Embryotoxic activity of 3C protease of human hepatitis A virus in developing Danio rerio embryos

Polina I Selina,Alexey A Komissarov,Maria A Karaseva,Nataliya A Lunina,Eugene D Sverdlov,Ilya V Demidyuk,Sergey V Kostrov,Marina P Roschina,Dina R Safina

doi:10.1038/s41598-021-97641-5

Polina I Selina, Alexey A Komissarov + Show 7 more

Open Access

https://doi.org/10.1038/s41598-021-97641-5

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Abstract

The 3C protease is a key factor in picornavirus-induced pathologies with a comprehensive action on cell targets. However, the effects induced by the enzyme have not been described at the organismic level. Here, the model of developing Danio rerio embryos was used to analyze possible toxic effects of the 3C protease of human hepatitis A virus (3Cpro) at the whole-body level. The transient 3Cpro expression had a notable lethal effect and induced a number of specific abnormalities in Danio rerio embryos within 24 h. These effects are due to the proteolytic activity of the enzyme. At the same time, the 3Cpro variant with reduced catalytic activity (3Cmut) increased the incidence of embryonic abnormalities; however, this effect was smaller compared to the native enzyme form. While the expression of 3Cmut increased the overall rate of abnormalities, no predominance of specific ones was observed. The data obtained point to a presence significant impact of picornavirus 3Cprotease at the whole-organism level and make contribution to the study of the infectious process caused by human hepatitis A virus.

Highlights

The pathogen-host interactions are complex and realized at different levels
Using continuous in vitro cultures it has been demonstrated that the 3C protease can induce different cell death types depending on picornavirus s pecies[9,10,11,12,13,14,15,16,17,18]; its impact has not been studied at the whole-organism level
The possible embryotoxic activity of 3Cpro was evaluated by inoculation of the experimental and control constructs into the Danio rerio yolk of fertilized eggs prior to the fist division of cleavage period at 0.3 to 33 attomoles, after which the embryonic mortality and developmental abnormalities were evaluated

Summary

Introduction

The pathogen-host interactions are complex and realized at different levels. Analysis of the pathogenic mechanisms makes it possible to expose the pattern common for different agents as well as to reveal promising targets for therapeutic intervention. One should consider that the complex interaction with host cells and tissues can involve pathogen components that affect different targets The complex of such interactions defines the pathology pattern. We used a model system based on developing Danio rerio embryos to analyze possible toxic effects induced by the viral 3C protease at the whole-organism level. This protein is critical in the picornavirus-mediated pathology and seems to have an integral effect on cell targets[1,2,3,4,5,6,7,8]. We analyzed the embryotoxic effect of picornavirus 3C protease using the enzyme of human hepatitis A virus (3Cpro) inducing caspase-independent cell death in vitro[11]

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