Cellular gene expression survey of PseudoRabies Virus (PRV) infected Human Embryonic Kidney cells (HEK-293)

Abstract

-Pseudorabies virus (PRV)is an alpha herpesvirus that causes Aujezsky disease in the pig. To characterize the impact of PRV infection on cellular expression, we used microarrays consisting of 9850 oligonucleotides corresponding to human genes and examined the expression levels of mRNA isolated 0.5, 3, 6, and 9 h post infection (hpi) from cultures of infected HEK-293 cells. Very few changes were observed during the first 3 h of infection but significant modifications in the cell expression of more than 1000 genes were clearly apparent by 6 hpi. More than 2400 genes were either up-or down-regulated during the 9 h experiment. These results were then analyzed using gene ontology and the MAPP and MAPPFinder software. This comprehensive analysis clearly shows that the down-regulated genes were mainly involved in macromolecular synthesis (DNA, RNA and proteins) and the cell cycle. The up-regulated genes primarily concerned the regulation of DNA transcription, developmental processes (central nervous system development, neurogenesis, angiogenesis), cell adhesion and potassium transport. This study is the first qualitative analysis of a gene expression survey in a human cell line following PRV infection. It demonstrates global changes in the cell expression profile, and identifies the main biological processes that are altered during virus replication. pseudorabies virus / microarray / host pathogen interaction

A trypsin-like serine protease is involved in pseudorabies virus invasion through the basement membrane barrier of porcine nasal respiratory mucosa

“…An important question that remains is whether the proteases involved are of cellular and/or viral origin. Interestingly, transcriptome studies have indicated that several cellular components involved in trypsin-like serine protease activity are differentially regulated upon infection with alphaherpesviruses like PRV and HSV, including factor IX, tissue-type plasminogen activator (t-PA) and urinary plasminogen activator receptor 1 (uPAR1) [30,31]. Further research will be aimed at identifying the proteases involved and elucidating the exact molecular mechanism of viral penetration through the BM.…”

Transcriptomic analysis of the dialogue between Pseudorabies virus and porcine epithelial cells during infection

“…During recent years, DNA microarray technology has proven to be a very efficient high-throughput tool to study the gene expression profiles of infected host cells or pathogens [ 10 , 11 ]. To date, three transcriptomic analyses focused on cellular gene expression have been carried out in non-porcine PrV infected cells [ 12 - 14 ]. Ray and Enquist have compared the cellular pathways regulated by PrV and HSV-1 during infection of rat embryonic fibroblast cells using a rat microarray [ 12 ].…”

Quantitative Whole-Cell Proteome Analysis of Pseudorabies Virus-Infected Cells

“…In contrast to HSV-1 ICP27, the PrV homolog pUL54 is dispensable for virus replication in cell culture (47,49). Transcript analyses of PrV-infected rat (44), human (6), and porcine (12) cells demonstrated alterations in the abundance of individual cellular transcripts, which resulted in the depletion or accumulation of specific mRNAs. Of the 9,850 genes examined in one study (6), the number of significantly up-or downregulated genes increased from approximately 1,000 to over 2,400 between 6 and 9 h after infection.…”

“…Transcript analyses of PrV-infected rat (44), human (6), and porcine (12) cells demonstrated alterations in the abundance of individual cellular transcripts, which resulted in the depletion or accumulation of specific mRNAs. Of the 9,850 genes examined in one study (6), the number of significantly up-or downregulated genes increased from approximately 1,000 to over 2,400 between 6 and 9 h after infection. Evaluation of the functions annotated for highly regulated cellular genes shows that PrV infection influences numerous cellular pathways and that genes involved in protein and nucleic acid metabolism, signaling, transport, cell cycle control, adhesion, transcription, the stress response, and innate immunity are affected most frequently.…”