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A novel p38 MAPK indentified from Crassostrea hongkongensis and its involvement in host response to immune challenges

Qu, Fufa, Xiang, Zhiming, Zhang, Yang, Li, Jun, Xiao, Shu, Zhang, Yuehuan, Mao, Fan, Ma, Haitao, Yu, Ziniu
Molecular Immunology 2016 v.79 pp. 113-124
Crassostrea gigas, Staphylococcus haemolyticus, Vibrio alginolyticus, amino acids, binding sites, complementary DNA, cytoplasm, developmental stages, dose response, gene activation, gene expression, hemocytes, homeostasis, immune response, innate immunity, lipopolysaccharides, mitogen-activated protein kinase, molecular weight, oysters, pathogens, peptidoglycans, phosphorylation, phylogeny, proteins, quantitative polymerase chain reaction, reporter genes, tissues, China
p38 mitogen-activated protein kinases (MAPKs) are conserved serine/threonine-specific kinases that are activated by various extracellular stimuli and play crucial regulatory roles in immunity, development and homeostasis. However, the function of p38s in mollusks, the second most diverse group of animals, is still poorly understood. In this study, a novel molluscan p38 (designated Chp38) was cloned and characterized from the Hong Kong oyster Crassostrea hongkongensis. Its full-length cDNA encoded a putative protein of 353 amino acids with a calculated molecular weight of approximately 40.3kDa. Similar to other reported p38 family proteins, the deduced Chp38 sequence contained a conserved dual phosphorylation TGY motif and a substrate binding site of ATRW. Phylogenetic analysis revealed that Chp38 was closest to its homolog from the Pacific oyster and belonged to the mollusk cluster. Quantitative real-time PCR analysis showed that Chp38 was constitutively expressed in all examined oyster tissues and developmental stages and that its expression in hemocytes was significantly up-regulated after pathogen (Vibrio alginolyticus and Staphylococcus haemolyticus) and PAMP (lipopolysaccharide and peptidoglycan) infections. Moreover, overexpression analysis revealed that Chp38 was localized in both the cytoplasm and nucleus of HEK293T cells and that it could significantly enhance AP-1 reporter gene activation in a dose-dependent manner. Altogether, these results provide the first experimental evidence of a functional p38 in oysters and suggest its involvement in the innate immunity of C. hongkongensis.