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Lynn W. Enquist - One of the best experts on this subject based on the ideXlab platform.
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the axonal sorting activity of Pseudorabies Virus us9 protein depends on the state of neuronal maturation
bioRxiv, 2020Co-Authors: Nikhila S Tanneti, Ileana M Cristea, Joel D Federspiel, Lynn W. EnquistAbstract:Abstract 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 healthy host, the reactivated Virus can spread within the neuronal circuitry, such as to the brain, in compromised individuals and lead to adverse health outcomes. Pseudorabies Virus (PRV), an alpha-herpesVirus, requires the viral protein Us9 to sort Virus particles into axons and facilitate neuronal spread. Us9 sorts Virus particles by mediating the interaction of Virus particles with neuronal transport machinery. Here, we report that Us9-mediated regulation of axonal sorting also depends on the state of neuronal maturation. Specifically, the development of dendrites and axons is accompanied with proteomic changes that influence neuronal processes. Immature superior cervical ganglionic neurons (SCGs) have rudimentary neurites that lack markers of mature axons. Immature SCGs can be infected by PRV, but they show markedly reduced Us9-dependent regulation of sorting, and increased Us9-independent transport of particles into neurites. Mature SCGs have relatively higher abundances of proteins characteristic of vesicle-transport machinery. We also identify Us9-associated neuronal proteins that can contribute to axonal sorting and subsequent anterograde spread of Virus particles in axons. We show that SMPD4/nsMase3, a sphingomyelinase abundant in lipid-rafts, associates with Us9 and is a negative regulator of PRV sorting into axons and neuronal spread, a potential antiviral function. Author Summary Viral pathogenesis often is age-dependent, with more severe outcomes for infected fetuses and neonates compared to adults. As neurons age and mature, dendrites and axons polarize with distinct functions that affect neurotropic Virus replication and neuronal spread of infection. This study investigates how neuronal maturation of peripheral nervous system neurons, the site of alpha-herpesVirus life-long latency and reactivation, affects replication and neuronal spread of Pseudorabies Virus. Characterization of infected immature and mature primary cultures of superior cervical ganglionic neurons revealed significant differences in protein composition and cellular processes that affected the activity of Us9, a viral protein required for sorting Virus particles into axons. We identified neuronal and viral proteins that interact with Us9 in immature and mature neurons. Among these, we demonstrate that SMPD4/nsMase3, a sphingomyelinase critical for membrane organization and neuronal function, regulates PRV neuronal spread by preventing capsid association with Us9-containing membranes, presenting a possible antiviral function.
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Pseudorabies Virus Infection Accelerates Degradation of the Kinesin-3 Motor KIF1A.
Journal of virology, 2020Co-Authors: Hao Huang, Orkide O Koyuncu, Lynn W. EnquistAbstract:AlphaherpesViruses, including Pseudorabies Virus (PRV), are neuroinvasive pathogens that establish lifelong latency in peripheral ganglia following the initial infection at mucosal surfaces. The establishment of latent infection and subsequent reactivations, during which newly assembled virions are sorted into and transported anterogradely inside axons to the initial mucosal site of infection, rely on axonal bidirectional transport mediated by microtubule-based motors. Previous studies using cultured peripheral nervous system (PNS) neurons have demonstrated that KIF1A, a kinesin-3 motor, mediates the efficient axonal sorting and transport of newly assembled PRV virions. Here we report that KIF1A, unlike other axonal kinesins, is an intrinsically unstable protein prone to proteasomal degradation. Interestingly, PRV infection of neuronal cells leads not only to a nonspecific depletion of KIF1A mRNA but also to an accelerated proteasomal degradation of KIF1A proteins, leading to a near depletion of KIF1A protein late in infection. Using a series of PRV mutants deficient in axonal sorting and anterograde spread, we identified the PRV US9/gE/gI protein complex as a viral factor facilitating the proteasomal degradation of KIF1A proteins. Moreover, by using compartmented neuronal cultures that fluidically and physically separate axons from cell bodies, we found that the proteasomal degradation of KIF1A occurs in axons during infection. We propose that the PRV anterograde sorting complex, gE/gI/US9, recruits KIF1A to viral transport vesicles for axonal sorting and transport and eventually accelerates the proteasomal degradation of KIF1A in axons.IMPORTANCE Pseudorabies Virus (PRV) is an alphaherpesVirus related to human pathogens herpes simplex Viruses 1 and 2 and varicella-zoster Virus. AlphaherpesViruses are neuroinvasive pathogens that establish lifelong latent infections in the host peripheral nervous system (PNS). Following reactivation from latency, infection spreads from the PNS back via axons to the peripheral mucosal tissues, a process mediated by kinesin motors. Here, we unveil and characterize the underlying mechanisms for a PRV-induced, accelerated degradation of KIF1A, a kinesin-3 motor promoting the sorting and transport of PRV virions in axons. We show that PRV infection disrupts the synthesis of KIF1A and simultaneously promotes the degradation of intrinsically unstable KIF1A proteins by proteasomes in axons. Our work implies that the timing of motor reduction after reactivation would be critical because progeny particles would have a limited time window for sorting into and transport in axons for further host-to-host spread.
