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Open AccessArticle

Amino Acid at Position 166 of NS2A in Japanese Encephalitis Virus (JEV) Is Associated with In Vitro Growth Characteristics of JEV

Department of Virology I, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku, Tokyo 162-8640, Japan
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Author to whom correspondence should be addressed.
Viruses 2020, 12(7), 709; https://doi.org/10.3390/v12070709
Received: 2 June 2020 / Revised: 26 June 2020 / Accepted: 29 June 2020 / Published: 30 June 2020
(This article belongs to the Section Animal Viruses)
We previously showed that the growth ability of the Japanese encephalitis virus (JEV) genotype V (GV) strain Muar is clearly lower than that of the genotype I (GI) JEV strain Mie/41/2002 in murine neuroblastoma cells. Here, we sought to identify the region in GV JEV that is involved in its low growth potential in cultured cells. An intertypic virus containing the NS1-3 region of Muar in the Mie/41/2002 backbone (NS1-3Muar) exhibited a markedly diminished growth ability in murine neuroblastoma cells. Moreover, the growth rate of a Muar NS2A-bearing intertypic virus (NS2AMuar) was also similar to that of Muar in these cells, indicating that NS2A of Muar is one of the regions responsible for the Muar-specific growth ability in murine neuroblastoma cells. Sequencing analysis of murine neuroblastoma Neuro-2a cell-adapted NS1-3Muar virus clones revealed that His-to-Tyr mutation at position 166 of NS2A (NS2A166) could rescue the low replication ability of NS1-3Muar in Neuro-2a cells. Notably, a virus harboring a Tyr-to-His substitution at NS2A166 (NS2AY166H) showed a decreased growth ability relative to that of the parental virus Mie/41/2002, whereas an NS2AMuar-based mutant virus, NS2AMuar-H166Y, showed a higher growth ability than NS2AMuar in Neuro-2a cells. Thus, these results indicate that the NS2A166 amino acid in JEV is critical for the growth and tissue tropism of JEV in vitro. View Full-Text
Keywords: Japanese encephalitis virus; genotype V; NS2A; reverse genetics Japanese encephalitis virus; genotype V; NS2A; reverse genetics
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Tajima, S.; Taniguchi, S.; Nakayama, E.; Maeki, T.; Inagaki, T.; Lim, C.-K.; Saijo, M. Amino Acid at Position 166 of NS2A in Japanese Encephalitis Virus (JEV) Is Associated with In Vitro Growth Characteristics of JEV. Viruses 2020, 12, 709.

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