scholarly journals The Npro product of classical swine fever virus interacts with IκBα, the NF-κB inhibitor

2008 ◽  
Vol 89 (8) ◽  
pp. 1881-1889 ◽  
Author(s):  
Virginie Doceul ◽  
Bryan Charleston ◽  
Helen Crooke ◽  
Elizabeth Reid ◽  
Penny P. Powell ◽  
...  
Keyword(s):  
Immune Response ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Host Cells ◽  
Nuclear Accumulation ◽  
Yeast Two Hybrid ◽  
Necrosis Factor Alpha ◽  
Induced Apoptosis ◽  
Two Hybrid

Classical swine fever virus (CSFV) belongs to the genus Pestivirus and is the causative agent of classical swine fever, a haemorrhagic disease of pigs. The virus replicates in host cells without activating interferon (IFN) production and has been reported to be an antagonist of double-stranded RNA-induced apoptosis. The N-terminal protease (Npro) of CSFV is responsible for this evasion of the host innate immune response. In order to identify cellular proteins that interact with the Npro product of CSFV, a yeast two-hybrid screen of a human library was carried out, which identified IκBα, the inhibitor of NF-κB, a transcription factor involved in the control of apoptosis, the immune response and IFN production. The Npro–IκBα interaction was confirmed using yeast two-hybrid analysis and additional co-precipitation assays. It was also shown that Npro localizes to both the cytoplasmic and nuclear compartments in stably transfected cells and in CSFV-infected cells. Following stimulation by tumour necrosis factor alpha, PK-15 cell lines expressing Npro exhibited transient nuclear accumulation of pIκBα, but no effect of CSFV infection on IκBα localization or NF-κB p65 activation was observed.

scholarly journals SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine

Viruses ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 421
Author(s):  
Elizabeth A. Vuono ◽  
Elizabeth Ramirez-Medina ◽  
Paul Azzinaro ◽  
Keith A. Berggren ◽  
Ayushi Rai ◽  
...  
Keyword(s):  
Protein Interaction ◽  
Classical Swine Fever Virus ◽  
Critical Role ◽  
Hybrid Approach ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Proximity Ligation Assay ◽  
Yeast Two Hybrid ◽  
Clinically Normal ◽  
Two Hybrid

E2 is the major structural glycoprotein of the classical swine fever virus (CSFV). E2 has been shown to be involved in important virus functions such as replication and virulence in swine. Using the yeast two-hybrid system, we previously identified several host proteins specifically interacting with CSFV E2. Here, we analyze the protein interaction of E2 with SERTA domain containing protein 1 (SERTAD1), a factor involved in the stimulation of the transcriptional activities of different host genes. We have confirmed that the interaction between these two proteins occurs in CSFV-infected swine cells by using a proximity ligation assay and confocal microscopy. Amino acid residues in the CSFV E2 protein that are responsible for mediating the interaction with SERTAD1 were mapped by a yeast two-hybrid approach using a randomly mutated E2 library. Using that information, a recombinant CSFV mutant (E2ΔSERTAD1v) that harbors substitutions in those residues mediating the protein-interaction with SERTAD1 was developed and used to study the role of the E2-SERTAD1 interaction in viral replication and virulence in swine. CSFV E2ΔSERTAD1v, when compared to the parental BICv, showed a clearly decreased ability to replicate in the SK6 swine cell line and a more severe replication defect in primary swine macrophage cultures. Importantly, 80% of animals infected with E2ΔSERTAD1v survived infection, remaining clinically normal during the 21-day observational period. This result would indicate that the ability of CSFV E2 to bind host SERTAD1 protein during infection plays a critical role in virus virulence.