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the neuropathic itch caused by Pseudorabies Virus
Pathogenetics, 2020Co-Authors: Kathlyn Laval, Lynn W. EnquistAbstract:Pseudorabies Virus (PRV) is an alphaherpesVirus related to varicella-zoster Virus (VZV) and herpes simplex Virus type 1 (HSV1). PRV is the causative agent of Aujeskzy's disease in swine. PRV infects mucosal epithelium and the peripheral nervous system (PNS) of its host where it can establish a quiescent, latent infection. While the natural host of PRV is the swine, a broad spectrum of mammals, including rodents, cats, dogs, and cattle can be infected. Since the nineteenth century, PRV infection is known to cause a severe acute neuropathy, the so called "mad itch" in non-natural hosts, but surprisingly not in swine. In the past, most scientific efforts have been directed to eradicating PRV from pig farms by the use of effective marker vaccines, but little attention has been given to the processes leading to the mad itch. The main objective of this review is to provide state-of-the-art information on the mechanisms governing PRV-induced neuropathic itch in non-natural hosts. We highlight similarities and key differences in the pathogenesis of PRV infections between non-natural hosts and pigs that might explain their distinctive clinical outcomes. Current knowledge on the neurobiology and possible explanations for the unstoppable itch experienced by PRV-infected animals is also reviewed. We summarize recent findings concerning PRV-induced neuroinflammatory responses in mice and address the relevance of this animal model to study other alphaherpesVirus-induced neuropathies, such as those observed for VZV infection.
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Pseudorabies Virus infection accelerates degradation of the kinesin 3 motor kif1a
bioRxiv, 2019Co-Authors: Hao Huang, Orkide O Koyuncu, Lynn W. EnquistAbstract:AlphaherpesViruses, including Pseudorabies Virus (PRV), are neuroinvasive pathogens that establish life-long latency in peripheral ganglia following the initial infection at mucosal surfaces. The establishment of latent infection and the subsequent reactivations during which newly-assembled virions are sorted into and transported anterogradely inside axons to the initial mucosal site of infection, rely on axonal bidirectional transport mediated by microtubule-based motors. Previous studies using cultured peripheral nervous system (PNS) neurons have demonstrated that KIF1A, a kinesin-3 motor, mediates the efficient axonal sorting and transport of newly-assembled PRV virions. In this study, we report that KIF1A, unlike other axonal kinesins, is an intrinsically unstable protein prone to proteasomal degradation. Interestingly, PRV infection of neuronal cells leads not only to a non-specific depletion of KIF1A mRNA, but also to an accelerated proteasomal degradation of KIF1A proteins, leading to a near depletion of KIF1A protein late in infection. Using a series of PRV mutants deficient in axonal sorting and anterograde spread, we identified the PRV US9/gE/gI protein complex as a viral factor facilitating the proteasomal degradation of KIF1A proteins. Moreover, by using compartmented neuronal cultures that fluidically and physically separate axons from cell bodies, we found that the proteasomal degradation of KIF1A occurs in axons during infection. We propose that PRV anterograde sorting complex, gE/gI/US9, recruits KIF1A to viral transport vesicles for axonal sorting and transport, and eventually accelerates the proteasomal degradation of KIF1A in axons.
