Interferon-γ augments CARD4/NOD1 gene and protein expression through interferon regulatory factor-1 in intestinal epithelial cells

Tadakazu Hisamatsu, Manabu Suzuki, Daniel K. Podolsky

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Although intestinal epithelial cells appear to be functionally hyporesponsive to normal intestinal flora, human intestinal epithelial cells can respond to enteroinvasive bacteria and induce an inflammatory response. This initial inflammatory response leads to the recruitment of polymorphonuclear leukocytes to the affected site in vitro and in vivo. CARD4/NOD1 is a potential cytosolic receptor for peptidoglycan in mammalian cells that resembles pathogen-resistant proteins of plants. In this context, CARD4/NOD1 is a candidate for a recognition protein of intracellular bacteria or peptidoglycan in intestinal epithelial cells. In this study, we demonstrate that CARD4/NOD1 is constitutively expressed in intestinal epithelial cell lines and isolated primary intestinal epithelial cells. Interferon-γ (IFNγ), which is a potent pro-inflammatory cytokine in intestinal mucosal inflammation, activates CARD4/NOD1 mRNA transcription in a time- and dose-dependent manner and results in augmentation of CARD4/NOD1 protein in SW480 cells. Promoter analysis of CARD4/NOD1 indicates that interferon regulatory factor-1 (IRF-1) binding motif (-791 to -782) is essential for the effect of IFNγ. Nuclear extracts from SW480 cells treated with IFNγ show specific binding of oligonucleotides corresponding to this IRF-1-binding motif, which was supershifted by anti-IRF-1 antibody in electrophoretic mobility shift assay. Overexpression of IRF-1 protein activates the CARD4/NOD1 promoter but not the deletion mutant of the IRF-1-binding site in a co-transfection assay of IRF-1 expression plasmid with CARD4/NOD1 promoter. These studies suggest that the Th1 cytokine, IFNγ, activates CARD4/NOD1 transcription and regulate innate immune mechanisms in the condition of intestinal mucosal inflammation.

Original languageEnglish
Pages (from-to)32962-32968
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number35
DOIs
Publication statusPublished - 2003 Aug 29

Fingerprint

Interferon Regulatory Factor-1
Interferons
Epithelial Cells
Nod1 Signaling Adaptor Protein
Gene Expression
Proteins
Transcription
Assays
Bacteria
Cytokines
Inflammation
Electrophoretic mobility
Plant Proteins
Peptidoglycan
Electrophoretic Mobility Shift Assay
Pathogens
Cell Extracts
Oligonucleotides
Transfection
Neutrophils

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interferon-γ augments CARD4/NOD1 gene and protein expression through interferon regulatory factor-1 in intestinal epithelial cells. / Hisamatsu, Tadakazu; Suzuki, Manabu; Podolsky, Daniel K.

In: Journal of Biological Chemistry, Vol. 278, No. 35, 29.08.2003, p. 32962-32968.

Research output: Contribution to journalArticle

Hisamatsu, Tadakazu ; Suzuki, Manabu ; Podolsky, Daniel K. / Interferon-γ augments CARD4/NOD1 gene and protein expression through interferon regulatory factor-1 in intestinal epithelial cells. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 35. pp. 32962-32968.
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