Zoological Research ›› 2019, Vol. 40 ›› Issue (2): 77-88.doi: 10.24272/j.issn.2095-8137.2018.066

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Involvement of ayu NOD2 in NF-κB and MAPK signaling pathways: Insights into functional conservation of NOD2 in antibacterial innate immunity

Yi Ren1,#, Shui-Fang Liu1,#, Li Nie1,*, Shi-Yu Cai1, Jiong Chen1,2,*   

  1. 1 Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo Zhejiang 315211, China
    2 Key Laboratory of Applied Marine Biotechnology of Ministry of Education, Ningbo University, Ningbo Zhejiang 315211, China
  • Online:2019-03-18 Published:2019-01-11
  • Contact: Li Nie, Jiong Chen, E-mail:nieli@nbu.edu.cn; jchen1975@163.com
  • Supported by:
     

Abstract: Nucleotide oligomerization domain 2 (NOD2) is a major cytoplasmic sensor for pathogens and is critical for the clearance of cytosolic bacteria in mammals. However, studies regarding NOD2, especially the initiated signaling pathways, are scarce in teleost species. In this study, we identified a NOD2 molecule (PaNOD2) from ayu (Plecoglossus altivelis). Bioinformatics analysis showed the structure of NOD2 to be highly conserved during vertebrate evolution. Dual-luciferase reporter assays examined the activation of NF-κB signaling and Western blotting analysis detected the phosphorylation of three MAP kinases (p-38, Erk1/2, and JNK1/2). Functional study revealed that, like its mammalian counterparts, PaNOD2 was the receptor of the bacterial cell wall component muramyl dipeptide (MDP), and the leucine-rich repeat motif was responsible for the recognition and binding of PaNOD2 with the ligand. Overexpression of PaNOD2 activated the NF-κB signaling pathway, leading to the upregulation of inflammatory cytokines, including TNF-α and IL-1β in HEK293T cells and ayu head kidney-derived monocytes/macrophages (MO/MΦ). Particularly, we found that PaNOD2 activated the MAPK signaling pathways, as indicated by the increased phosphorylation of p-38, Erk1/2, and JNK1/2, which have not been characterized in any teleost species previously. Our findings proved that the NOD2 molecule and initiated pathways are conserved between mammals and ayu. Therefore, ayu could be used as an animal model to investigate NOD2-based diseases and therapeutic applications.

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