Molecular characterization and transcriptional conservation of N-myc-interactor, Nmi, by type I and type II IFNs in mandarin fish Siniperca chuatsi

Abstract

N-myc-interactor (Nmi) belongs to interferon (IFN) stimulated genes (ISGs) and is involved in the regulation of physiological processes including viral infection, inflammatory response, apoptosis and tumorigenesis in mammals. However, the function of Nmi in teleost fish remains to be explored. In this study, an Nmi homologue was characterized from mandarin fish Siniperca chuatsi. The mandarin fish Nmi shares two conserved functional Nmi/IFP35 homology domains (NIDs) with mammalian Nmi protein in its C-terminal domain and a coiled coil region (CC) in its N-terminal domain, with its genomic DNA sequence consisting of nine exons and eight introns. Subcellular localization analysis shows that mandarin fish Nmi is a cytoplasmic protein and that its localization is dependent on the CC and NID1 regions. High and constitutive mRNA level of Nmi was observed in all examined tissues, with the highest level being observed in blood. In addition, the Nmi gene was significantly induced in various organs/tissues following the infection of infectious spleen and kidney necrosis virus (ISKNV), and its mRNA and protein level was also significantly induced in vitro after the treatment of IFNh, IFNc, as well as IFN-γ. The dual luciferase activity analysis indicated that the Nmi promoter was activated by the three type I IFNs through interferon-stimulated response element (ISRE) sites, and it can be also transcriptionally activated by IFN-γ via IRF1 which can activate the expression of Nmi through ISRE. Taken together, it is demonstrated in this study that the transcription of Nmi in mandarin fish can be regulated by type I and type II IFNs, thus confirming that Nmi in fish is also an ISG, and is involved in antiviral and IFN-induced innate immunity.

Keywords: IFN; Immune response; Mandarin fish; N-Myc-interactor; Nmi.