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

AbstractThe 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.

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Cytotoxic effect of co-expression of human hepatitis A virus 3C protease and bifunctional suicide protein FCU1 genes in a bicistronic vector

Molecular Biology Reports ◽

10.1007/s11033-017-4113-4 ◽

2017 ◽

Vol 44 (4) ◽

pp. 323-332 ◽

Cited By ~ 1

Author(s):

Alexey Komissarov ◽

Ilya Demidyuk ◽

Dina Safina ◽

Marina Roschina ◽

Andrey Shubin ◽

...

Keyword(s):

Cytotoxic Effect ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Bicistronic Vector ◽

3C Protease ◽

Human Hepatitis

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Human hepatitis A virus 3C protease exerts a cytostatic effect on Saccharomyces cerevisiae and affects the vacuolar compartment

Biologia ◽

10.2478/s11756-020-00569-w ◽

2020 ◽

Author(s):

Andrey V. Shubin ◽

Alexey A. Komissarov ◽

Maria A. Karaseva ◽

Benjamin S. Padman ◽

Sergey V. Kostrov ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Cytostatic Effect ◽

3C Protease ◽

Vacuolar Compartment ◽

Human Hepatitis

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Restricted replication of human hepatitis A virus in cell culture: intracellular biochemical studies.

Journal of Virology ◽

10.1128/jvi.37.1.216-225.1981 ◽

1981 ◽

Vol 37 (1) ◽

pp. 216-225 ◽

Cited By ~ 22

Author(s):

S A Locarnini ◽

A G Coulepis ◽

E G Westaway ◽

I D Gust

Keyword(s):

Cell Culture ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Human Hepatitis ◽

Biochemical Studies

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Individual Expression of Hepatitis A Virus 3C Protease Induces Ferroptosis in Human Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms22157906 ◽

2021 ◽

Vol 22 (15) ◽

pp. 7906

Author(s):

Alexey A. Komissarov ◽

Maria A. Karaseva ◽

Marina P. Roschina ◽

Andrey V. Shubin ◽

Nataliya A. Lunina ◽

...

Keyword(s):

Cell Death ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Proteolytic Enzymes ◽

Morphological Changes ◽

Human Cells ◽

Mitochondrial Potential ◽

3C Protease ◽

Wide Range

Regulated cell death (RCD) is a fundamental process common to nearly all living beings and essential for the development and tissue homeostasis in animals and humans. A wide range of molecules can induce RCD, including a number of viral proteolytic enzymes. To date, numerous data indicate that picornaviral 3C proteases can induce RCD. In most reported cases, these proteases induce classical caspase-dependent apoptosis. In contrast, the human hepatitis A virus 3C protease (3Cpro) has recently been shown to cause caspase-independent cell death accompanied by previously undescribed features. Here, we expressed 3Cpro in HEK293, HeLa, and A549 human cell lines to characterize 3Cpro-induced cell death morphologically and biochemically using flow cytometry and fluorescence microscopy. We found that dead cells demonstrated necrosis-like morphological changes including permeabilization of the plasma membrane, loss of mitochondrial potential, as well as mitochondria and nuclei swelling. Additionally, we showed that 3Cpro-induced cell death was efficiently blocked by ferroptosis inhibitors and was accompanied by intense lipid peroxidation. Taken together, these results indicate that 3Cpro induces ferroptosis upon its individual expression in human cells. This is the first demonstration that a proteolytic enzyme can induce ferroptosis, the recently discovered and actively studied type of RCD.

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