Transcriptome Responses of the Host Trichoplusia ni to Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus
Transcriptome Responses of the Host Trichoplusia ni to Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus
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DOI：
10.1128/jvi.02243-14
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发表时间：
2014-12-01
影响因子：
5.4
通讯作者：
Blissard, Gary W.
中科院分区：
文献类型：
--
作者：
Chen, Yun-Ru;Zhong, Silin;Blissard, Gary W.
苜蓿银纹夜蛾核型多角体病毒（Autographa californica multiple nucleopolyhedrovirus，AcMNPV）对粉纹夜蛾细胞的感染导致了156个病毒基因的表达，并导致了细胞的急剧重塑。由于缺乏T.ni的参考基因组和转录组，细胞转录组如何响应病毒感染尚不清楚。我们使用了一个类似于60 Gb RNA测序（RNA-seq）的数据集，从感染和未感染的T。NI细胞，以产生并注释约70，322T.niunigenes（组装的转录物）的从头转录组组装，代表48小时感染周期。利用差异基因表达分析，我们发现大多数宿主转录本在感染后6 h（p.i.）以及感染的整个过程而T. NI单基因在早期（感染后0至6小时）上调，随后在感染周期的剩余时间内减少。此外，与代谢和应激反应相关的一小部分基因在感染后18和24 h显示转录水平显著升高。但此后下降。我们还研究了属于一些特定途径的基因的反应，包括应激反应，细胞凋亡，免疫和蛋白质运输。我们确定了在感染早期上调的特定途径成员。与AcMNPV表达的平行分析相结合，这些结果提供了一个广泛的和详细的视图如何杆状病毒感染影响宿主细胞转录组逃避细胞防御反应，修改细胞生物合成途径，并重塑细胞结构。重要的是，杆状病毒是昆虫特异性的DNA病毒，是高致病性的昆虫宿主。除了用于某些昆虫的生物防治之外，杆状病毒还用作许多生物技术应用的病毒载体，例如用于疫苗生产的哺乳动物细胞转导和蛋白质表达。虽然有相当多的信息感染细胞中的病毒基因表达，很少有人知道宿主细胞的杆状病毒感染的反应。在这些研究中，我们从宿主粉纹夜蛾中组装了一个细胞转录组，并使用该转录组分析了整个感染周期中宿主细胞基因表达的变化。该研究与先前的病毒基因表达变化研究平行进行。结合起来，这些研究提供了前所未有的新水平的细节和感染周期中事件的概述，它们将刺激新的实验方法来理解，修改和利用杆状病毒的各种应用。
Productive infection of Trichoplusia ni cells by the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) leads to expression of similar to 156 viral genes and results in dramatic cell remodeling. How the cell transcriptome responds to viral infection was unknown due to the lack of a reference genome and transcriptome for T.ni. We used an similar to 60-Gb RNA sequencing (RNA-seq) data set from infected and uninfected T. ni cells to generate and annotate a de novo transcriptome assembly of approximately 70,322 T.ni unigenes (assembled transcripts), representing the 48-h infection cycle. Using differential gene expression analysis, we found that the majority of host transcripts were downregulated after 6 h postinfection (p.i.) and throughout the remainder of the infection. In contrast, 5.7% (4,028) of the T. ni unigenes were upregulated during the early period (0 to 6 h p.i.), followed by a decrease through the remainder of the infection cycle. Also, a small subset of genes related to metabolism and stress response showed a significant elevation of transcript levels at 18 and 24 h p.i. but a decrease thereafter. We also examined the responses of genes belonging to a number of specific pathways of interest, including stress responses, apoptosis, immunity, and protein trafficking. We identified specific pathway members that were upregulated during the early phase of the infection. Combined with the parallel analysis of AcMNPV expression, these results provide both a broad and a detailed view of how baculovirus infection impacts the host cell transcriptome to evade cellular defensive responses, to modify cellular biosynthetic pathways, and to remodel cell structure.IMPORTANCEBaculoviruses are insect-specific DNA viruses that are highly pathogenic to their insect hosts. In addition to their use for biological control of certain insects, baculoviruses also serve as viral vectors for numerous biotechnological applications, such as mammalian cell transduction and protein expression for vaccine production. While there is considerable information regarding viral gene expression in infected cells, little is known regarding responses of the host cell to baculovirus infection. In these studies, we assembled a cell transcriptome from the host Trichoplusia ni and used that transcriptome to analyze changes in host cell gene expression throughout the infection cycle. The study was performed in parallel with a prior study of changes in viral gene expression. Combined, these studies provide an unprecedented new level of detail and an overview of events in the infection cycle, and they will stimulate new experimental approaches to understand, modify, and utilize baculoviruses for a variety of applications.