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Nucleotide at position 66 of NS2A in Japanese encephalitis virus is associated with the virulence and proliferation of virus

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Abstract

Most wild strains of Japanese encephalitis virus (JEV) produce NS1’ protein, which plays an important role in viral infection and immune escape. The G66A nucleotide mutation in NS2A gene of the wild strain SA14 prevented the ribosomal frameshift that prevented the production of NS1’ protein, thus reduced the virulence. In this study, the 66th nucleotide of the NS2A gene of SA14 was mutated into A, U or C, respectively. Both the G66U and G66C mutations cause the E22D mutation of the NS2A protein. Subsequently, the expression of NS1’ protein, plaque size, replication ability, and virulence to mice of the three mutant strains were examined. The results showed that the three mutant viruses could not express NS1’ protein, and their proliferation ability in nerve cells and virulence to mice were significantly reduced. In addition, the SA14(G66C) was less virulent than the other two mutated viruses. Our results indicate that only when G is the 66th nucleotide of NS2A, the JEV can produce NS1’ protein, which affects the virulence.

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Funding

This work was supported by the Research fund of North Sichuan Medical College (No. CBY22-ZDA06).

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Authors and Affiliations

  1. Department of Pathogenic Biology, School of Basic Medical Sciences and Forensic Medicine, North Sichuan Medical College, Nanchong, 637100, China

    Ning Tan, Chen Chen, Rong Huang, Zhuang Zhu, Kui Xu, Xiaoyao Yang, Jian Yang & Lei Yuan

  2. Department of Laboratory Medicine, Jintang First People’s Hospital, West China Hospital Sichuan University JinTang Hospital, Chengdu, 610400, China

    Yang Ren

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  1. Ning Tan
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Contributions

LY and NT designed the study, analyzed the data and wrote the manuscript. CC, YR and RH cultured viruses, collected and tested the samples and analyzed the data. ZZ, KX and XY carried out the animal experiment. JY and LY reviewed and edited the manuscript. All authors approved the final manuscript.

Corresponding author

Correspondence to Lei Yuan.

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Conflicts of interest

The authors declared that they have no conflicts of interest to this work.

Ethical approval

All procedures performed in this study involving animals were approved by the Institutional Animal Care and Use Committee of North Sichuan Medical College (approval number AE2022-068), Nanchong city, China and followed National Institutes of Health guidelines.

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Edited by Sassan Asgari.

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Tan, N., Chen, C., Ren, Y. et al. Nucleotide at position 66 of NS2A in Japanese encephalitis virus is associated with the virulence and proliferation of virus. Virus Genes 60, 9–17 (2024). https://doi.org/10.1007/s11262-023-02036-5

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  • DOI: https://doi.org/10.1007/s11262-023-02036-5

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