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virulent Pseudorabies Virus infection induces a specific and lethal systemic inflammatory response in mice
Journal of Virology, 2018Co-Authors: Kathlyn Laval, Orkide O Koyuncu, J B Vernejoul, J Van Cleemput, Lynn W. EnquistAbstract:Pseudorabies Virus (PRV) is an alphaherpesVirus that infects the peripheral nervous system (PNS). The natural host of PRV is the swine, but it can infect most mammals, including cattle, rodents, and dogs. In these nonnatural hosts, PRV always causes a severe acute and lethal neuropathy called the “mad itch,” which is uncommon in swine. Thus far, the pathophysiological and immunological processes leading to the development of the neuropathic itch and the death of the animal are unclear. Using a footpad inoculation model, we established that mice inoculated with PRV-Becker (virulent strain) develop a severe pruritus in the foot and become moribund at 82 h postinoculation (hpi). We found necrosis and inflammation with a massive neutrophil infiltration only in the footpad and dorsal root ganglia (DRGs) by hematoxylin and eosin staining. PRV load was detected in the foot, PNS, and central nervous system tissues by quantitative reverse transcription-PCR. Infected mice had elevated plasma levels of proinflammatory cytokines (interleukin-6 [IL-6] and granulocyte colony-stimulating factor [G-CSF]) and chemokines (Gro-1 and monocyte chemoattractant protein 1). Significant IL-6 and G-CSF levels were detected in several tissues at 82 hpi. High plasma levels of C-reactive protein confirmed the acute inflammatory response to PRV-Becker infection. Moreover, mice inoculated with PRV-Bartha (attenuated, live vaccine strain) did not develop pruritus at 82 hpi. PRV-Bartha also replicated in the PNS, and the infection spread further in the brain than PRV-Becker. PRV-Bartha infection did not induce the specific and lethal systemic inflammatory response seen with PRV-Becker. Overall, we demonstrated the importance of inflammation in the clinical outcome of PRV infection in mice and provide new insights into the process of PRV-induced neuroinflammation. IMPORTANCE Pseudorabies Virus (PRV) is an alphaherpesVirus related to human pathogens such as herpes simplex Virus 1 and varicella-zoster Virus (VZV). The natural host of PRV is the swine, but it can infect most mammals. In susceptible animals other than pigs, PRV infection always causes a characteristic lethal pruritus known as the “mad itch.” The role of the immune response in the clinical outcome of PRV infection is still poorly understood. Here, we show that a systemic host inflammatory response is responsible for the severe pruritus and acute death of mice infected with virulent PRV-Becker but not mice infected with attenuated strain PRV-Bartha. In addition, we identified IL-6 and G-CSF as two main cytokines that play crucial roles in the regulation of this process. Our findings give new insights into neuroinflammatory diseases and strengthen further the similarities between VZV and PRV infections at the level of innate immunity.
Zsolt Boldogkői - One of the best experts on this subject based on the ideXlab platform.
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an integrated sequencing approach for updating the Pseudorabies Virus transcriptome
Pathogenetics, 2021Co-Authors: Gabor Torma, Dora Tombacz, Zsolt Csabai, Michael Snyder, Daniel Gobhardter, Zoltan Deim, Zsolt BoldogkőiAbstract: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.
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transcriptome wide survey of Pseudorabies Virus using next and third generation sequencing platforms
Scientific Data, 2018Co-Authors: Dora Tombacz, Donald Sharon, Michael Snyder, Attila Szűcs, Norbert Moldovan, Zsolt BoldogkőiAbstract:Pseudorabies Virus (PRV) is an alphaherpesVirus of swine. PRV has a large double-stranded DNA genome and, as the latest investigations have revealed, a very complex transcriptome. Here, we present a large RNA-Seq dataset, derived from both short- and long-read sequencing. The dataset contains 1.3 million 100 bp paired-end reads that were obtained from the Illumina random-primed libraries, as well as 10 million 50 bp single-end reads generated by the Illumina polyA-seq. The Pacific Biosciences RSII non-amplified method yielded 57,021 reads of inserts (ROIs) aligned to the viral genome, the amplified method resulted in 158,396 PRV-specific ROIs, while we obtained 12,555 ROIs using the Sequel platform. The Oxford Nanopore's MinION device generated 44,006 reads using their regular cDNA-sequencing method, whereas 29,832 and 120,394 reads were produced by using the direct RNA-sequencing and the Cap-selection protocols, respectively. The raw reads were aligned to the PRV reference genome (KJ717942.1). Our provided dataset can be used to compare different sequencing approaches, library preparation methods, as well as for validation and testing bioinformatic pipelines.
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multi platform sequencing approach reveals a novel transcriptome profile in Pseudorabies Virus
Frontiers in Microbiology, 2018Co-Authors: Norbert Moldovan, Dora Tombacz, Zsolt Csabai, Michael Snyder, Attila Szűcs, Zsolt BoldogkőiAbstract:Third-generation sequencing is an emerging technology that is capable of solving several problems that earlier approaches were not able to, including the identification of transcripts isoforms and overlapping transcripts. In this study, we used long-read sequencing for the analysis of Pseudorabies Virus (PRV) transcriptome, including Oxford Nanopore Technologies MinION, PacBio RS-II, and Illumina HiScanSQ platforms. We also used data from our previous short-read and long-read sequencing studies for the comparison of the results and in order to confirm the obtained data. Our investigations identified 19 formerly unknown putative protein-coding genes, all of which are 5’ truncated forms of earlier annotated longer PRV genes. Additionally, we detected 19 non-coding RNAs, including 5’ and 3’ truncated transcripts without in-frame ORFs, antisense RNAs, as well as RNA molecules encoded by those parts of the viral genome where no transcription had been detected before. This study has also led to the identification of three complex transcripts and fifty distinct length isoforms, including transcription start and end variants. We also detected 121 novel transcript overlaps, and two transcripts that overlap the replication origins of PRV. Furthermore, in silico analysis revealed 145 upstream ORFs, many of which are located on the longer 5’ isoforms of the transcripts.
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characterization of novel transcripts in Pseudorabies Virus
Viruses, 2015Co-Authors: Dora Tombacz, Donald Sharon, Peter Olah, Zsolt Csabai, Michael Snyder, Zoltan Havelda, Zsolt BoldogkőiAbstract:In this study we identified two 3'-coterminal RNA molecules in the Pseudorabies Virus. The highly abundant short transcript (CTO-S) proved to be encoded between the ul21 and ul22 genes in close vicinity of the replication origin (OriL) of the Virus. The less abundant long RNA molecule (CTO-L) is a transcriptional readthrough product of the ul21 gene and overlaps OriL. These polyadenylated RNAs were characterized by ascertaining their nucleotide sequences with the Illumina HiScanSQ and Pacific Biosciences Real-Time (PacBio RSII) sequencing platforms and by analyzing their transcription kinetics through use of multi-time-point Real-Time RT-PCR and the PacBio RSII system. It emerged that transcription of the CTOs is fully dependent on the viral transactivator protein IE180 and CTO-S is not a microRNA precursor. We propose an interaction between the transcription and replication machineries at this genomic location, which might play an important role in the regulation of DNA synthesis.
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strain kaplan of Pseudorabies Virus genome sequenced by pacbio single molecule real time sequencing technology
Genome Announcements, 2014Co-Authors: Dora Tombacz, Donald Sharon, Peter Olah, Zsolt Csabai, Michael Snyder, Zsolt BoldogkőiAbstract:Pseudorabies Virus (PRV) is a neurotropic herpesVirus that causes Aujeszky's disease in pigs. PRV strains are widely used as transsynaptic tracers for mapping neural circuits. We present here the complete and fully annotated genome sequence of strain Kaplan of PRV, determined by Pacific Biosciences RSII long-read sequencing technology.
Herman Favoreel - One of the best experts on this subject based on the ideXlab platform.
