Regulation of host factor γ-H2AX level and location by enterovirus A71 for viral replication

ABSTRACT

Numerous viruses manipulate host factors for viral production. We demonstrated that human enterovirus A71 (EVA71), a primary causative agent for hand, foot, and mouth disease (HFMD), increased the level of the DNA damage response (DDR) marker γ-H2AX. DDR is primarily mediated by the ataxia telangiectasia mutated (ATM), ATM and Rad3-related (ATR), or DNA-dependent protein kinase (DNA-PK) pathways. Upregulation of γ-H2AX by EVA71 was dependent on the ATR but not the ATM or DNA-PK pathway. As a nuclear factor, there is no previous evidence of cytoplasmic distribution of γ-H2AX. However, the present findings demonstrated that EVA71 encouraged the localization of γ-H2AX to the cytoplasm. Of note, γ-H2AX formed a complex with structural protein VP3, non-structural protein 3D, and the viral genome. Treatment with an inhibitor or CRISPR/Cas9 technology to decrease or silence the expression of γ-H2AX decreased viral genome replication in host cells; this effect was accompanied by decreased viral protein expression and virions. In animal experiments, caffeine was used to inhibit DDR; the results revealed that caffeine protected neonatal mice from death after infection with EVA71, laying the foundation for new therapeutic applications of caffeine. More importantly, in children with HFMD, γ-H2AX was upregulated in peripheral blood lymphocytes. The consistent in vitro and in vivo data on γ-H2AX from this study suggested that caffeine or other inhibitors of DDR might be novel therapeutic agents for HFMD.

KEYWORDS:

• Enterovirus A71 (EVA71)
• γ-H2AX level
• γ-H2ax location
• DNA damage response
• viral genome replication
• viral production

Acknowledgments

We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Disclosure statement

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

Author contributions

J.H.Y. designed the experiments and wrote the article. J.H.Y. and S.C.H. analyzed the results. J.H.Y, W.B.H, X.L.M, and T.Z. conducted the experiments. W.Y.Z., Z.Y.W., F.M.S., S.X.Z., Y.S., Z.L.L., X.Y.Y., and X.F.Y. prepared the viruses, cell lines, and reagents, and discussed the data.

Data availability statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. The data used to support the findings of this study are available at

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Supplementary material

Supplemental data for this article can be accessed here

Additional information

Funding

This work was supported by the National Natural Science Foundation of China under Grant [numbers Major Research Plan 81871634 and Young Scientists Fund 81301416 to J.H.Y.]; the Postdoctoral Science Foundation of China under Grant [numbers Postdoctoral Research Foundation of China 2014M561302 and 2015T80299] to J.H.Y.; the Norman Bethune Program of Jilin University [Grant number 2015202] to J.H.Y.; the Jilin Provincial Science and Technology Department under Grant [numbers 20140204004YY, 20160414025GH, 20190304064YY, and 20200801020GH] to J.H.Y.; and the Department of Human Resources and Social Security of Jilin Province under Grant [number 2016014] to J.H.Y. In this study, there is no bias associated with funding. The funding institutions did not influence the study design, analysis of data, or interpretation of the results.