription Factor, Sp3 Transcription Factor, Trans-Activators, Transcription Factors, Transcription, Genetic, Transcriptional Activation
Base Sequence, Binding Sites, CCAAT-Enhancer-Binding Proteins, Caco-2 Cells, Chemokine CCL20, Chemokines, CC, Cloning, Molecular, DNA-Binding Proteins, Gene Expression Regulation, Humans, Interleukin-1, Macrophage Inflammatory Proteins, Molecular Sequence Data, NF-kappa B, Promoter Regions, Genetic, Proto-Oncogene Proteins, Receptors, CCR6, Receptors, Chemokine, Sp1 Transcription Factor, Sp3 Transcription Factor, Trans-Activators, Transcription Factors, Transcription, Genetic, Transcriptional Activation
J. Biol. Chem.
Date: Jan. 10, 2003
PubMed ID: 12414801
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