Functional characterization of EUL47 in productive replication, morphogenesis and infectivity of equine herpesvirus 1 |
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Authors: | Yu Mi Htay Htay Kasem Samy Yoshizaki Norio Pagamjav Ochir Yamaguchi Tsuyoshi Ohya Kenji Fukushi Hideto |
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Institution: | a Department of Applied Veterinary Sciences, United Graduated School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan b Department of Animal Science, Faculty of Applied Biological Sciences, Gifu 501-1193, Japan c The Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University, 4-101 Koyama Minami, Tottori 680-8550, Japan d Laboratory of Veterinary Microbiology, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan e Department of Virology, Faculty of Veterinary Medicine, kafr El Sheikh University, kafr El Sheikh 33516, Egypt |
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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. |
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Keywords: | Equine herpesvirus 1 EUL47-null mutant Pathogenicity Morphogenesis |
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