Cloning and characterisation of p38 MAP kinase from Atlantic salmon A kinase important for regulating salmon TNF-2 and IL-1beta expression.
p38 mitogen-activated protein kinase is activated by environmental stress and cytokines and plays a role in transcriptional regulation and inflammatory responses. In this study, three distinct Atlantic salmon p38 (As-p38) cDNAs were cloned, which all translated into 361 amino acid proteins. The As-p38 protein sequences possessed showed >85% identity to the mammalian homolog, p38alpha. All three contained the conserved phosphorylation motif TGY located in the activation loop of the kinase. Salmon p38 showed ubiquitous tissue distribution, including expression in the immune organs head kidney and spleen. A higher p38 mRNA expression was detected in the ovary compared to other organs suggesting that p38 may perform specific functions within this organ. Western blot analysis with an antibody specific for phosphorylated p38 showed that ectopically expressed As-p38 variants were activated in CHSE-214 cells in response to chemical stress. Furthermore, lipopolysaccharide, CpG oligonucleotides and recombinant trout IL-1beta induced endogenous phosphorylation of p38 in salmon head kidney macrophages in a dose-dependent manner. The importance of p38 for regulation of salmon innate immunity was further demonstrated by the ability of the p38 specific inhibitor SB203580 to completely abolish LPS-stimulated TNF-2 and IL-1beta mRNA expression in the macrophages.
Hansen
Tom E
TE
The Norwegian Structural Biology Centre and Department of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, N-9037 Tromsø, Norway.
Jørgensen
Jorunn B
JB
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D053583:Interleukin-1beta / Q000235:genetics*; D008264:Macrophages / Q000378:metabolism; D010053:Ovary; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D020317:Salmo salar / Q000276:immunology*; D014018:Tissue Distribution; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000502:physiology*
D018076:DNA, Complementary; D053583:Interleukin-1beta; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases