Molecular cloning and expression of dsDNA cytosolic sensor gene, DDX41 from olive flounder (Paralichthys olivaceus)
트루옹 퀴 누
Molecular cloning and expression of dsDNA cytosolic sensor gene DDX41 from olive flounder (Paralichthys olivaceus);
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Abstract Pathogenic DNA recognition receptors such as the DExD family play an important role in the immune innate response among organisms. In mammals, DDX41, one of the receptors belonging to the DExD family of helicases, has been known to recognize microbial nucleic acids in the cytoplasm and enhances antiviral response in host cells. However, its mechanism and structure in teleosts remain poorly understood. Here, we cloned and sequenced the full length cDNA of DDX41 in Japanese flounder (JfDDX41) and investigated its role in immune response to viral infection and sensing bacterial cyclic dinucleotides. JfDDX41 consists of 2269 nucleotides with 155 bp 5'-UTR and 269 bp 3'- UTR encoding 614 amino acids residues. The JfDDX41 gene was subsequently determined, and it was revealed to be 5812 bp in length and includes 17 exons and 16 introns. At the amino acid sequence level, the full-length JfDDX41 showed high homology with the DDX41s of other vertebrates: 94% with medaka, 92% with tetraodon and fugu, 91% with stickleback and 76% with mammals (i.e. mouse, and human). JfDDX41 mRNA is located in mDCs and is distinctly expressed in fish naturally infected with LCDV. High expression levels were examined in gill, liver, kidney and skin; whereas eyes showed the lowest expression. The JfDDX41 can detect and bind with C-di-GMP, which lead to the activation of innate immune response. Reporter assay results showed that mRNA expression levels of type I IFN were increased in the HINAE cells treated with JfDDX41. The results suggest that JfDDX41 plays an important role in the recognition of cytoplasmic DNA to induce antiviral activity by the production of IFN and regulate proinflammatory moleculestimulated proteins.