Expression of pseudorabies virus-encoded long noncoding RNAs in epithelial cells and neurons
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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.
KeywordslncRNA Pseudorabies virus Epithelial cell Chick DRG neuron
We thank Bin Wu (Huazhong Agricultural University) for the generous gift of the PRV SMX new variant strain. We also thank Liwen Bianji, Edanz Group China (www.liwenbianji.cn/ac), for editing the English text of a draft of this manuscript.
This work was supported by the Natural Science Foundation of China (31770191, 31470259) and National Key Research and Development Program (2016YFD0500105) to Z.F. Liu.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- Ben-Porat T, Kaplan AS (1985) Molecular biology of pseudorabiesvirus. Plenum Press, New YorkGoogle Scholar
- Iyer MK, Niknafs YS, Malik R, Singhal U, Sahu A, Hosono Y, Barrette TR, Prensner JR, Evans JR, Zhao S, Poliakov A, Cao X, Dhanasekaran SM, Wu YM, Robinson DR, Beer DG, Feng FY, Iyer HK, Chinnaiyan AM (2015) The landscape of long noncoding RNAs in the human transcriptome. Nat Genet 47:199–208CrossRefGoogle Scholar
- Li C, Fitzgerald ME, Del Mar N, Cuthbertson-Coates S, LeDoux MS, Gong S, Ryan JP, Reiner A (2015) The identification and neurochemical characterization of central neurons that target parasympathetic preganglionic neurons involved in the regulation of choroidal blood flow in the rat eye using pseudorabies virus, immunolabeling and conventional pathway tracing methods. Front Neuroanat 9:65PubMedPubMedCentralGoogle Scholar
- Li Y, Zheng G, Y Zhang, Yang X, Liu H, Chang H, Wang X, Zhao J, Wang C, Chen L (2017). MicroRNA analysis in mouse neuro-2a cells after pseudorabies virus infection. J NeuroVirol 1–11Google Scholar
- Mahjoub N, Dhorne-Pollet S, Fuchs W, Endale Ahanda ML, Lange E, Klupp B, Arya A, Loveland JE, Lefevre F, Mettenleiter TC, Giuffra E (2015) A 2.5-kilobase deletion containing a cluster of nine microRNAs in the latency-associated-transcript locus of the pseudorabies virus affects the host response of porcine trigeminal ganglia during established latency. J Virol 89:428–442CrossRefGoogle Scholar
- Nicoll MP, Hann W, Shivkumar M, Harman LE, Connor V, Coleman HM, Proenca JT, Efstathiou S (2016) The HSV-1 latency-associated transcript functions to repress latent phase lytic gene expression and suppress virus reactivation from latently infected neurons. PLoS Pathog 12:e1005539CrossRefGoogle Scholar
- Powell S, Vinod A, Lemons ML (2014) Isolation and culture of dissociated sensory neurons from chick embryos J Vis Exp 51991Google Scholar