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Pseudorabies Virus inhibits type i and type iii interferon induced signaling via proteasomal degradation of janus kinases
Journal of Virology, 2021Co-Authors: Yue Yin, Nicolas Romero, Herman FavoreelAbstract:Both type I and III interferons (IFNs) play a crucial role in host antiviral response by activating the JAK/STAT (Janus kinase/signal transducer and activator of transcription) signaling pathway to trigger the expression of antiviral IFN-stimulated genes (ISGs). We report that the porcine alphaherpesVirus Pseudorabies Virus (PRV) triggers proteasomal degradation of the key Janus kinases Jak1 and to a lesser exent Tyk2, thereby inhibiting both type I and III IFN-induced STAT1 phosphorylation and suppressing IFN-induced expression of ISGs. UV-inactivated PRV did not interfere with IFN signaling. In addition, deletion of the EP0 gene from the PRV genome or inhibition of viral genome replication did not affect PRV-induced inhibition of IFN signaling. To our knowledge, this is the first report describing Janus kinase degradation by alphaherpesViruses. These findings thus reveal a novel alphaherpesVirus evasion mechanism of type I and type III IFNs. IMPORTANCE Type I and III IFNs trigger signaling via Janus kinases that phosphorylate and activate STAT transcription factors, leading to the expression of antiviral interferon-stimulated genes (ISGs) that result in an antiviral state of host cells. Viruses have evolved various mechanisms to evade this response. Our results indicate that an alphaherpesVirus, the porcine Pseudorabies Virus (PRV), inhibits both type I and III IFNs signaling pathways by triggering proteasome-dependent degradation of the key Janus kinases Jak1 and Tyk2 and consequent inhibition of STAT1 phosphorylation and suppression of ISG expression. Moreover, we found that this inhibition is not caused by incoming virions and does not depend on expression of the viral EP0 protein or viral true late proteins. These data for the first time address alphaherpesVirus evasion of type III IFN-mediated signaling and reveal a previously uncharacterized alphaherpesVirus mechanism of IFN evasion via proteasomal degradation of Janus kinases.
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Data_Sheet_2_Porcine NK Cells Stimulate Proliferation of Pseudorabies Virus-Experienced CD8+ and CD4+CD8+ T Cells.PDF
2019Co-Authors: Steffi De Pelsmaeker, Nick De Regge, Bert Devriendt, Herman FavoreelAbstract:Natural killer (NK) cells belong to the innate immune system and play a central role in the defense against viral infections and cancer development, but also contribute to shaping adaptive immune responses. NK cells are particularly important in the first line defense against herpesViruses, including alphaherpesViruses. In addition to their ability to kill target cells and produce interferon-γ, porcine and human NK cell subsets have been reported to display features associated with professional antigen presenting cells (APC), although it is currently unclear whether NK cells may internalize debris of Virus-infected cells and whether this APC-like activity of NK cells may stimulate proliferation of antiviral T cells. Here, using the porcine alphaherpesVirus Pseudorabies Virus (PRV), we show that vaccination of pigs with a live attenuated PRV vaccine strain triggers expression of MHC class II on porcine NK cells, that porcine NK cells can internalize debris from PRV-infected target cells, and that NK cells can stimulate proliferation of CD8+ and CD4+CD8+ PRV-experienced T cells. These results highlight the potential of targeting these NK cell features in future vaccination strategies.
