Unique Localization of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Infected Cells

2016 ◽  
Vol 6 (1) ◽  
pp. 914-921 ◽  
Author(s):  
Yuto Suda ◽  
Daisuke Yamane ◽  
Muhammad Atif Zahoor ◽  
Yassir Mahgoub Mohamed ◽  
Shin Murakami ◽  
...  
Keyword(s):  
Viral Replication ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Replication Complex ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Infected Cells ◽  
Viral Replication Complex ◽  
Viral Diarrhea Virus ◽  
Ns4b Protein

Bovine viral diarrhea virus (BVDV), an important pathogen infecting ruminants, has 2 biotypes: cytopathic (cp) and noncytopathic (ncp), which are related to the onset of disease. The viral replication complex is composed of viral non-structural (NS) proteins, raising the possibility that NS protein(s) play a role in viral biotypes. To gain insight into this possible role, we analysed the intracellular localization of each NS protein in both cp and ncp virus-infected cells, and found that NS4B protein, a possible anchor protein of the viral replication complex, showed a unique dotted localization pattern that markedly merged with an endoplasmic reticulum marker, unlike other NS proteins, although there was no difference in the localization of NS4B protein between the 2 biotypes. 

scholarly journals Inhibition of Beta Interferon Transcription by Noncytopathogenic Bovine Viral Diarrhea Virus Is through an Interferon Regulatory Factor 3-Dependent Mechanism

2002 ◽  
Vol 76 (18) ◽  
pp. 8979-8988 ◽  
Author(s):  
Susan J. Baigent ◽  
Gang Zhang ◽  
Martin D. Fray ◽  
Helen Flick-Smith ◽  
Stephen Goodbourn ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Bovine Viral Diarrhea Virus ◽  
Interferon Regulatory Factor ◽  
Bovine Viral Diarrhea ◽  
Regulatory Factor ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Nuclear Extracts ◽  
Infected Cells ◽  
Viral Diarrhea Virus

ABSTRACT The induction and inhibition of the interferon (IFN) response and apoptosis by bovine viral diarrhea virus (BVDV) has been examined. Here we show that prior infection of cells by noncytopathogenic BVDV (ncp BVDV) fails to block transcriptional responses to alpha/beta IFN. In contrast, ncp BVDV-infected cells fail to produce IFN-α/β or MxA in response to double-stranded RNA (dsRNA) or infection with a heterologous virus (Semliki Forest virus [SFV]). ncp BVDV preinfection is unable to block cp BVDV- or SFV-induced apoptosis. The effects of ncp BVDV infection on the transcription factors controlling the IFN-β induction pathway have been analyzed. The transcription factor NF-κB was not activated following ncp BVDV infection, but ncp BVDV infection was not able to block the activation of NF-κB by either SFV or tumor necrosis factor alpha. Furthermore, ncp BVDV infection did not result in the activation of stress kinases (JNK1 and JNK2) or the phosphorylation of transcription factors ATF-2 and c-Jun; again, ncp BVDV infection was not able to block their activation by SFV. Interferon regulatory factor 3 (IRF-3) was shown to be translocated to the nuclei of infected cells in response to ncp BVDV, although DNA-binding of IRF-3 was not seen in nuclear extracts. In contrast, an IRF-3-DNA complex was observed in nuclear extracts from cells infected with SFV, but the appearance of this complex was blocked when cells were previously exposed to ncp BVDV. We conclude that the inhibition of IFN induction by this pestivirus involves a block to IRF-3 function, and we speculate that this may be a key characteristic for the survival of pestiviruses in nature.

scholarly journals Double-immunolabeling systems for phenotyping of immune cells harboring bovine viral diarrhea virus.

