Role of Extracellular Signal-Regulated Kinase 1/2 and Reactive Oxygen Species in Toll-Like Receptor 2-Mediated Dual-Specificity Phosphatase 4 Expression.
10.12701/yujm.2013.30.1.10
- Author:
So Yeon KIM
1
;
Suk Hwan BAEK
Author Information
1. Department of Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, Daegu, Korea. sbaek@med.yu.ac.kr
- Publication Type:Original Article
- Keywords:
Toll-like receptor;
Dual-specificity phosphatase;
ERK1/2;
Reactive oxygen species
- MeSH:
Acridines;
Animals;
Anti-Bacterial Agents;
Blotting, Western;
Butadienes;
Cell Line;
Dual-Specificity Phosphatases;
Macrophages;
Mice;
Mitogen-Activated Protein Kinase Phosphatases;
Mitogen-Activated Protein Kinases;
Nitriles;
Penicillins;
Phosphorylation;
Phosphotransferases;
Reactive Oxygen Species;
Receptors, Pattern Recognition;
Toll-Like Receptors
- From:Yeungnam University Journal of Medicine
2013;30(1):10-16
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Toll-like receptors (TLRs) are well-known pattern recognition receptors. Among the 13 TLRs, TLR2 is the most known receptor for immune response. It activates mitogen-activated protein kinases (MAPKs), which are counterbalanced by MAPK phosphatases [MKPs or dual-specificity phosphatases (DUSPs)]. However, the regulatory mechanism of DUSPs is still unclear. In this study, the effect of a TLR2 ligand (TLR2L, Pam3CSK4) on DUSP4 expression in Raw264.7 cells was demonstrated. METHODS: A Raw264.7 mouse macrophage cell line was cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 1% antibiotics (100 U/mL penicillin and 100 g/mL streptomycin) at 37degrees C in 5% CO2. TLR2L (Pam3CSK4)-mediated DUSP4 expressions were confirmed with RT-PCR and western blot analysis. In addition, the detection of reactive oxygen species (ROS) was measured with lucigenin assay. RESULTS: Pam3CSK4 induced the expression of DUSP1, 2, 4, 5 and 16. The DUSP4 expression was also increased by TLR4 and 9 agonists (lipopolysaccharide and CpG ODN, respectively). Pam3CSK4 also induced ERK1/2 phosphorylation and ROS production, and the Pam3CSK4-induced DUSP4 expression was decreased by ERK1/2 (U0126) and ROS (DPI) inhibitors. U0126 suppressed the ROS production by Pam3CSK4. CONCLUSION: Pam3CSK4-mediated DUSP4 expression is regulated by ERK1/2 and ROS. This finding suggests the physiological importance of DUSP4 in TLR2-mediated immune response.
