State Key Laboratory of Agricultural Microbiology and Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
J Neurovirol. 2018 Oct;24(5):597-605. doi: 10.1007/s13365-018-0651-3. Epub 2018 Jul 9.
Long noncoding RNAs (lncRNAs) play important roles in regulating eukaryotic genome replication and gene expression in diverse biological systems. Here, we identified lncRNAs transcribed from pseudorabies virus (PRV)-infected PK-15 cells. Based on high-throughput sequencing data, we obtained 87,263,926 and 93,947,628 clean reads from mock-infected and PRV-infected PK-15 cells, respectively. Through a normalized analytic protocol, we identified three novel viral lncRNAs. According to an analysis of differential expression between the mock-infected and PRV-infected cells, 4151 host lncRNAs were significantly upregulated and 2327 host lncRNAs were significantly downregulated in the latter group. Viral lncRNAs and several host lncRNAs were verified by northern blotting and real-time PCR. The findings showed that the viral lncRNA LDI might regulate the expression of IE180, a potent transcriptional activator of viral genes. Furthermore, we characterized the expression of viral lncRNAs in a culture of infected primary chicken dorsal root ganglia (DRG). Collectively, the obtained data suggest that PRV generates lncRNAs in both epithelial cells and chick DRG neurons.
长链非编码 RNA（lncRNA）在不同的生物系统中对于调控真核生物基因组复制和基因表达起着重要作用。在此，我们鉴定了伪狂犬病病毒（PRV）感染 PK-15 细胞中转录的 lncRNA。基于高通量测序数据，我们分别从对照感染和 PRV 感染的 PK-15 细胞中获得了 87,263,926 和 93,947,628 条清洁读段。通过标准化分析方案，我们鉴定出了三个新的病毒 lncRNA。根据对照感染和 PRV 感染细胞之间的差异表达分析，在后者中 4151 个宿主 lncRNA 显著上调，2327 个宿主 lncRNA 显著下调。通过 northern blot 和实时 PCR 验证了病毒 lncRNA 和一些宿主 lncRNA。研究结果表明，病毒 lncRNA LDI 可能调控了病毒基因的强转录激活子 IE180 的表达。此外，我们还在感染的原代鸡背根神经节（DRG）的培养物中表征了病毒 lncRNA 的表达。总之，这些数据表明 PRV 在上皮细胞和鸡 DRG 神经元中均产生 lncRNA。
