Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) causes severe losses in the global pig industry. In the present study, we investigated the molecular characterization of porcine interferon stimulated gene 12a (ISG12A) and confirmed its anti-PRRSV ability for the first time. We found that porcine ISG12A was localized in mitochondria and significantly decreased the number of cells in G2/S phase. Porcine ISG12A mRNA was up-regulated in cells/tissues of Tongcheng (TC) pigs and Large White (LW) pigs after PRRSV challenge. More importantly, the ectopic overexpression of ISG12A could significantly suppress PRRSV replication at 24, 36 and 48 h post challenge (hpc), which was confirmed by detecting PRRSV ORF7 mRNA with quantitative reverse transcription polymerase chain reaction (qRT-PCR) and PRRSV N protein with indirect immunofluorescence assay (IFA) in MARC-145 cells. Meanwhile, knockdown of endogenic ISG12A could obviously facilitate PRRSV replication in MARC-145 cells at 36 hpc. The results will lead to a better understanding of the interaction between host immune system and PRRSV, which may help us develop novel therapeutic tools to control PRRSV.
Highlights
Porcine reproductive and respiratory syndrome (PRRS) is one of most economically devastating viral diseases in the global pig industry [1,2,3]
The result of the multiple sequence alignment showed that porcine interferon stimulated gene 12a (ISG12A) protein contains a conserved ISG12 motif (95–169 aa), which is consistent with homologues of monkey and human (Figure 1A)
Phylogenetic tree according to DNA sequences showed that porcine ISG12A was closer to homologues of cow and sheep (Figure 1C)
Summary
Porcine reproductive and respiratory syndrome (PRRS) is one of most economically devastating viral diseases in the global pig industry [1,2,3]. The causative pathogen, porcine reproductive and respiratory syndrome virus (PRRSV), is a single-stranded positive-sense RNA virus. Interferons (IFNs) are an important family of cytokines secreted by host cells in response to different pathogens. Interferon-stimulated genes (ISGs) induced by IFNs play important roles in IFN signaling [9]. Interferon-stimulated gene 12a (ISG12A) is a recently discovered as an antiviral ISG. The promoter region of the ISG12A gene contains an IFN-stimulated response element (ISRE), interferon regulatory factor 1/2 (IRF1/IRF2) and signal transducer and activator of transcription (STAT) binding sites [18]. As a member of the ISGs, ISG12A can dramatically restrict different viral infections, such as hepatitis C virus (HCV), vesicular stomatitis virus (VSV) and Newcastle disease virus (NDV) [21,22,23]. Whether porcine ISG12A is involved in host resistance to PRRSV infection is of great importance. The results acquired from this study will contribute to further understanding of the interaction between the host and PRRSV, which may help us develop novel therapeutic tools to control PRRSV
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