LGP2 of black carp plays an important role in the innate immune response against SVCV and GCRV

Abstract

RIG-I like receptors (RLRs) detect cytosolic RNA virus components and initiate antiviral innate immune response through downstream signaling in vertebrates. In this paper, LGP2 of black carp (Mylopharyngodon piceus) has been cloned and characterized, which is a key member of RLR family. The full-length cDNA of black carp LGP2 (bcLGP2) comprises 2941 nucleotides and the predicted bcLGP2 protein contains 682 amino acids. bcLGP2 shares core homologous structural domains of RLRs, including a N-terminnal DExD/H helicase domain, a helicase superfamily c-terminal domain, and a C-terminal regulatory domain (CTD). bcLGP2 mRNA was constitutively detected in all selected tissues including heart, liver, spleen, kidney, intestine, muscle, skin, gill; and bcLGP2 mRNA level was increased in all the tissues except gill in response to GCRV or SVCV infection. Q-PCR of Mylopharyngodon piceus fin (MPF) cells demonstrated that bcLGP2 transcription was up-regulated by Poly (I:C) treatment, GCRV or SVCV infection, but not by LPS or PMA treatment. Western blot analysis demonstrated that the molecular weight of bcLGP2 was around 80 KDa; and the immunofluorescence staining of both HeLa cells and EPC cells showed that bcLGP2 was a cytosolic protein. EPC cells transfected with plasmid expressing bcLGP2 showed obviously improved antiviral ability against SVCV and GCRV. In general, these data support the conclusion that bcLGP2 functions importantly in the host antiviral innate immune response.