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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Virology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1485667

Japanese encephalitis virus infection induces mitochondrial-mediated apoptosis through the pro-apoptotic protein BAX

Provisionally accepted
KE YANG KE YANG 1Xinran Li Xinran Li 1*Shuqing Yang Shuqing Yang 1*Yi Zheng Yi Zheng 1*Sanjie Cao Sanjie Cao 1Qigui YAN Qigui YAN 1Xiaobo Huang Xiaobo Huang 1Yiping Wen Yiping Wen 1QIN ZHAO QIN ZHAO 1Senyan Du Senyan Du 1Yifei Lang Yifei Lang 1Shan Zhao Shan Zhao 1Rui Wu Rui Wu 1,2*
  • 1 Sichuan Agricultural University, Ya'an, China
  • 2 Sichuan Science-observation Experimental Station of Veterinary Drugs and Veterinary Diagnostic Technology, Ministry of Agriculture, Chengdu, Sichuan Province, China

The final, formatted version of the article will be published soon.

    Abstract：Japanese encephalitis virus (JEV), a zoonotic flavivirus, is the primary cause of viral encephalitis in Asia. JEV induces apoptosis in a variety of cells. Nevertheless, the precise mechanism underlying apoptosis induced by JEV infection remains to be elucidated. Our previous studies showed that the pro-apoptosis gene BAX may have a role in JEV proliferation. In this study, we constructed a PK-15 cell line (BAX.KO) with BAX gene knockout by CRISPR/Cas9.BAX gene knockout effectively inhibits the proliferation of JEV, Viral protein levels decreased by 39.9%,whereas BAX overexpression has the opposite effect. We confirmed that JEV could induce the apoptosis of PK-15 by DAPI staining and Annexin V-FITC/PI staining.Furthermore, we found that the phosphorylation of P53 and the expression levels of BAX, NOXA, PUMA, and cleaved-caspase-3/9 were significantly upregulated after JEV infection. Moreover, we found that JEV infection not only caused mitochondrial damage, the release of mitochondrial cytochrome C (Cyt C), and the downregulation of the apoptosisinhibiting protein BCL-2 but also reduced the mitochondrial membrane potential (MOMP) and the accumulation of intracellular reactive oxygen species (ROS). These factors collectively encourage the activation of the mitochondrial apoptosis pathway. In contrast, BAX gene knockout significantly reduces the apoptotic changes caused by JEV infection. Treatment with the caspase3 inhibitor attenuated JEV-induced viral proliferation and release, and the viral protein levels of PK-15 and BAX.KO cells were reduced by 46% and 30%, respectively. In conclusion, this study clarified the molecular mechanism of JEV-induced apoptosis and provided a theoretical basis for revealing the pathogenic mechanism of JEV infection.

    Keywords: JEV, Mitochondrial apoptotic pathway, Bax, P53-BAX pathway, PK-15

    Received: 24 Aug 2024; Accepted: 27 Sep 2024.

    Copyright: © 2024 YANG, Li, Yang, Zheng, Cao, YAN, Huang, Wen, ZHAO, Du, Lang, Zhao and Wu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Xinran Li, Sichuan Agricultural University, Ya'an, China
    Shuqing Yang, Sichuan Agricultural University, Ya'an, China
    Yi Zheng, Sichuan Agricultural University, Ya'an, China
    Rui Wu, Sichuan Agricultural University, Ya'an, China

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