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首页> 美国卫生研究院文献>Oncotarget >MicroRNA-like viral small RNA from porcine reproductive and respiratory syndrome virus negatively regulates viral replication by targeting the viral nonstructural protein 2
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MicroRNA-like viral small RNA from porcine reproductive and respiratory syndrome virus negatively regulates viral replication by targeting the viral nonstructural protein 2

机译：来自猪繁殖与呼吸综合症病毒的MicroRNA样病毒小RNA通过靶向病毒非结构蛋白2负调控病毒复制

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Many viruses encode microRNAs (miRNAs) that are small non-coding single-stranded RNAs which play critical roles in virus-host interactions. Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically impactful viruses in the swine industry. The present study sought to determine whether PRRSV encodes miRNAs that could regulate PRRSV replication. Four viral small RNAs (vsRNAs) were mapped to the stem-loop structures in the ORF1a, ORF1b and GP2a regions of the PRRSV genome by bioinformatics prediction and experimental verification. Of these, the structures with the lowest minimum free energy (MFE) values predicted for PRRSV-vsRNA1 corresponded to typical stem-loop, hairpin structures. Inhibition of PRRSV-vsRNA1 function led to significant increases in viral replication. Transfection with PRRSV-vsRNA1 mimics significantly inhibited PRRSV replication in primary porcine alveolar macrophages (PAMs). The time-dependent increase in the abundance of PRRSV-vsRNA1 mirrored the gradual upregulation of PRRSV RNA expression. Knockdown of proteins associated with cellular miRNA biogenesis demonstrated that Drosha and Argonaute (Ago2) are involved in PRRSV-vsRNA1 biogenesis. Moreover, PRRSV-vsRNA1 bound specifically to the nonstructural protein 2 (NSP2)-coding sequence of PRRSV genome RNA. Collectively, the results reveal that PRRSV encodes a functional PRRSV-vsRNA1 which auto-regulates PRRSV replication by directly targeting and suppressing viral NSP2 gene expression. These findings not only provide new insights into the mechanism of the pathogenesis of PRRSV, but also explore a potential avenue for controlling PRRSV infection using viral small RNAs.
机译：许多病毒编码的微小RNA（miRNA）是小的非编码单链RNA，在病毒与宿主之间的相互作用中起着至关重要的作用。猪繁殖与呼吸综合症病毒（PRRSV）是养猪业中最具经济影响的病毒之一。本研究试图确定PRRSV是否编码可调节PRRSV复制的miRNA。通过生物信息学预测和实验验证，将四个病毒小RNA（vsRNA）定位到PRRSV基因组的ORF1a，ORF1b和GP2a区的茎环结构。其中，针对PRRSV-vsRNA1预测的最低最低自由能（MFE）值最低的结构对应于典型的茎环发夹结构。 PRRSV-vsRNA1功能的抑制导致病毒复制的显着增加。 PRRSV-vsRNA1模拟物转染可显着抑制原代猪肺泡巨噬细胞（PAM）中的PRRSV复制。 PRRSV-vsRNA1的丰度随时间的增加反映了PRRSV RNA表达的逐渐上调。敲除与细胞miRNA生物发生有关的蛋白质表明，Drosha和Argonaute（Ago2）参与PRRSV-vsRNA1生物发生。此外，PRRSV-vsRNA1特异性结合PRRSV基因组RNA的非结构蛋白2（NSP2）编码序列。总体而言，结果表明PRRSV编码功能性PRRSV-vsRNA1，该功能通过直接靶向和抑制病毒NSP2基因表达来自动调节PRRSV复制。这些发现不仅为PRRSV的发病机理提供了新的见解，而且还探索了使用病毒小RNA控制PRRSV感染的潜在途径。

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