Picornavirus VP3 protein induces autophagy through the TP53-BAD-BAX axis to promote viral replication

ABSTRACT

Macroautophagy/autophagy and apoptosis are pivotal interconnected host cell responses to viral infection, including picornaviruses. Here, the VP3 proteins of picornaviruses were determined to trigger autophagy, with the autophagic flux being triggered by the TP53-BAD-BAX axis. Using foot-and-mouth disease virus (FMDV) as a model system, we unraveled a novel mechanism of how picornavirus hijacks autophagy to bolster viral replication and enhance pathogenesis. FMDV infection induced both autophagy and apoptosis in vivo and in vitro. FMDV VP3 protein facilitated the phosphorylation and translocation of TP53 from the nucleus into the mitochondria, resulting in BAD-mediated apoptosis and BECN1-mediated autophagy. The amino acid Gly129 in VP3 is essential for its interaction with TP53, and crucial for induction of autophagy and apoptosis. VP3-induced autophagy and apoptosis are both essential for FMDV replication, while, autophagy plays a more important role in VP3-mediated pathogenesis. Mutation of Gly129 to Ala129 in VP3 abrogated the autophagic regulatory function of VP3, which significantly decreased the viral replication and pathogenesis of FMDV. This suggested that VP3-induced autophagy benefits viral replication and pathogenesis. Importantly, this Gly is conserved and showed a common function in various picornaviruses. This study provides insight for developing broad-spectrum antivirals and genetic engineering attenuated vaccines against picornaviruses.

Abbreviations: 3-MA, 3-methyladenine; ATG, autophagy related; BAD, BCL2 associated agonist of cell death; BAK1, BCL2 antagonist/killer 1; BAX, BCL2 associated X, apoptosis regulator; BBC3/PUMA, BCL2 binding component 3; BCL2, BCL2 apoptosis regulator; BID, BH3 interacting domain death agonist; BIP-V5, BAX inhibitor peptide V5; CFLAR/FLIP, CASP8 and FADD like apoptosis regulator; CPE, cytopathic effects; CQ, chloroquine; CV, coxsackievirus; DAPK, death associated protein kinase; DRAM, DNA damage regulated autophagy modulator; EV71, enterovirus 71; FMDV, foot-and-mouth disease virus; HAV, hepatitis A virus; KD, knockdown; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; MOI, multiplicity of infection; MTOR, mechanistic target of rapamycin kinase; PML, promyelocytic leukemia; PV, poliovirus; SVA, Seneca Valley virus; TCID50, 50% tissue culture infectious doses; TOR, target of rapamycin. TP53/p53, tumor protein p53; WCL, whole-cell lysate.

KEYWORDS:

• Autophagy
• picornavirus
• replication
• TP53-BAD-BAX axis
• VP3

Acknowledgements

We thank the Facility Center Department and National Foot-and-Mouth Disease Reference Laboratory (ABSL-3), Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences and Analysis and Test Group, Center for Technical Development and Analysis Service, Institute of Modern Physics, for helpful experimental facilities and analysis support. This work was supported by grants from the National Key R&D Program of China (2021YFD1800300), Key Project of National Natural Sciences Foundation of China (32302894, 32330107), the Fundamental Research Funds for the Central Universities, the Key Technologies R&D Program of Gansu Province (23JRRA1515), the Open Competition Program of Top Ten Critical Priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province (2023SDZG02), the Project of National Center of Technology Innovation for Pigs (NCTIP-XD/C03), and the Earmarked Fund for CARS-35 and CARS-39-13,the Fundamental Research Funds for Innovation Team of Gansu Province (23JRRA546).

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

Data from this study are available from the Lead Contact, Dr. Haixue Zheng ([email protected]), upon reasonable request.

Supplemental data

Supplemental data for this article can be accessed online at https://doi.org/10.1080/15548627.2024.2350270.

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

The work was supported by the National Key Research and Development Program of China [2021YFD1800300].