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Porcine NK Cells Stimulate Proliferation of Pseudorabies Virus-Experienced CD8+ and CD4+CD8+ T Cells
Frontiers Media S.A., 2019Co-Authors: Steffi De Pelsmaeker, Nick De Regge, Bert Devriendt, Herman FavoreelAbstract:Natural killer (NK) cells belong to the innate immune system and play a central role in the defense against viral infections and cancer development, but also contribute to shaping adaptive immune responses. NK cells are particularly important in the first line defense against herpesViruses, including alphaherpesViruses. In addition to their ability to kill target cells and produce interferon-γ, porcine and human NK cell subsets have been reported to display features associated with professional antigen presenting cells (APC), although it is currently unclear whether NK cells may internalize debris of Virus-infected cells and whether this APC-like activity of NK cells may stimulate proliferation of antiviral T cells. Here, using the porcine alphaherpesVirus Pseudorabies Virus (PRV), we show that vaccination of pigs with a live attenuated PRV vaccine strain triggers expression of MHC class II on porcine NK cells, that porcine NK cells can internalize debris from PRV-infected target cells, and that NK cells can stimulate proliferation of CD8+ and CD4+CD8+ PRV-experienced T cells. These results highlight the potential of targeting these NK cell features in future vaccination strategies
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Pseudorabies Virus glycoprotein ge triggers erk1 2 phosphorylation and degradation of the pro apoptotic protein bim in epithelial cells
Virus Research, 2016Co-Authors: Maria Setas Lopes Pontes, Hans Nauwynck, Cliff Van Waesberghe, Bruno Verhasselt, Herman FavoreelAbstract:ERK1/2 (Extracellular signal Regulated Kinase 1/2) signaling is a key cellular signaling axis controlling many cellular events, including cell survival. Activation of ERK 1/2 may trigger an anti-apoptotic response, and different Viruses have been shown to benefit from this process. We have described recently that the viral glycoprotein gE mediates Pseudorabies Virus (PRV)-induced activation of ERK 1/2 in T lymphocytes. In the present study, we report that PRV gE-mediated ERK 1/2 phosphorylation also occurs in epithelial cells and that in these cells, gE-mediated ERK 1/2 signaling is associated with degradation of the pro-apoptotic protein Bim. Our results for the first time link the viral glycoprotein gE, an important alphaherpesVirus virulence factor, with the apoptotic signaling pathway.
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Pseudorabies Virus glycoprotein ge triggers erk1 2 phosphorylation and degradation of the pro apoptotic protein bim in epithelial cells
Virus Research, 2016Co-Authors: Maria Setas Lopes Pontes, Hans Nauwynck, Bruno Verhasselt, Cliff Van Waesberghe, Herman FavoreelAbstract:ERK1/2 (Extracellular signal Regulated Kinase 1/2) signaling is a key cellular signaling axis controlling many cellular events, including cell survival. Activation of ERK 1/2 may trigger an anti-apoptotic response, and different Viruses have been shown to benefit from this process. We have described recently that the viral glycoprotein gE mediates Pseudorabies Virus (PRV)-induced activation of ERK 1/2 in T lymphocytes. In the present study, we report that PRV gE-mediated ERK 1/2 phosphorylation also occurs in epithelial cells and that in these cells, gE-mediated ERK 1/2 signaling is associated with degradation of the pro-apoptotic protein Bim. Our results for the first time link the viral glycoprotein gE, an important alphaherpesVirus virulence factor, with the apoptotic signaling pathway.
Hans Nauwynck - One of the best experts on this subject based on the ideXlab platform.
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Pseudorabies Virus glycoprotein ge triggers erk1 2 phosphorylation and degradation of the pro apoptotic protein bim in epithelial cells
Virus Research, 2016Co-Authors: Maria Setas Lopes Pontes, Hans Nauwynck, Cliff Van Waesberghe, Bruno Verhasselt, Herman FavoreelAbstract:ERK1/2 (Extracellular signal Regulated Kinase 1/2) signaling is a key cellular signaling axis controlling many cellular events, including cell survival. Activation of ERK 1/2 may trigger an anti-apoptotic response, and different Viruses have been shown to benefit from this process. We have described recently that the viral glycoprotein gE mediates Pseudorabies Virus (PRV)-induced activation of ERK 1/2 in T lymphocytes. In the present study, we report that PRV gE-mediated ERK 1/2 phosphorylation also occurs in epithelial cells and that in these cells, gE-mediated ERK 1/2 signaling is associated with degradation of the pro-apoptotic protein Bim. Our results for the first time link the viral glycoprotein gE, an important alphaherpesVirus virulence factor, with the apoptotic signaling pathway.
