Mx1 of black carp functions importantly in the antiviral innate immune response

Abstract

Mx (myxovirus resistance) is an important antiviral protein in the innate immune responses of vertebrates to microbial pathogens. In this study, we cloned and characterized Mx1 of black carp (Mylopharyngodon piceus). The full-length cDNA of black carp Mx1 (bcMx1) consists of 2781 nucleotides and the predicted bcMx1 protein contains 631 amino acids. bcMx1 contains a GTPase domain at the N-terminnus, a “central interactive domain” in the middle and a GTPase effector domain at the C-terminus. bcMx1 mRNA was constitutively transcribed in all tissues tested, including the heart, liver, spleen, kidney, intestine, muscle, skin and gill; and bcMx1 mRNA levels increased in all but the gill after grass carp reovirus (GCRV) or viraemia of carp virus (SVCV) infection. Quantitative PCR analysis of Mylopharyngodon piceus fin (MPF) cells indicated that bcMx1 mRNA levels increased after GCRV or SVCV infection at different multiplicities of infection (MOI). Western blotting demonstrated that the molecular weight of bcMx1 is ∼75 kDa and immunofluorescent staining data of both HeLa cells and EPC cells showed that bcMx1 is a cytosolic protein. EPC cells transfected with plasmid expressing bcMx1 showed increased antiviral activity against SVCV and GCRV. All our data suggest that bcMx1 is an antiviral protein in the innate immune response of the black carp.