Markus Amos, Golani Linoy, Ojha Nishant Kumar, Borodiansky-Shteinberg Tatiana, Kinchington Paul R, Goldstein Ronald S
Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
Departments of Ophthalmology and Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
J Virol. 2017 Nov 30;91(24). doi: 10.1128/JVI.01710-17. Print 2017 Dec 15.
Many herpesviruses express small noncoding RNAs (sncRNAs), including microRNAs (miRNAs), that may play roles in regulating lytic and latent infections. None have yet been reported in varicella-zoster virus (VZV; also known as human herpesvirus 3 [HHV-3]). Here we analyzed next-generation sequencing (NGS) data for small RNAs in VZV-infected fibroblasts and human embryonic stem cell-derived (hESC) neurons. Two independent bioinformatics analyses identified more than 20 VZV-encoded 20- to 24-nucleotide RNAs, some of which are predicted to have stem-loop precursors potentially representing miRNAs. These sequences are perfectly conserved between viruses from three clades of VZV. One NGS-identified sequence common to both bioinformatics analyses mapped to the repeat regions of the VZV genome, upstream of the predicted promoter of the immediate early gene open reading frame 63 (ORF63). This miRNA candidate was detected in each of 3 independent biological repetitions of NGS of RNA from fibroblasts and neurons productively infected with VZV using TaqMan quantitative PCR (qPCR). Importantly, transfected synthetic RNA oligonucleotides antagonistic to the miRNA candidate significantly enhanced VZV plaque growth rates. The presence of 6 additional small noncoding RNAs was also verified by TaqMan qPCR in productively infected fibroblasts and ARPE19 cells. Our results show VZV, like other human herpesviruses, encodes several sncRNAs and miRNAs, and some may regulate infection of host cells. Varicella-zoster virus is an important human pathogen, with herpes zoster being a major health issue in the aging and immunocompromised populations. Small noncoding RNAs (sncRNAs) are recognized as important actors in modulating gene expression, and this study demonstrates the first reported VZV-encoded sncRNAs. Many are clustered to a small genomic region, as seen in other human herpesviruses. At least one VZV sncRNA was expressed in productive infection of neurons and fibroblasts that is likely to reduce viral replication. Since sncRNAs have been suggested to be potential targets for antiviral therapies, identification of these molecules in VZV may provide a new direction for development of treatments for painful herpes zoster.
许多疱疹病毒表达小非编码RNA（sncRNA），包括微小RNA（miRNA），它们可能在调节裂解性感染和潜伏性感染中发挥作用。水痘带状疱疹病毒（VZV；也称为人类疱疹病毒3型[HHV-3]）中尚未有相关报道。在此，我们分析了VZV感染的成纤维细胞和人胚胎干细胞来源（hESC）神经元中小RNA的下一代测序（NGS）数据。两项独立的生物信息学分析鉴定出20多种VZV编码的20至24个核苷酸的RNA，其中一些预计具有可能代表miRNA的茎环前体。这些序列在VZV三个进化枝的病毒之间完全保守。两项生物信息学分析共同鉴定出的一个NGS序列映射到VZV基因组的重复区域，位于立即早期基因开放阅读框63（ORF63）预测启动子的上游。使用TaqMan定量PCR（qPCR）在来自成纤维细胞和神经元的RNA的3次独立生物学重复的NGS中，每次都检测到了这种miRNA候选物，这些细胞被VZV有效感染。重要的是，转染与该miRNA候选物拮抗的合成RNA寡核苷酸可显著提高VZV斑块生长速率。另外6种小非编码RNA的存在也通过TaqMan qPCR在有效感染的成纤维细胞和ARPE19细胞中得到验证。我们的结果表明，VZV与其他人类疱疹病毒一样，编码多种sncRNA和miRNA，其中一些可能调节宿主细胞的感染。水痘带状疱疹病毒是一种重要的人类病原体，带状疱疹是老年和免疫功能低下人群中的一个主要健康问题。小非编码RNA（sncRNA）被认为是调节基因表达的重要因素，本研究首次报道了VZV编码的sncRNA。许多sncRNA聚集在一个小的基因组区域，这与其他人类疱疹病毒的情况相同。至少一种VZV sncRNA在神经元和成纤维细胞的有效感染中表达，并可能减少病毒复制。由于sncRNA已被认为是抗病毒治疗的潜在靶点，在VZV中鉴定出这些分子可能为开发治疗带状疱疹疼痛的新方法提供新方向。