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Pseudorabies Virus glycoprotein ge triggers erk1 2 phosphorylation and degradation of the pro apoptotic protein bim in epithelial cells
Virus Research, 2016Co-Authors: Maria Setas Lopes Pontes, Hans Nauwynck, Bruno Verhasselt, Cliff Van Waesberghe, Herman FavoreelAbstract:ERK1/2 (Extracellular signal Regulated Kinase 1/2) signaling is a key cellular signaling axis controlling many cellular events, including cell survival. Activation of ERK 1/2 may trigger an anti-apoptotic response, and different Viruses have been shown to benefit from this process. We have described recently that the viral glycoprotein gE mediates Pseudorabies Virus (PRV)-induced activation of ERK 1/2 in T lymphocytes. In the present study, we report that PRV gE-mediated ERK 1/2 phosphorylation also occurs in epithelial cells and that in these cells, gE-mediated ERK 1/2 signaling is associated with degradation of the pro-apoptotic protein Bim. Our results for the first time link the viral glycoprotein gE, an important alphaherpesVirus virulence factor, with the apoptotic signaling pathway.
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Genome Sequences of Two Pseudorabies Virus Strains Isolated in Greece
Genome announcements, 2016Co-Authors: Konstantinos V. Papageorgiou, Andrew J. Davison, Hans Nauwynck, Nicolás M. Suárez, Gavin S. Wilkie, George Filioussis, Nikolaos Papaioannou, Spyridon K. KritasAbstract:Pseudorabies Virus (species Suid herpesVirus 1) belongs to the genus VaricelloVirus, subfamily Alphaherpesvirinae, family Herpesviridae, and is the causative agent of an acute and frequently fatal disease that affects mainly pigs. Here, we report the genome sequences of two strains of this Virus isolated in Greece in 2010.
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a trypsin like serine protease is involved in Pseudorabies Virus invasion through the basement membrane barrier of porcine nasal respiratory mucosa
Veterinary Research, 2011Co-Authors: Sarah Glorieux, Herman Favoreel, Lennert Steukers, Annelies Vandekerckhove, Hans NauwynckAbstract:Several alphaherpesViruses breach the basement membrane during mucosal invasion. In the present study, the role of proteases in this process was examined. The serine protease-specific inhibitor AEBSF inhibited penetration of the basement membrane by the porcine alphaherpesVirus Pseudorabies Virus (PRV) by 88.1% without affecting lateral spread. Inhibitors of aspartic-, cysteine-, and metalloproteases did not inhibit viral penetration of the basement membrane. Further analysis using the Soybean Type I-S trypsin inhibitor for the serine protease subcategory of trypsin-like serine proteases resulted in a 96.9% reduction in plaque depth underneath the basement membrane. These data reveal a role of a trypsin-like serine protease in PRV penetration of the basement membrane.
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Cell biological and molecular characteristics of Pseudorabies Virus infections in cell cultures and in pigs with emphasis on the respiratory tract
Veterinary Research, 2007Co-Authors: Hans Nauwynck, Sarah Glorieux, Herman Favoreel, Maurice PensaertAbstract:In the present review, several cell biological and molecular aspects of Virus-cell and Virus-host (pig) interactions are reviewed for Pseudorabies (Aujeszky's disease) Virus. Concerning the Virus-cell interactions, the complex cascade of events in the Virus replication cycle is given together with the different mechanisms of cell-to-cell spread. The pathogenesis of Pseudorabies Virus infections in pigs is concentrated on the sequence of events in the respiratory tract. Finally, a short overview is given on the control of the disease and eradication of the Virus by the combination of marker vaccines and discriminating ELISA.
Dora Tombacz - One of the best experts on this subject based on the ideXlab platform.
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an integrated sequencing approach for updating the Pseudorabies Virus transcriptome
Pathogenetics, 2021Co-Authors: Gabor Torma, Dora Tombacz, Zsolt Csabai, Michael Snyder, Daniel Gobhardter, Zoltan Deim, Zsolt BoldogkőiAbstract: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.
