Involvement of Sp1 and Sp3 in differential regulation of human NHE3 promoter activity by sodium butyrate and IFN-γ/TNF-α
Open Access
- 1 July 2007
- journal article
- Published by American Physiological Society in American Journal of Physiology-Gastrointestinal and Liver Physiology
- Vol. 293 (1), G374-G382
- https://doi.org/10.1152/ajpgi.00128.2007
Abstract
Previously, we reported that IFN-γ and TNF-α downregulate the expression of the human Na+/H+exchanger (NHE)3 gene by modulating Sp1/Sp3 transcription factors in C2BBe1 cells. It is reported that butyrate inhibits IFN-γ and TNF-α signaling pathways. In this study, we have investigated the effect of sodium butyrate (NaB) and IFN-γ/TNF-α on human NHE3 promoter activity. In transient transfection studies, NaB (5 mM) led to 10-fold stimulation of NHE3 promoter activity after incubation for 24 h. With 5′-deletion analysis, the NaB-responsive region was mapped to the NHE3 core promoter, bp −95 to + 5, which we had shown previously to confer responsiveness to IFN-γ/TNF-α. The stimulatory effect of NaB on the NHE3 promoter was reduced by 60% in the presence of IFN-γ/TNF-α. Mutually, the repressive effect of these cytokines was attenuated by NaB. Knockdown of Sp1 and Sp3 expression with small interfering RNA (siRNA) resulted in a significant resistance to NaB effects. NaB treatment showed no effect on Sp1 and Sp3 protein expression as assessed by Western blot analyses. Gel mobility shift assays with nuclear proteins from NaB-treated cells showed enhanced binding of Sp1 and Sp3 to the NHE3 promoter. The phosphatase inhibitor okadaic acid (200 nM) blocked the stimulatory effect of NaB on the NHE3 promoter. NaB effects on the NHE3 promoter were significantly attenuated by protein phosphatase (PP)1α- and PP2Aα-specific siRNA transfection. Our data suggest that the differential regulation of NHE3 gene expression by NaB and IFN-γ/TNF-α is mediated through alternative pathways that converge on Sp1/Sp3.Keywords
This publication has 43 references indexed in Scilit:
- IFN-γ and TNF-α regulate human NHE3 gene expression by modulating the Sp family transcription factors in human intestinal epithelial cell line C2BBe1American Journal of Physiology-Cell Physiology, 2006
- Transcriptional stimulation of the human NHE3 promoter activity by PMA: PKC independence and involvement of the transcription factor EGR-1Biochemical Journal, 2006
- Subcloning, localization, and expression of the rat intestinal sodium-hydrogen exchanger isoform 8American Journal of Physiology-Gastrointestinal and Liver Physiology, 2005
- Increased Sp1 phosphorylation as a mechanism of hepatocyte growth factor (HGF/SF)-induced vascular endothelial growth factor (VEGF/VPF) transcriptionJournal of Cell Science, 2003
- In Vivo Evidence for Interferon-γ-mediated Homeostatic Mechanisms in Small Intestine of the NHE3 Na+/H+ Exchanger Knockout Model of Congenital DiarrheaPublished by Elsevier BV ,2002
- Counter-regulatory effect of sodium butyrate on tumour necrosis factor-alpha (TNF-α)-induced complement C3 and factor B biosynthesis in human intestinal epithelial cellsClinical and Experimental Immunology, 1999
- Molecular Cloning, Genomic Organization, and Functional Expression of Na+/H+ Exchanger Isoform 5 (NHE5) from Human BrainPublished by Elsevier BV ,1999
- The toxin of diarrheic shellfish poisoning, okadaic acid, increases intestinal epithelial paracellular permeabilityGastroenterology, 1997
- Dephosphorylation of Sp1 by Protein Phosphatase 1 Is Involved in the Glucose-mediated Activation of the Acetyl-CoA Carboxylase GenePublished by Elsevier BV ,1996
- Characterization of a GC-rich Region Containing Sp1 Binding Site(s) as a Constitutive Responsive Element of the α2(I) Collagen Gene in Human FibroblastsPublished by Elsevier BV ,1995