ZBP-89, Sp1, and Nuclear Factor-κB Regulate Epithelial Neutrophil-activating Peptide-78 Gene Expression in Caco-2 Human Colonic Epithelial Cells

Andrew C. Keates, Sarah Keates, John H. Kwon, Kristen O. Arseneau, David J. Law, Longchuan Bai, Juanita L. Merchant, Timothy C. Wang, Ciarán P. Kelly

Research output: Contribution to journalArticle

47 Scopus citations

Abstract

We reported previously that human colonic epithelial cells produce the C-X-C chemokine epithelial neutrophil-activating peptide-78 (ENA-78) and that its expression is up-regulated in ulcerative colitis. The aim of this study was to investigate the transcriptional regulation of ENA-78 gene expression in Caco-2 intestinal epithelial cells. Reporter gene transfection and electrophoretic mobility shift assay studies demonstrated that cooperation between two regions of the ENA-78 promoter were required for maximal gene expression in interleukin-1β-stimulated Caco-2 cells. Binding of activated p50/p65 nuclear factor-κB to nucleotides -82 to -91 was essential for interleukin-1β-dependent gene transcription, whereas binding of constitutively expressed zinc-requiring nuclear factors to nucleotides - 125 to - 134 (site A) was required for basal gene expression. Scanning mutagenesis of site A demonstrated overlapping binding elements at this locus. One site (CTCCCCC) bound Sp1 and Sp3, and overexpression of Sp1 (but not Sp3) up-regulated basal ENA-78 transcription. Another site (CCCCTCCCCC) was found to bind the zinc finger nuclear factor ZBP-89, and overexpression of this protein significantly repressed ENA-78 reporter gene activity. This study demonstrates that ENA-78 gene expression in Caco-2 intestinal epithelial cells is subject to complex regulation involving the coordinate binding of ZBP-89, Sp1, and nuclear factor-κB to the ENA-78 promoter.

Original languageEnglish (US)
Pages (from-to)43713-43722
Number of pages10
JournalJournal of Biological Chemistry
Volume276
Issue number47
DOIs
StatePublished - Nov 23 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Keates, A. C., Keates, S., Kwon, J. H., Arseneau, K. O., Law, D. J., Bai, L., Merchant, J. L., Wang, T. C., & Kelly, C. P. (2001). ZBP-89, Sp1, and Nuclear Factor-κB Regulate Epithelial Neutrophil-activating Peptide-78 Gene Expression in Caco-2 Human Colonic Epithelial Cells. Journal of Biological Chemistry, 276(47), 43713-43722. https://doi.org/10.1074/jbc.M107838200