Cited by 81Immune Response and InflammationNF-κB Signaling PathwaysT-cell and B-cell Immunology

MEK Is a Key Modulator for TLR5-induced Interleukin-8 and MIP3α Gene Expression in Non-transformed Human Colonic Epithelial Cells

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Rhee, Sang Hoon, Keates, Andrew C., Moyer, Mary P., Pothoulakis, Charalabos·2004·Journal of Biological Chemistry

Abstract

Flagellin, a specific ligand for Toll-like receptor 5 (TLR5), is a molecular pattern associated with several bacterial species. Recently, TLR signaling has been intensively studied. However, TLR5-associated signaling in non-transformed colonocytes has not been investigated. Here we studied the expression of cytokines induced by flagellin in non-transformed human colonic NCM460 cells and the signaling mechanisms mediating these responses. Cytokine expression array experiments showed that exposure of the cells to flagellin (100 ng/ml) for 12 h increased the expression of interleukin (IL)-8 and macrophage-inflammatory protein 3alpha (MIP3alpha) in a TLR5-specific manner. Flagellin also activated MAP kinases (ERK1/2, JNK, and p38) and degraded IkappaBalpha. Dominant negative MEK1 (a kinase that activates ERK1/2) blocked flagellin-stimulated IL-8 and MIP3alpha transcriptional activity, while the MEK-specific inhibitors PD98059 and U0126 reduced protein production of these cytokines. Conversely, transfection with a constitutively active MEK1 increased IL-8 and MIP3alpha transcriptional activity in a NFkappaB-independent manner. Furthermore, overexpression of the constitutively active MEK1 induced IL-8 and MIP3alpha protein production. We also demonstrated that C-terminal coiled-coil and TRAF-C domains of TRAF6, unable to mediate NFkappaB activation, are involved in MEK-mediated IL-8 and MIP3alpha expression. Thus, in non-transformed human colonocytes, MEK activation following flagellin/TLR5 engagement is a key modulator for NFkappaB-independent, IL-8 and MIP3alpha expression.

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Crossref · 10.1074/jbc.m400967200

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  • Signals that originate from luminal micro-organisms and influence the gut epithelium involve well-characterized mechanisms, such as Toll-like receptor signaling and signaling by bipeptides or tripeptides.

    “Signals that originate from luminal micro-organisms and influence the gut epithelium have been studied extensively, and involve well-characterized mechanisms, such as Toll-like receptor signaling48,49 and signaling by bipeptides or tripeptides, such as N-formylmethionyl–leucyl–phenylalanine.50”

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Authors
Rhee, S. H., Keates, A. C., Moyer, M. P., & Pothoulakis, C.
Affiliations
Beth Israel Deaconess Medical Center (US)
Corresponding author
Pothoulakis, Charalabos
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2004-06
Volume
279
Issue
24
Pages
25179-25188
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2011944491
Cited by
81
References
42
Citations by year
2023: 1 · 2020: 2 · 2019: 4 · 2017: 5 · 2016: 2 · 2015: 3 · 2014: 4 · 2013: 2 · 2012: 4
FWCI
3.24
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Institute of Diabetes and Digestive and Kidney Diseases (P01DK033506, DK-33506, DK 47-343, R01DK047343, DK-60729, R01DK060729)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
MEK1, TLR5, flagellin, interleukin-8, colonic epithelial cells, NCM460 cells, MAPK signaling, ERK1/2, TRAF6, NF-κB-independent signaling, cytokine gene expression, IκBα degradation
MeSH terms
Colon, Gene Expression Regulation, Humans, Intestinal Mucosa, Membrane Glycoproteins, Proteins, Receptors, Cell Surface, Signal Transduction, Interleukin-8, NF-kappa B, Macrophage Inflammatory Proteins, Chemokines, CC, Mitogen-Activated Protein Kinases, Mitogen-Activated Protein Kinase Kinases, TNF Receptor-Associated Factor 6, MAP Kinase Kinase 1, Toll-Like Receptors, Toll-Like Receptor 5, Chemokine CCL20

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