Project description:RNA isolated from draining tracheobronchial lymph nodes (TBLN) from 5-week old pigs, either clinically infected with a feral isolate of Pseudorabies virus or uninfected were interrogated using Illumina Digital Gene Expression Tag Profiling. Over 100 million tag sequences were observed, representing 4,064,189 unique 21-base sequences collected from TBLN at time points 1, 3, 6 and 14 days post-infection (dpi)
Project description:RNA isolated from draining tracheobronchial lymph nodes (TBLN) from 5-week old pigs, either clinically infected with a feral isolate of Pseudorabies virus or uninfected were interrogated using Illumina Digital Gene Expression Tag Profiling. Over 100 million tag sequences were observed, representing 4,064,189 unique 21-base sequences collected from TBLN at time points 1, 3, 6 and 14 days post-infection (dpi) RNA isolated from draining tracheobronchial lymph nodes (TBLN) from 5-week old pigs (% per group pooled), either clinically infected with feral isolate FS268 of Pseudorabies virus or uninfected at 1, 3, 6, and 14 days post inoculation. Over 100 million tag sequences were observed, representing 4,064,189 unique 21-base sequences.
Project description:Six piglets (age 2-4 days) suffering from pseudorabies virus were identified during at outbreak at a commercial farm. RNA was extracted from lung (n=4) and brain (n=6) of the infected pigs, and compared to lung/brain RNA from a control pool of three uninfected piglets. Expression profiling used a human oligonucleotide geneset.
Project description:Pseudorabies, an acute infectious disease caused by pseudorabies virus (PRV), has caused enormous economic losses to the breeding industry in many countries worldwide. Accumulating evidence indicates that long non-coding RNAs (lncRNAs) may play important roles in the antiviral responses. However, little is known about the identification and functions of swine lncRNAs in cellular antiviral responses against PRV II. In this study, we detected the expression profiles of host lncRNAs and mRNAs from PRV-DX, a wild-type (WT) strain of PRV II, and its attenuated gE-TK- PRV-DX infected cells by high-throughput RNA sequencing. Finally, we identified differentially expressed (DE) 664 lncRNAs and DE 7,199 mRNAs from PRV-DX infected cells, and 654 DE lncRNAs and DE 7,149 mRNAs from gE-TK- PRV-DX infected cells when compared to MOCK infected cells, respectively, especially including 469 common DE lncRNAs and 5836 DE mRNAs. Besides 276 DE lncRNAs and 2,272 DE mRNAs between PRV-DX and gE-TK- PRV-DX infected cells were identified. The potential functions of the significant DE lncRNAs were involved in interleukin secretion, axon extension and metabolic process based on the Gene ontology and Kyoto Encyclopedia of Genes and Genomes databases. Taken together, results highlighted the potentials of lncRNA as targets for antiviral therapy and enriched the current knowledge of the mechanisms underlying the interaction between PRV II and its host cells.
Project description:Pseudorabies virus (PRV), the etiological agent of Aujeszky’s disease, is a swine pathogen of the alphaherpesvirinae subfamily resulting in devastating neurological and respiratory disorders in piglets, or abortion in pregnant sows.Here, we performed isobaric tags for relative and absolute quantitation (iTRAQ) quantitative phosphoproteomics on PRV-infected PK-15 cells, which identified 5723 phospho-peptides, corresponding to 2180 proteins, including 810 proteins showing significant changes in phosphorylation level during early PRV infection.
Project description:BackgroundPseudorabies virus is a widely-studied model organism of the Herpesviridae family, with a compact genome arrangement of 72 known coding sequences. In order to obtain an up-to-date genetic map of the virus, a combination of RNA-sequencing approaches were applied, as recent advancements in high-throughput sequencing methods have provided a wealth of information on novel RNA species and transcript isoforms, revealing additional layers of transcriptome complexity in several viral species.ResultsThe total RNA content and polyadenylation landscape of pseudorabies virus were characterized for the first time at high coverage by Illumina high-throughput sequencing of cDNA samples collected during the lytic infectious cycle. As anticipated, nearly all of the viral genome was transcribed, with the exception of loci in the large internal and terminal repeats, and several small intergenic repetitive sequences. Our findings included a small novel polyadenylated non-coding RNA near an origin of replication, and the single-base resolution mapping of 3' UTRs across the viral genome. Alternative polyadenylation sites were found in a number of genes and a novel alternative splice site was characterized in the ep0 gene, while previously known splicing events were confirmed, yielding no alternative splice isoforms. Additionally, we detected the active polyadenylation of transcripts earlier believed to be transcribed as part of polycistronic RNAs.ConclusionTo the best of our knowledge, the present work has furnished the highest-resolution transcriptome map of an alphaherpesvirus to date, and reveals further complexities of viral gene expression, with the identification of novel transcript boundaries, alternative splicing of the key transactivator EP0, and a highly abundant, novel non-coding RNA near the lytic replication origin. These advances provide a detailed genetic map of PRV for future research.
Project description:In the last couple of years, the implementation of long-read sequencing (LRS) technologies for transcriptome profiling has uncovered an extreme complexity of viral gene expression. In this study, we carried out a systematic analysis on the pseudorabies virus transcriptome by combining our current data obtained by using Pacific Biosciences Sequel and Oxford Nanopore Technologies MinION sequencing with our earlier data generated by other LRS and short-read sequencing techniques. As a result, we identified a number of novel genes, transcripts, and transcript isoforms, including splice and length variants, and also confirmed earlier annotated RNA molecules. One of the major findings of this study is the discovery of a large number of 5'-truncations of larger putative mRNAs being 3'-co-terminal with canonical mRNAs of PRV. A large fraction of these putative RNAs contain in-frame ATGs, which might initiate translation of N-terminally truncated polypeptides. Our analyses indicate that CTO-S, a replication origin-associated RNA molecule is expressed at an extremely high level. This study demonstrates that the PRV transcriptome is much more complex than previously appreciated.
Project description:Alpha-herpesviruses establish a life-long infection in the nervous system of the affected host; while this infection is restricted to peripheral neurons in a heathy host, it can spread within the neuronal circuitry in compromised individuals leading to adverse health consequences. Pseudorabies virus (PRV) an alpha-herpesvirus, requires the viral protein Us9 for sorting virus particles into axons. It does so by mediating an interaction of virus particles with neuronal transport machinery. Us9-mediated axonal sorting also depends on the state of neuronal maturation as the proteome composition changes with neuronal development of dendrites and axons. Immature superior cervical ganglia (SCGs) have rudimentary neurites that lack markers of mature dendrites or axons. Immature SCGs can be infected by PRV, but show markedly reduced Us9-dependent sorting into neurites. Mature SCGs abundantly express a variety of proteins characteristic of vesicle-transport machinery. Proteomics studies identified several novel Us9-associated neuronal proteins with potential roles in the regulation of axonal sorting and subsequent anterograde spread of virus particles in axons. For example, we found that SMPD4/n-sMase3, a sphingomyelinase abundant in lipid-rafts, associates with Us9 and is a negative regulator of axonal mediated spread of PRV, a potential novel antiviral function.