Mamestra configurata nucleopolyhedrovirus A
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PROVIDER: PRJNA309548 | ENA |
REPOSITORIES: ENA
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Project description:Infection of lepidopterans by baculoviruses has been traditionally studied using in vitro systems which enable efficient and highly synchronous infection. Many studies using varied virus-host combinations have yielded great insight into the molecular processes by which these large double-stranded DNA viruses achieve infection of host cells. However, a key difference in the virus strategy for infection between individual hosts, and within an individual host, lies with the production of two different forms of the virus; occlusion derived virus, which enables primary infection of insect gut tissues and budded virus, which efficiently infects a variety of different insect tissues throughout the host. To examine the primary infection of midgut cells specifically, we used MacoNPV infection of Mamestra configurata fourth instar larvae as our model and measured the expression of viral genes over a time course of infection. Both digital PCR and RNA sequencing methods showed the profile of transcription to be different from those typically seen with in vitro methods, having unique collections of genes expressed early, as well as much greater expression of p6.9 and much reduced expression of polh and p10 late, in comparison with in vitro studies. These differences likely reflect unique characteristics of midgut cell infection, and provide clues as to the processes these viruses use to regulate expression of different viral forms used to access different host tissues. Baculoviruses are versatile DNA viruses with great potential both as gene therapy vehicles and as biological control tools. Extensive study of their transcriptome in vitro has yielded valuable tools for use in protein expression systems, however it is critical that we obtain a fuller understanding of their in vivo activities before their full medical and agricultural potential can be realized. In these studies we have assessed the gene expression program from a group II alphabaculovirus in the midgut of its complementary larval host and confirmed that the in vivo activities of baculoviruses are unique from what is known of their in vitro transcriptome. These studies provide a first foray with next-generation molecular tools into the characterization of baculovirus biology in vivo.
2016-01-23 | GSE77147 | GEO
Project description:Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) is the best-studied baculovirus and most commonly used virus vector for baculovirus expression vector systems. The effect of AcMNPV infection on host cells is incompletely understood. A microarray based on Spodoptera frugiperda ESTs was used to investigate the impact of AcMNPV on host gene expression in cultured S. frugiperda, Sf21 cells. Most host genes were down-regulated over the time course of infection, although a small number were up-regulated. The most highly up-regulated genes encoded heat shock protein 70s and several poorly characterized proteins. Regulated genes with the highest score identified by functional annotation clustering included primarily products required for protein expression and trafficking in the ER and golgi. All were significantly down-regulated by approximately 12h post-infection. Microarray data were validated by qRT-PCR. This study provides the first comprehensive host transcriptome overview of Sf21 cells during AcMNPV infection.
2014-08-26 | GSE60739 | GEO
Project description:To reveal dynamic regulation of host gene expression and an overview of events in the infection cycle after SpltNPV infection To analyze cellular responses to SpltNPV infection, a S.litura cell line (SL221) was infected with wild-type baculovirus SpltNPV at a MOI of 1. RNAs were isolated at three time points through the infection cycle (6hpi, 12hpi and 18hpi) and also from control (mock infected) cells. The obtained cellular ranscriptome from the host S.litura was set for illumina strand specific RNA sequencing (RNA-seq) to analyze changes in host cell gene expression upon SpltNPV infection.
2015-04-22 | E-GEOD-68037 | biostudies-arrayexpress
Project description:Mamestra configurata whole head transcriptome
| PRJNA596858 | ENA
Project description:Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) is the best-studied baculovirus and most commonly used virus vector for baculovirus expression vector systems. The effect of AcMNPV infection on host cells is incompletely understood. A microarray based on Spodoptera frugiperda ESTs was used to investigate the impact of AcMNPV on host gene expression in cultured S. frugiperda, Sf21 cells. Most host genes were down-regulated over the time course of infection, although a small number were up-regulated. The most highly up-regulated genes encoded heat shock protein 70s and several poorly characterized proteins. Regulated genes with the highest score identified by functional annotation clustering included primarily products required for protein expression and trafficking in the ER and golgi. All were significantly down-regulated by approximately 12h post-infection. Microarray data were validated by qRT-PCR. This study provides the first comprehensive host transcriptome overview of Sf21 cells during AcMNPV infection. Host gene expression of the Sf21 cells was measured in AcMNPV mock-infected Sf21cells as well as AcMNPV-infected Sf21 cells at 6, 12, and 24 hours post infection (hpi). Four independent experiments were performed at each time (6, 12, and 24 hpi) using different donors for each experiment. Only two 24 hour sample data sets were used in the final analysis as two did not meet QC criteria.