Interactive cellular proteins related to classical swine fever virus non-structure protein 2 by yeast two-hybrid analysis

2012 ◽  
Vol 39 (12) ◽  
pp. 10515-10524 ◽  
Author(s):  
Kai Kang ◽  
Kangkang Guo ◽  
Qinhai Tang ◽  
Yanming Zhang ◽  
Jiang Wu ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Cellular Proteins ◽  
Yeast Two Hybrid ◽  
Hybrid Analysis ◽  
Two Hybrid

scholarly journals Swine Host Protein Coiled-Coil Domain-Containing 115 (CCDC115) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2 during Virus Replication

Viruses ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 388 ◽  
Author(s):  
Elizabeth A. Vuono ◽  
Elizabeth Ramirez-Medina ◽  
Keith Berggren ◽  
Ayushi Rai ◽  
Sarah Pruitt ◽  
...  
Keyword(s):  
Amino Acid ◽  
Virus Replication ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Host Protein ◽  
Yeast Two Hybrid ◽  
Host Proteins ◽  
E2 Protein ◽  
Two Hybrid

Interactions between the major structural glycoprotein E2 of classical swine fever virus (CSFV) with host proteins have been identified as important factors affecting virus replication and virulence. Previously, using the yeast two-hybrid system, we identified swine host proteins specifically interacting with CSFV E2. In this report, we use a proximity ligation assay to demonstrate that swine host protein CCDC115 interacts with E2 in CSFV-infected swine cells. Using a randomly mutated E2 library in the context of a yeast two-hybrid methodology, specific amino acid mutations in the CSFV E2 protein responsible for disrupting the interaction with CCDC115 were identified. A recombinant CSFV mutant (E2ΔCCDC115v) harboring amino acid changes disrupting the E2 protein interaction with CCDC115 was produced and used as a tool to assess the role of the E2–CCDC115 interaction in viral replication and virulence in swine. CSFV E2ΔCCDC115v showed a slightly decreased ability to replicate in the SK6 swine cell line and a greater replication defect in primary swine macrophage cultures. A decreased E2–CCDC115 interaction detected by PLA is observed in cells infected with E2ΔCCDC115v. Importantly, animals intranasally infected with 105 TCID50 of E2ΔCCDC115v experienced a significantly longer survival period when compared with those infected with the parental Brescia strain. This result would indicate that the ability of CSFV E2 to bind host CCDC115 protein during infection plays an important role in virus replication in swine macrophages and in virus virulence during the infection in domestic swine.

scholarly journals Analysis of Virus Population Profiles within Pigs Infected with Virulent Classical Swine Fever Viruses: Evidence for Bottlenecks in Transmission but Absence of Tissue-Specific Virus Variants

2020 ◽  
Vol 94 (19) ◽  
Author(s):  
Camille Melissa Johnston ◽  
Ulrik Fahnøe ◽  
Louise Lohse ◽  
Jens Bukh ◽  
Graham J. Belsham ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Population Change ◽  
Virulent Strain ◽  
Severe Disease ◽  
Clinical Signs ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Host Cells ◽  
Virus Population ◽  
Highly Virulent

