Hong Jae-Sang, Kim Nam-Hoon, Choi Chang-Yong, Lee Jun-Seong, Na Dokyun, Chun Taehoon, Lee Young Sik
College of Life Sciences and Biotechnology, Korea University, Seoul, 136-713, Korea.
Present address: Institut de Recherches Cliniques de Montréal, Montréal, Québec, H2W1R7, Canada.
Vet Res. 2015 Apr 10;46(1):39. doi: 10.1186/s13567-015-0172-5.
Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome, which leads to serious economic losses in the pig industry worldwide. While the molecular basis of PCV2 replication and pathogenicity remains elusive, it is increasingly apparent that the microRNA (miRNA) pathway plays a key role in controlling virus-host interactions, in addition to a wide range of cellular processes. Here, we employed Solexa deep sequencing technology to determine which cellular miRNAs were differentially regulated after expression of each of three PCV2-encoded open reading frames (ORFs) in porcine kidney epithelial (PK15) cells. We identified 51 ORF1-regulated miRNAs, 74 ORF2-regulated miRNAs, and 32 ORF3-regulated miRNAs that differed in abundance compared to the control. Gene ontology analysis of the putative targets of these miRNAs identified transcriptional regulation as the most significantly enriched biological process, while KEGG pathway analysis revealed significant enrichment for several pathways including MAPK signaling, which is activated during PCV2 infection. Among the potential target genes of ORF-regulated miRNAs, two genes encoding proteins that are known to interact with PCV2-encoded proteins, zinc finger protein 265 (ZNF265) and regulator of G protein signaling 16 (RGS16), were selected for further analysis. We provide evidence that ZNF265 and RGS16 are direct targets of miR-139-5p and let-7e, respectively, which are both down-regulated by ORF2. Our data will initiate further studies to elucidate the roles of ORF-regulated cellular miRNAs in PCV2-host interactions.
猪圆环病毒2型(PCV2)是断奶后多系统消耗综合征的主要病原体,该疾病在全球养猪业中导致严重的经济损失。虽然PCV2复制和致病性的分子基础仍不清楚,但越来越明显的是,除了广泛的细胞过程外,微小RNA(miRNA)途径在控制病毒与宿主的相互作用中起着关键作用。在这里,我们采用Solexa深度测序技术来确定在猪肾上皮(PK15)细胞中表达三种PCV2编码的开放阅读框(ORF)中的每一种后,哪些细胞miRNA受到差异调节。我们鉴定出51种受ORF1调节的miRNA、74种受ORF2调节的miRNA和32种受ORF3调节的miRNA,它们与对照相比丰度不同。对这些miRNA的假定靶标的基因本体分析确定转录调控是最显著富集的生物学过程,而KEGG通路分析显示包括MAPK信号通路在内的几种通路有显著富集,该信号通路在PCV2感染期间被激活。在ORF调节的miRNA的潜在靶基因中,选择了两个已知与PCV2编码蛋白相互作用的蛋白编码基因,即锌指蛋白265(ZNF265)和G蛋白信号调节剂16(RGS16)进行进一步分析。我们提供的证据表明,ZNF265和RGS16分别是miR-139-5p和let-7e的直接靶标,它们都被ORF2下调。我们的数据将启动进一步的研究,以阐明ORF调节的细胞miRNA在PCV2与宿主相互作用中的作用。