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transcriptome wide survey of Pseudorabies Virus using next and third generation sequencing platforms
Scientific Data, 2018Co-Authors: Dora Tombacz, Donald Sharon, Michael Snyder, Attila Szűcs, Norbert Moldovan, Zsolt BoldogkőiAbstract:Pseudorabies Virus (PRV) is an alphaherpesVirus of swine. PRV has a large double-stranded DNA genome and, as the latest investigations have revealed, a very complex transcriptome. Here, we present a large RNA-Seq dataset, derived from both short- and long-read sequencing. The dataset contains 1.3 million 100 bp paired-end reads that were obtained from the Illumina random-primed libraries, as well as 10 million 50 bp single-end reads generated by the Illumina polyA-seq. The Pacific Biosciences RSII non-amplified method yielded 57,021 reads of inserts (ROIs) aligned to the viral genome, the amplified method resulted in 158,396 PRV-specific ROIs, while we obtained 12,555 ROIs using the Sequel platform. The Oxford Nanopore's MinION device generated 44,006 reads using their regular cDNA-sequencing method, whereas 29,832 and 120,394 reads were produced by using the direct RNA-sequencing and the Cap-selection protocols, respectively. The raw reads were aligned to the PRV reference genome (KJ717942.1). Our provided dataset can be used to compare different sequencing approaches, library preparation methods, as well as for validation and testing bioinformatic pipelines.
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multi platform sequencing approach reveals a novel transcriptome profile in Pseudorabies Virus
Frontiers in Microbiology, 2018Co-Authors: Norbert Moldovan, Dora Tombacz, Zsolt Csabai, Michael Snyder, Attila Szűcs, Zsolt BoldogkőiAbstract:Third-generation sequencing is an emerging technology that is capable of solving several problems that earlier approaches were not able to, including the identification of transcripts isoforms and overlapping transcripts. In this study, we used long-read sequencing for the analysis of Pseudorabies Virus (PRV) transcriptome, including Oxford Nanopore Technologies MinION, PacBio RS-II, and Illumina HiScanSQ platforms. We also used data from our previous short-read and long-read sequencing studies for the comparison of the results and in order to confirm the obtained data. Our investigations identified 19 formerly unknown putative protein-coding genes, all of which are 5’ truncated forms of earlier annotated longer PRV genes. Additionally, we detected 19 non-coding RNAs, including 5’ and 3’ truncated transcripts without in-frame ORFs, antisense RNAs, as well as RNA molecules encoded by those parts of the viral genome where no transcription had been detected before. This study has also led to the identification of three complex transcripts and fifty distinct length isoforms, including transcription start and end variants. We also detected 121 novel transcript overlaps, and two transcripts that overlap the replication origins of PRV. Furthermore, in silico analysis revealed 145 upstream ORFs, many of which are located on the longer 5’ isoforms of the transcripts.
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characterization of novel transcripts in Pseudorabies Virus
Viruses, 2015Co-Authors: Dora Tombacz, Donald Sharon, Peter Olah, Zsolt Csabai, Michael Snyder, Zoltan Havelda, Zsolt BoldogkőiAbstract:In this study we identified two 3'-coterminal RNA molecules in the Pseudorabies Virus. The highly abundant short transcript (CTO-S) proved to be encoded between the ul21 and ul22 genes in close vicinity of the replication origin (OriL) of the Virus. The less abundant long RNA molecule (CTO-L) is a transcriptional readthrough product of the ul21 gene and overlaps OriL. These polyadenylated RNAs were characterized by ascertaining their nucleotide sequences with the Illumina HiScanSQ and Pacific Biosciences Real-Time (PacBio RSII) sequencing platforms and by analyzing their transcription kinetics through use of multi-time-point Real-Time RT-PCR and the PacBio RSII system. It emerged that transcription of the CTOs is fully dependent on the viral transactivator protein IE180 and CTO-S is not a microRNA precursor. We propose an interaction between the transcription and replication machineries at this genomic location, which might play an important role in the regulation of DNA synthesis.
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strain kaplan of Pseudorabies Virus genome sequenced by pacbio single molecule real time sequencing technology
Genome Announcements, 2014Co-Authors: Dora Tombacz, Donald Sharon, Peter Olah, Zsolt Csabai, Michael Snyder, Zsolt BoldogkőiAbstract:Pseudorabies Virus (PRV) is a neurotropic herpesVirus that causes Aujeszky's disease in pigs. PRV strains are widely used as transsynaptic tracers for mapping neural circuits. We present here the complete and fully annotated genome sequence of strain Kaplan of PRV, determined by Pacific Biosciences RSII long-read sequencing technology.