1987 ◽  
Vol 35 (6) ◽  
pp. 627-633 ◽  
Author(s):  
H Bielefeldt Ohmann
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Simultaneous Detection ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Tissue Sections ◽  
Specificity And Sensitivity ◽  
Infected Cells ◽  
Viral Diarrhea Virus ◽  
Double Immunolabeling

Optimal staining conditions were defined for simultaneous detection of bovine viral diarrhea virus (BVDV) and mononuclear leukocyte surface antigens in tissue sections and cytospins. Because of the extreme lability of the virus antigens and the variable stability of the epitopes on the cell differentiation antigens, cryopreservation had to be used. This method gives slightly sub-optimal preservation of morphology. However, the specificity and sensitivity of the immunolabeling ensured reliable identification of the double-labeled cells, i.e., the phenotypic identification of virus-infected cells within the immune system.

scholarly journals Establishment of a Subgenomic Replicon for Bovine Viral Diarrhea Virus in Huh-7 Cells and Modulation of Interferon-Regulated Factor 3-Mediated Antiviral Response

2005 ◽  
Vol 79 (5) ◽  
pp. 2788-2796 ◽  
Author(s):  
Nigel Horscroft ◽  
Dan Bellows ◽  
Israrul Ansari ◽  
Vicky C. H. Lai ◽  
Shannon Dempsey ◽  
...  
Keyword(s):  
Viral Replication ◽  
Bovine Viral Diarrhea Virus ◽  
Sendai Virus ◽  
Rna Replication ◽  
Bovine Viral Diarrhea ◽  
Subgenomic Replicon ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Amino Terminal ◽  
Viral Diarrhea Virus

ABSTRACT We describe the development of a selectable, bi-cistronic subgenomic replicon for bovine viral diarrhea virus (BVDV) in Huh-7 cells, similar to that established for hepatitis C virus (HCV). The selection marker and reporter (Luc-Ubi-Neo) in the BVDV replicon was fused with the amino-terminal protease Npro, and expression of the nonstructural proteins (NS3 to NS5B) was driven by an encephalomyocarditis virus internal ribosome entry site. This BVDV replicon allows us to compare RNA replication of these two related viruses in a similar cellular background and to identify antiviral molecules specific for HCV RNA replication. The BVDV replicon showed similar sensitivity as the HCV replicon to interferons (alpha, beta, and gamma) and 2′-β-C-methyl ribonucleoside inhibitors. Known nonnucleoside inhibitor molecules specific for either HCV or BVDV can be easily distinguished by using the parallel replicon systems. The HCV replicon has been shown to block, via the NS3/4A serine protease, Sendai virus-induced activation of interferon regulatory factor 3 (IRF-3), a key antiviral signaling molecule. Similar suppression of IRF-3-mediated responses was also observed with the Huh-7-BVDV replicon but was independent of NS3/4A protease activity. Instead, the amino-terminal cysteine protease Npro of BVDV appears to be, at least partly, responsible for suppressing IRF-3 activation induced by Sendai virus infection. This result suggests that different viruses, including those closely related, may have developed unique mechanisms for evading host antiviral responses. The parallel BVDV and HCV replicon systems provide robust counterscreens to distinguish viral specificity of small-molecule inhibitors of viral replication and to study the interactions of the viral replication machinery with the host cell innate immune system.

scholarly journals Essential and Nonessential Elements in the 3′ Nontranslated Region of Bovine Viral Diarrhea Virus

2005 ◽  
Vol 79 (14) ◽  
pp. 9119-9127 ◽  
Author(s):  
Alexander Pankraz ◽  
Heinz-Jürgen Thiel ◽  
Paul Becher
Keyword(s):  
Viral Replication ◽  
Bovine Viral Diarrhea Virus ◽  
Host Cells ◽  
Close Relative ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Nontranslated Region ◽  
Viral Diarrhea Virus ◽  
Efficient Viral Replication

ABSTRACT The 3′ nontranslated region (NTR) of the pestivirus Bovine viral diarrhea virus (BVDV), a close relative of human Hepatitis C virus, consists of three stem-loops which are separated by two single-stranded regions. As in other positive-stranded RNA viruses, the 3′ NTR of pestiviruses is involved in crucial processes of the viral life cycle. While several studies characterized cis-acting elements within the 3′ NTR of a BVDV replicon, there are no studies addressing the significance of these elements in the context of a replicating virus. To examine the functional importance of 3′ NTR elements, a set of 4-base deletions and deletions of each of the three stem-loops were introduced into an infectious BVDV cDNA clone. Emerging mutant viruses were characterized with regard to plaque phenotype, growth kinetics, and synthesis of viral RNA. The results indicated that presence of stem-loop (SL) I and the 3′-terminal part of the single-stranded region between stem-loops I and II are indispensable for pestiviral replication. In contrast, deletions within SL II and SL III as well as absence of either SL II or SL III still allowed efficient viral replication; however, a mutant RNA lacking both SL II and SL III was not infectious. The results of this study provide a detailed map of the essential and nonessential elements within the 3′ NTR of BVDV and contribute to our understanding of sequence and structural elements important for efficient viral replication of pestiviruses in natural host cells.

