Abstract
Newcastle disease virus (NDV), causative agent of Newcastle disease (ND), is one of the most devastating pathogens for poultry industry worldwide. MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression by regulating mRNA translation efficiency or mRNA abundance through binding to mRNA directly. Accumulating evidence has revealed that cellular miRNAs can also affect virus replication by controlling host-virus interaction. To identify miRNA expression profile and explore the roles of miRNA during NDV replication, in this study, small RNA deep sequencing was performed of non-inoculated DF-1 cells (chicken embryo fibroblast cell line) and JS 5/05-infected cells collected at 6 and 12 h post infection (hereafter called mock‚ NDV-6 h, and NDV-12 h groups respectively). A total of 73 miRNAs of NDV-6 h group and 64miRNAs of NDV-12 h group were significantly differentially expressed (SDE) when compared with those in mock group. Meanwhile, 50 SDE miRNAs, including 48 up- and 2 down-regulated, showed the same expression patterns in NDV-6 h and NDV-12 h groups. qRT-PCR validation of 15 selected miRNAs’ expression patterns was consistent with deep sequencing. To investigate the role of these SDE miRNAs in NDV replication, miRNA mimics and inhibitors were transfected into DF-1 cells followed by NDV infection. The results revealed that gga-miR-451 and gga-miR-199-5p promoted NDV replication while gga-miR-19b-3p and gga-miR-29a-3p inhibited NDV replication. Further function research demonstrated gga-miR-451 suppressed NDV-induced inflammatory response via targeting YWHAZ (tyrosine3-monooxygenase/tryptophan5-monooxygenase activation protein zeta). Overall, our study presented a global miRNA expression profile in DF-1 cells in response to NDV infection and verified the roles of some SDE miRNAs in NDV replication which will underpin further studies of miRNAs’ roles between the host and the virus.
Highlights
Newcastle disease (ND), a highly contagious disease caused by virulent Newcastle disease virus (NDV), is one of the most severe avian diseases and can cause significant economic loss for the global poultry industry (Alexander, 2000)
A total of 364, 396, and 413 known miRNAs were identified in mock, NDV-6 h, and NDV-12 h DF-1 cells respectively
Recent studies have revealed that host miRNAs can affect virus infection and replication and their expression levels are often deregulated by viral infections
Summary
Newcastle disease (ND), a highly contagious disease caused by virulent Newcastle disease virus (NDV), is one of the most severe avian diseases and can cause significant economic loss for the global poultry industry (Alexander, 2000). The P gene of NDV encodes the V and W proteins in addition to the P protein via RNA editing (Samson et al, 1991; Steward et al, 1993). Innate immunity acts as the first line of host defense against virus infection by recognizing NDV infection via pattern recognition receptors (PRRs) like RIG-I (retinoic-acid inducible gene I) (Kato et al, 2006; Sun et al, 2013), MDA5 (melanoma differentiation-associated) (Rue et al, 2011), and TLR3 (Tolllike receptor 3) (Cheng et al, 2014). During the constant combat between the virus and host immune system, NDV has evolved some mechanisms to antagonize host antiviral responses. The V protein of NDV can inhibit IFN-beta promoter activation by binding to MDA-5 (Andrejeva et al, 2004) and can antagonize IFN response by degrading STAT1 to suppress Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathway (Qiu et al, 2016)
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