The architecture of the simian varicella virus transcriptome.
The architecture of the simian varicella virus transcriptome.
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Simian Vericella病毒转录组的结构。
DOI：
10.1371/journal.ppat.1010084
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发表时间：
2021-11
期刊：
影响因子：
6.7
通讯作者：
Ouwendijk WJD
中科院分区：
文献类型：
--
作者：
Braspenning SE;Verjans GMGM;Mehraban T;Messaoudi I;Depledge DP;Ouwendijk WJD
水痘带状疱疹病毒（VZV）的原发性感染导致水痘和感觉神经节神经元中终身潜伏期的建立。在三分之一的感染者中，VZV从潜伏期重新激活，引起带状疱疹，通常伴有难以治疗的慢性疼痛。猴水痘病毒（SVV）对非人灵长类动物的实验感染再现了人类VZV病的大部分特征，从而为研究水痘和带状疱疹的体内发病机制提供了机会。然而，与VZV相比，SVV的转录组和完整的编码潜力仍然不完全清楚。在这里，我们进行了纳米孔直接RNA测序，以注释SVV裂解感染的非洲绿色猴（AGM）和恒河猴（RM）肾上皮细胞中的SVV转录组。我们完善了经典SVV转录本的结构，并发现了许多RNA亚型，剪接事件，融合转录本和非编码RNA，大多数是SVV所特有的。我们验证了典型的和新发现的SVV转录在体内的表达，使用急性SVV感染的食蟹猴的肺样本。通过逆转录PCR确认所选转录物同种型的表达，包括位于SVV基因组独特左端的那些。最后，我们进行了详细的表征SVV同源的VZV潜伏相关的转录本（VLT），位于ORF 61的反义。与VZV VLT类似，SVV VLT是多重剪接的，并且使用替代转录起始位点和广泛剪接产生许多同种型。相反，单个剪接的SVV VLT同种型的低水平表达定义了体内潜伏期。值得注意的是，VLT核心外显子的基因组位置在SVV和VZV之间高度保守。因此，这项工作突出了裂解SVV基因表达的复杂性，并提供了新的见解裂解和潜伏SVV感染的分子生物学基础。SVV VLT同源物的鉴定进一步强调了SVV非人灵长类动物模型在开发预防带状疱疹的新策略中的价值。水痘-带状疱疹病毒（VZV）-一种普遍存在的人类病原体-在儿童时期感染大多数个体，导致水痘，之后病毒在宿主中持续数十年。在以后的生活中，VZV重新激活引起带状疱疹，经常与难以治疗的慢性疼痛有关。我们对病毒生命周期的有限了解阻碍了更有效治疗方案的开发。猿猴水痘病毒（SVV）是VZV的非人灵长类同源物，并且在旧世界猴中引起具有类似于人VZV感染的临床、病理和免疫学特征的自然疾病。然而，目前还不清楚这两种病毒在分子水平上有多相似。在这里，我们已经重新审视了SVV的全基因组的转录活性溶菌感染的肾上皮细胞来自两个非人灵长类动物物种和验证表达的新发现的病毒转录本在肺组织中SVV感染的动物。总之，这导致了许多替代RNA亚型的鉴定，其中大部分是SVV特有的，其中一些可能对病毒具有功能性影响。值得注意的是，我们定义了SVV潜伏相关转录本，它与VZV对应物高度相似。总之，我们的研究显示了理解给定动物模型的分子生物学的价值，并确定了潜在的保守的潜伏机制。
Primary infection with varicella-zoster virus (VZV) causes varicella and the establishment of lifelong latency in sensory ganglion neurons. In one-third of infected individuals VZV reactivates from latency to cause herpes zoster, often complicated by difficult-to-treat chronic pain. Experimental infection of non-human primates with simian varicella virus (SVV) recapitulates most features of human VZV disease, thereby providing the opportunity to study the pathogenesis of varicella and herpes zoster in vivo. However, compared to VZV, the transcriptome and the full coding potential of SVV remains incompletely understood. Here, we performed nanopore direct RNA sequencing to annotate the SVV transcriptome in lytically SVV-infected African green monkey (AGM) and rhesus macaque (RM) kidney epithelial cells. We refined structures of canonical SVV transcripts and uncovered numerous RNA isoforms, splicing events, fusion transcripts and non-coding RNAs, mostly unique to SVV. We verified the expression of canonical and newly identified SVV transcripts in vivo, using lung samples from acutely SVV-infected cynomolgus macaques. Expression of selected transcript isoforms, including those located in the unique left-end of the SVV genome, was confirmed by reverse transcription PCR. Finally, we performed detailed characterization of the SVV homologue of the VZV latency-associated transcript (VLT), located antisense to ORF61. Analogous to VZV VLT, SVV VLT is multiply spliced and numerous isoforms are generated using alternative transcription start sites and extensive splicing. Conversely, low level expression of a single spliced SVV VLT isoform defines in vivo latency. Notably, the genomic location of VLT core exons is highly conserved between SVV and VZV. This work thus highlights the complexity of lytic SVV gene expression and provides new insights into the molecular biology underlying lytic and latent SVV infection. The identification of the SVV VLT homolog further underlines the value of the SVV non-human primate model to develop new strategies for prevention of herpes zoster. Varicella-zoster virus (VZV)–a ubiquitous human pathogen–infects most individuals during childhood, leading to chickenpox, after which the virus persists in the host for decades. Later in life, VZV reactivates to cause shingles, frequently associated with difficult-to-treat chronic pain. Our limited understanding of the viral life-cycle hampers the development of more effective treatment options. Simian varicella virus (SVV) is the non-human primate homologue of VZV and causes a natural disease in Old World monkeys with clinical, pathological, and immunological features resembling human VZV infection. However, it is unclear how similar both viruses are at the molecular level. Here, we have revisited the genome-wide transcriptional activity of SVV during lytic infection of kidney epithelial cells derived from two non-human primate species and validated expression of newly identified viral transcripts in lung tissue from SVV-infected animals. Together, this has led to the identification of numerous alternative RNA isoforms, mostly unique to SVV, and some of which may have functional implications for the virus. Notably, we defined the SVV latency-associated transcript, which is highly similar to its VZV counterpart. In conclusion, our study shows the value of understanding the molecular biology of a given animal model and identifies potentially conserved mechanism of latency.
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影响因子：
3.2
作者：
Ouwendijk, Werner J. D.;Mahalingam, Ravi;Traina-Dorge, Vicki;van Amerongen, Geert;Wellish, Mary;Osterhaus, Albert D. M. E.;Gilden, Don;Verjans, Georges M. G. M.
通讯作者：
Verjans, Georges M. G. M.
水痘带状疱疹病毒潜伏期的分子方面。
DOI：
10.3390/v10070349
发表时间：
2018-06-28
期刊：
Viruses
影响因子：
--
作者：
Depledge DP;Sadaoka T;Ouwendijk WJD
通讯作者：
Ouwendijk WJD
影响因子：
5.4
作者：
Che, Xibing;Reichelt, Mike;Arvin, Ann M.
通讯作者：
Arvin, Ann M.
影响因子：
7
作者：
Braunschweig U;Barbosa-Morais NL;Pan Q;Nachman EN;Alipanahi B;Gonatopoulos-Pournatzis T;Frey B;Irimia M;Blencowe BJ
通讯作者：
Blencowe BJ
影响因子：
5.4
作者：
Arnold, Nicole;Girke, Thomas;Messaoudia, Ilhem
通讯作者：
Messaoudia, Ilhem