ABSTRACT Classical swine fever virus (CSFV) contains a specific motif within the E2 glycoprotein that differs between strains of different virulence. In the highly virulent CSFV strain Koslov, this motif comprises residues S763/L764 in the polyprotein. However, L763/P764 represent the predominant alleles in published CSFV genomes. In this study, changes were introduced into the CSFV strain Koslov (here called vKos_SL) to generate modified CSFVs with substitutions at residues 763 and/or 764 (vKos_LL, vKos_SP, and vKos_LP). The properties of these mutant viruses, in comparison to those of vKos_SL, were determined in pigs. Each of the viruses was virulent and induced typical clinical signs of CSF, but the vKos_LP strain produced them significantly earlier. Full-length CSFV cDNA amplicons (12.3 kb) derived from sera of infected pigs were deep sequenced and cloned to reveal the individual haplotypes that contributed to the single-nucleotide polymorphism (SNP) profiles observed in the virus population. The SNP profiles for vKos_SL and vKos_LL displayed low-level heterogeneity across the entire genome, whereas vKos_SP and vKos_LP displayed limited diversity with a few high-frequency SNPs. This indicated that vKos_SL and vKos_LL exhibited a higher level of fitness in the host and more stability at the consensus level, whereas several consensus changes were observed in the vKos_SP and vKos_LP sequences, pointing to adaptation. For each virus, only a subset of the variants present within the virus inoculums were maintained in the infected pigs. No clear tissue-dependent quasispecies differentiation occurred within inoculated pigs; however, clear evidence for transmission bottlenecks to contact animals was observed, with subsequent loss of sequence diversity. IMPORTANCE The surface-exposed E2 protein of classical swine fever virus is required for its interaction with host cells. A short motif within this protein varies between strains of different virulence. The importance of two particular amino acid residues in determining the properties of a highly virulent strain of the virus has been analyzed. Each of the different viruses tested proved highly virulent, but one of them produced earlier, but not more severe, disease. By analyzing the virus genomes present within infected pigs, it was found that the viruses which replicated within inoculated animals were only a subset of those within the virus inoculum. Furthermore, following contact transmission, it was shown that a very restricted set of viruses had transferred between animals. There were no significant differences in the virus populations present in various tissues of the infected animals. These results indicate mechanisms of virus population change during transmission between animals.

scholarly journals In vitro infection with classical swine fever virus inhibits the transcription of immune response genes

2012 ◽  
Vol 9 (1) ◽  
pp. 175 ◽  
Author(s):  
Li Feng ◽  
Xiao-Quan Li ◽  
Xiao-ning Li ◽  
Jun Li ◽  
Xian-Ming Meng ◽  
...  
Keyword(s):  
Immune Response ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Immune Response Genes

Classical swine fever virus NS2 protein promotes interleukin-8 expression and inhibits MG132-induced apoptosis

Virus Genes ◽  
2011 ◽  
Vol 42 (3) ◽  
pp. 355-362 ◽  
Author(s):  
Qinghai Tang ◽  
Kangkang Guo ◽  
Kai Kang ◽  
Yanming Zhang ◽  
Lei He ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Interleukin 8 ◽  
Induced Apoptosis

40. Interactions between Classical swine fever virus and host cells

2002 ◽  
Vol 72 ◽  
pp. 14
Author(s):  
E. Bensaude ◽  
D. Sweetman ◽  
D.J. Paton ◽  
T.W. Drew ◽  
T. Wileman ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Host Cells

scholarly journals Different evolutionary patterns of classical swine fever virus envelope proteins

2016 ◽  
Vol 62 (3) ◽  
pp. 210-219 ◽  
Author(s):  
Yan Li ◽  
Zexiao Yang ◽  
Mingwang Zhang
Keyword(s):  
Classical Swine Fever Virus ◽  
Functional Divergence ◽  
Classical Swine Fever ◽  
Acid Sites ◽  
Envelope Proteins ◽  
Viral Envelope ◽  
Fever Virus ◽  
Host Cells ◽  
Virus Envelope ◽  
Evolutionary Patterns

Classical swine fever virus (CSFV) is the causative agent of classical swine fever, which is a highly contagious disease of the domestic pig as well as wild boar. The proteins Erns, E1, and E2 are components of the viral envelope membrane. They are also implicated in virus attachment and entry, replication, and (or) anti-immune response. Here, we studied the genetic variations of these envelope proteins in the evolution of CSFV. The results reveal that the envelope proteins underwent different evolutionary fates. In Ernsand E1, but not E2, a number of amino acid sites experienced functional divergence. Furthermore, the diversification in Ernsand E1 was generally episodic because the divergence-related changes of E1 only occurred with the separation of 2 major groups of CSFV and that of Ernstook place with the division of 1 major group. The major divergence-related sites of Ernsare located on one of the substrate-binding regions of the RNase domain and C-terminal extension. These functional domains have been reported to block activation of the innate immune system and attachment and entry into host cells, respectively. Our results may shed some light on the divergent roles of the envelope proteins.

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