2014-08-26 | E-GEOD-60739 | biostudies-arrayexpress
Project description:Over the past decades, Helicoverpa armigera nucleopolyhedrovirus (HearNPV) has been widely used for biocontrol of cotton bollworm, which is one of the most destructive pest insects in agriculture worldwide. However, the molecular mechanism underlying the interaction between HearNPV and host insects remains poorly understood. In this study, high throughput RNA-sequencing was integrated with label-free quantitative proteomics analysis to examine the dynamics of gene expression in the fat body of H. armigera larvae in response to challenge with HearNPV. RNA-sequencing-based transcriptomic analysis indicated that host gene expression was substantially altered, yielding 3,850 differentially expressed genes (DEGs), while no global transcriptional shut-off effects were observed in the fat body. Among the DEGs, 60 immunity-related genes were down-regulated after baculovirus infection, a finding that was consistent with the results of quantitative real-time RT-PCR (qRT-PCR). Gene ontology and functional classification demonstrated that the majority of down-regulated genes were enriched in gene cohorts involved in energy, carbohydrate, and amino acid metabolic pathways. Proteomics analysis identified differentially expressed proteins in the fat body, among which 76 were up-regulated, whereas 373 were significantly down-regulated upon infection. The down-regulated proteins are involved in metabolic pathways such as energy metabolism, carbohydrate metabolism (CM), and amino acid metabolism, in agreement with the RNA-seq data. Furthermore, correlation analysis suggested a strong association between the mRNA level and protein abundance in the H. armigera fat body. More importantly, the predicted gene interaction network indicated that a large subset of metabolic networks was significantly negatively regulated by viral infection, including CM-related enzymes such as aldolase, enolase, malate dehydrogenase and triose-phosphate isomerase. Taken together, transcriptomic combined with proteomic data elucidated that baculovirus established systemic infection of host larvae, and manipulated the host mainly by suppressing the host immune response and down-regulating metabolism to allow viral self-replication and proliferation. Therefore, this study provided important insights into the mechanism of host-baculovirus interaction.
2017-04-19 | PXD004748 | Pride
Project description: Not available
| S-EPMC4825930 | biostudies-literature
Project description:Baculovirus expression systems have been widely used to produce recombinant mammalian proteins owing to the lack of viral replication in vertebrates. Although several lines of evidence have demonstrated impacts of baculovirus infection in mammalian hosts, genome-wide effects have not been fully elucidated. Here, we provide comparative transcriptome profiles of baculovirus and host-immune response genes in recombinant baculovirus-infected mammalian and insect cells. Specifically, to decipher the impacts of baculovirus infection in mammalian cells, we conducted total RNA-seq on human 293 cells and insect Sf9 cells infected with recombinant baculovirus. We found that baculovirus genes were rarely expressed under the control of baculoviral promoters in 293 cells. Although some baculovirus early genes, such as PE38 and IE-01, showed limited expression in 293 cells, baculoviral late genes were mostly silent. We also found modest induction of a small number of mammalian immune response genes associated with Toll-like receptors, cytokine signaling, and complement in baculovirus-infected 293 cells. These comprehensive transcriptome data will contribute to improving recombinant baculovirus as tools for gene delivery, gene therapy, and vaccine development.
2020-09-28 | GSE158603 | GEO