PMID: 15223809Jun 30, 2004Paper

A role for NF-kappa B binding motifs in the differential induction of chemokine gene expression in human corneal epithelial cells

Investigative Ophthalmology & Visual Science
Mary H RitchieJohn E Oakes

Abstract

The interleukin (IL)-8 promoter possesses a NF-kappa B-binding site with affinity to p50p65 and p65p65 complexes while the monocyte chemoattractant protein (MCP)-1 promoter's NF-kappa B-binding site has exclusive affinity to p50p65 heterodimers. The purpose of this study was to determine whether the two NF-kappa B sites play a role in the capacity of tumor necrosis factor (TNF)-alpha-stimulated human corneal epithelial cells (HCECs) to produce nanogram amounts of IL-8 in the absence of MCP-1 synthesis. IL-8 and MCP-1 promoters were cloned into luciferase reporter vectors. Site-directed mutagenesis of wild-type promoters was used to mutate the NF-kappa B-binding motif in the wild-type IL-8 reporter plasmid into a motif with exclusive affinity to p50p65 and to mutate the NF-kappa B binding motif in the wild-type MCP-1 reporter plasmid into a motif with affinity to p65p65. Luciferase activity was determined after transfection of reporter vector constructs into TNF-alpha-stimulated HCECs. The chromatin immunoprecipitation assay was used to confirm binding of NF-kappa B subunits to IL-8 and MCP-1 promoters in vivo. Promoters with affinity to p65p65 homodimers were active in driving the expression of the reporter gene, whereas promot...Continue Reading

Citations

Aug 26, 2009·BMC Molecular Biology·Mervi RistolaHarry Holthöfer
Jan 27, 2007·Experimental Eye Research·Gabriel SosneMichelle Kurpakus Wheater
Jun 12, 2010·Annals of the New York Academy of Sciences·Gabriel SosneHoward Matthew
Jul 6, 2005·International Journal of Obesity : Journal of the International Association for the Study of Obesity·J N Fain, A K Madan
May 15, 2007·Annals of the New York Academy of Sciences·Gabriel SosneMichelle Kurpakus-Wheater
Jun 8, 2006·Neoplasia : an International Journal for Oncology Research·Yuet-Kin LeungShuk-Mei Ho

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