scholarly journals Gypenoside Inhibits Bovine Viral Diarrhea Virus Replication by Interfering with Viral Attachment and Internalization and Activating Apoptosis of Infected Cells

Viruses ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1810
Author(s):  
Guanghui Yang ◽  
Jialu Zhang ◽  
Shenghua Wang ◽  
Jun Wang ◽  
Jing Wang ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Host Cells ◽  
Bovine Viral Diarrhea ◽  
Tunel Staining ◽  
Viral Attachment ◽  
Innate Defense ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Infected Cells ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) causes a severe threat to the cattle industry due to ineffective control measures. Gypenoside is the primary component of Gynostemma pentaphyllum, which has potential medicinal value and has been widely applied as a food additive and herbal supplement. However, little is known about the antiviral effects of gypenoside. The present study aimed to explore the antiviral activities of gypenoside against BVDV infection. The inhibitory activity of gypenoside against BVDV was assessed by using virus titration and performing Western blotting, quantitative reverse transcription PCR (RT-qPCR), and immunofluorescence assays in MDBK cells. We found that gypenoside exhibited high anti-BVDV activity by interfering with the viral attachment to and internalization in cells. The study showed that BVDV infection inhibits apoptosis of infected cells from escaping the innate defense of host cells. Our data further demonstrated that gypenoside inhibited BVDV infection by electively activating the apoptosis of BVDV-infected cells for execution, as evidenced by the regulation of the expression of the apoptosis-related protein, promotion of caspase-3 activation, and display of positive TUNEL staining; no toxicity was observed in non-infected cells. Collectively, the data identified that gypenoside exerts an anti-BVDV-infection role by inhibiting viral attachment and internalization and selectively purging virally infected cells. Therefore, our study will contribute to the development of a novel prophylactic and therapeutic strategy against BVDV infection.

Electron Microscopic study of two strains of bovine viral diarrhea virus (BVDV)

1989 ◽  
Vol 47 ◽  
pp. 1030-1031
Author(s):  
R. Alain ◽  
S. Cardin ◽  
L. Berthiaume ◽  
J. Lecomte
Keyword(s):  
Endoplasmic Reticulum ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Icosahedral Symmetry ◽  
Electron Microscopic ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Wide Range ◽  
Infected Cells ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle throughout the world, causing a wide range of clinical syndromes. Virion is spherical, 40-70 nm in diameter, with an envelope tightly applied to a spherical nucleocapsid 25-35 nm in diameter with icosahedral symmetry (figure 1 and 2) (Matthews, 1979) . It is presently included in the Togavlridae family, genus Pneumovirus but Collett et al. (1988) have proposed to group this genus in the Flaviviridae family. In the present study, two strains of BVDV were compared by electron microcospy ultrathin sections: one cytopathogenic, NADL strain, and the other non-cytopathogenic, NY-1 strain. These were Inoculated on Madney-Darby Bovine Kidney (MDBK) cells to a multiplicity of infection (m.o.i) between 3,0 and 5,0. Control and virus infected cells were fixed with glutaraldehyde (2,5% in cacodylate buffer) at 24 , 48, 72 and 96 h following incubation at 37°C. Cells were postfixed with osmium tetroxide and embedded in Vestopal.The NADL-CP strain infected cells showed vacuolization of the endoplasmic reticulum, ribosomal activity in the cytoplasm and appearance of virus particles 48h after infection (figure 3). At 72h, an expensive vacuolization was apparent (figure 4). Maximum viral production was seen 96 h after infection (figure 5). From 24 to 72 h post infection, the NY-l-NCP strain of BVD infected cells showed normal vacuolization like in control cells. They produced weak vacuolization with few particles, not apparent before 96h after infection (figure 6). Viral particles of 45-50 nm in endoplasmic reticulum vacuoles were in general homogenous in size while those in smooth membranes, probably of cellular origin, heterogenous in size.

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