内含子中大潜伏转录物全长 microRNA 簇与伪狂犬病病毒的毒力相关。

The full-length microRNA cluster in the intron of large latency transcript is associated with the virulence of pseudorabies virus.

机构信息

State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.

出版信息

Virology. 2018 Jul;520:59-66. doi: 10.1016/j.virol.2018.05.004. Epub 2018 May 16.

DOI:10.1016/j.virol.2018.05.004

PMID:29777914

Abstract

Pseudorabies virus (PRV), the etiological pathogen of Aujeszky's disease, belongs to the Alphaherpesvirus subfamily. Large latency transcript (LLT), the most abundant PRV transcript, harbors a ~ 4.6 kb microRNA (miRNA) cluster-encoding intron. To investigate the function of the LLT miRNA cluster during the life cycle of PRV, we generated a miRNA cluster mutation virus (PRV-∆miR cluster) and revertant virus. Analysis of the growth kinetics of PRV-ΔmiR cluster-infected cells revealed significantly smaller plaques and lower titers than the wild-type and revertant viruses. The mutation virus exhibited increased IE180 and decreased EP0 expression. The clinical symptoms observed in mice infected with PRV-ΔmiR cluster revealed that the miRNA cluster is involved in the pathogenesis of PRV. Physical parameters, virus shedding assays, and the SN titers revealed that the miRNA cluster enhances PRV virulence in pigs. Collectively, our findings suggest that the full-length miRNA cluster is involved in PRV replication and virulence.

摘要

伪狂犬病病毒（PRV）是 Aujeszky 病的病原体，属于α疱疹病毒亚科。潜伏转录本（LLT）是 PRV 最丰富的转录本，含有一个约 4.6kb 的 microRNA（miRNA）簇编码内含子。为了研究 LLT miRNA 簇在 PRV 生命周期中的功能，我们生成了 miRNA 簇突变病毒（PRV-∆miR cluster）和回复病毒。分析 PRV-ΔmiR cluster 感染细胞的生长动力学，发现与野生型和回复病毒相比，其噬菌斑更小，滴度更低。突变病毒表现出 IE180 增加和 EP0 表达减少。感染 PRV-ΔmiR cluster 的小鼠的临床症状表明，miRNA 簇参与了 PRV 的发病机制。通过对物理参数、病毒脱落试验和 SN 滴度的检测，发现 miRNA 簇增强了 PRV 在猪体内的毒力。综上所述，我们的研究结果表明全长 miRNA 簇参与了 PRV 的复制和毒力。