Functional characterization of EUL47 in productive replication, morphogenesis and infectivity of equine herpesvirus 1.
Abstract: EUL47 is a major component of the tegument of equine herpesvirus 1 (EHV-1). To determine its function, we used Red/ET cloning to delete its gene (gene 13) from EHV-1 strain Ab4p inserted into a bacterial artificial chromosome (BAC), yielding Ab4pattBΔ13. We also examined the reverted virus (Ab4pattB13R). Ab4pattBΔ13 replicated in rabbit kidney (RK)-13 cells, indicating that ORF13 is dispensable for virus replication in cell culture. Its intracellular and extracellular titers were about 10- and 100-fold lower than those of the revertant and parent strains, respectively. In addition, the plaque size was half the plaque sizes of the other two strains. The particle-to-plaque forming unit ratio of Ab4pattBΔ13 was 21-fold greater than the ratios of the revertant and parent strains. No enveloped virions were detected in the cytoplasm of Ab4pattBΔ13-infected cells by transmission electron microscopy. In hamster, Ab4pattBΔ13 caused clinical signs and weight loss after only 1 day, but induced less severe neurological signs than did the revertant and parent strains. These results indicate that EUL47 is structurally required for normal virus replication, viral morphogenesis and viral infectivity, and that loss of EUL47 moderately attenuates the neuropathogenicity of EHV-1 in the hamster model.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication Date: 2011-10-26 PubMed ID: 22056848DOI: 10.1016/j.virusres.2011.10.021Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article observes the role of EUL47 in the development and severity of equine herpesvirus 1 (EHV-1). The findings suggest that EUL47 is structurally important for virus replication, development, and infectivity and that its absence lessens the severity of neuropathogenicity.
Objectives and Methods
- The aim of the study was to comprehend the function of EUL47, a major part of the equine herpesvirus 1 (EHV-1) structure.
- The researchers utilized a method known as Red/ET cloning to eliminate the EUL47 gene (gene 13) from the EHV-1 strain Ab4p, which was inserted into a bacterial artificial chromosome (BAC) to acquire Ab4pattBΔ13.
- The researchers also analyzed the reverted virus, Ab4pattB13R, to establish a comparison.
- Replication studies were conducted in rabbit kidney (RK)-13 cells to gauge whether the ORF13 is essential for viral replication in cell culture.
Findings
- Ab4pattBΔ13 was capable of replication in rabbit kidney (RK)-13 cells, suggesting that ORF13 is not necessary for virus replication in a cell culture environment.
- The intracellular and extracellular titers of Ab4pattBΔ13 were observed to be notably lower than those of the locale variant and parent strain, respectively.
- In addition, the size of the virus clusters (plaques) of Ab4pattBΔ13 was about half compared to the revertant and parent strains.
- The ratio of virus particles to plaque-forming units in Ab4pattBΔ13 was 21 times greater compared to the revertant and parent variants, indicating a likely reduced efficacy in generating infection.
- Furthermore, no enveloped virions were detected in the cytoplasm of cells infected by Ab4pattBΔ13.
Implications
- The study suggests that EUL47 is necessary for normal replication, form, and infectivity of the equine herpesvirus and that the loss of EUL47 mitigates the virus’s neuropathogenic impacts.
- The results could potentially inform future targeted treatments and vaccine strategies against EHV-1.
Cite This Article
APA
Yu MH, Kasem S, Yoshizaki N, Pagamjav O, Yamaguchi T, Ohya K, Fukushi H.
(2011).
Functional characterization of EUL47 in productive replication, morphogenesis and infectivity of equine herpesvirus 1.
Virus Res, 163(1), 310-319.
https://doi.org/10.1016/j.virusres.2011.10.021 Publication
Researcher Affiliations
- Department of Applied Veterinary Sciences, United Graduated School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
MeSH Terms
- Animals
- Cell Line
- Cricetinae
- Disease Models, Animal
- Gene Deletion
- Herpesviridae Infections / pathology
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / growth & development
- Herpesvirus 1, Equid / pathogenicity
- Herpesvirus 1, Equid / physiology
- Herpesvirus 1, Equid / ultrastructure
- Mesocricetus
- Microscopy, Electron, Transmission
- Rabbits
- Viral Load
- Viral Plaque Assay
- Viral Structural Proteins / genetics
- Viral Structural Proteins / metabolism
- Virion / ultrastructure
- Virulence
- Virus Replication
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