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

Selina, Polina I.; Karaseva, Maria A.; Komissarov, Alexey A.; Safina, Dina; Lunina, N. A.; Roschina, Marina P.; Demidyuk, Ilya V.; Kostrov, Sergey V.

doi:10.1038/s41598-021-97641-5

Cited by 3 publications

(2 citation statements)

References 26 publications

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“…In most positive-sense, single-stranded RNA viruses, viral 3CL pro or 3C proteases (3C pro ) cleaved the viral polyprotein at multiple conserved sites and are engaged in translational processing of the viral non-structural proteins [54] , [55] , [56] , [57] , [58] , [59] . The hydrolysis sites of 3C pro /3CL pro are broadly resembling and usually include a Gln or Glu residue at the P1 position along with a small amino acid residue downstream [34] , [60] , [61] .…”

Section: Discussionmentioning

confidence: 99%

An intermolecular salt bridge linking substrate binding and P1 substrate specificity switch of arterivirus 3C-like proteases

Chen

Zhou

Yang

et al. 2022

Computational and Structural Biotechnology Journal

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Section: Discussionmentioning

confidence: 99%

An intermolecular salt bridge linking substrate binding and P1 substrate specificity switch of arterivirus 3C-like proteases

Chen

Zhou

Yang

et al. 2022

Computational and Structural Biotechnology Journal

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“…In the diagram of abnormal animal proportions, we subtracted the percentage of defective animals in the control group from the values obtained to take the number of defective individuals among uninjected animals as the zero level. The method of embryotoxicity assessment was described in detail elsewhere [ 44 ].…”

Section: Methodsmentioning

confidence: 99%

Efficiency of Promoters of Human Genes FAP and CTGF at Organism Level in a Danio rerio Model

Selina

Alekseenko

Kurtova

et al. 2023

IJMS

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The identification of tissue-specific promoters for gene therapeutic constructs is one of the aims of complex tumor therapy. The genes encoding the fibroblast activation protein (FAP) and the connective tissue growth factor (CTGF) can function in tumor-associated stromal cells but are practically inactive in normal adult cells. Accordingly, the promoters of these genes can be used to develop vectors targeted to the tumor microenvironment. However, the efficiency of these promoters within genetic constructs remains underexplored, particularly, at the organism level. Here, we used the model of Danio rerio embryos to study the efficiency of transient expression of marker genes under the control of promoters of the FAP, CTGF, and immediate early genes of Human cytomegalovirus (CMV). Within 96 h after the injection of vectors, the CTGF and CMV promoters provided similar equal efficiency of reporter protein accumulation. In the case of the FAP promoter, a high level of reporter protein accumulation was observed only in certain zebrafish individuals that were considered developmentally abnormal. Disturbed embryogenesis was the factor of changes in the exogenous FAP promoter function. The data obtained make a significant contribution to understanding the function of the human CTGF and FAP promoters within vectors to assess their potential in gene therapy.

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Expression and Purification of His-Tagged Variants of Human Hepatitis A Virus 3C Protease

Karaseva,

Gramma,

Safina

et al. 2024

PPL

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Background: Protease 3C (3Cpro) is the only protease encoded in the human hepatitis A virus genome and is considered a potential target for antiviral drugs due to its critical role in the viral life cycle. Additionally, 3Cpro has been identified as a potent inducer of ferroptosis, a newly described type of cell death. Therefore, studying the molecular mechanism of 3Cpro functioning can provide new insights into viral-host interaction and the biological role of ferroptosis. However, such studies require a reliable technique for producing the functionally active recombinant enzyme. Objective: Here, we expressed different modified forms of 3Cpro with a hexahistidine tag on the N- or C-terminus to investigate the applicability of Immobilized Metal Ion Affinity Chromatography (IMAC) for producing 3Cpro. objective: Here, we expressed in Escherichia coli different modified forms of the 3Cpro with a hexahistidine tag on the N- or C-terminus to study the applicability of the immobilized metal ion affinity chromatography (IMAC) for 3Cpro producing. Methods: We expressed the proteins in Escherichia coli and purified them using IMAC, followed by gel permeation chromatography. The enzymatic activity of the produced proteins was assayed using a specific chromogenic substrate. method: The resulted proteins were purified using IMAC followed by gel permeation chromatography. Enzymatic activity of the produced proteins was assayed using a specific chromogenic substrate. Results: Our findings showed that the introduction and position of the hexahistidine tag did not affect the activity of the enzyme. However, the yield of the target protein was highest for the variant with seven C-terminal residues replaced by a hexahistidine sequence. Conclusion: We demonstrated the applicability of our approach for producing recombinant, enzymatically active 3Cpro.

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

Cited by 3 publications

References 26 publications

An intermolecular salt bridge linking substrate binding and P1 substrate specificity switch of arterivirus 3C-like proteases

An intermolecular salt bridge linking substrate binding and P1 substrate specificity switch of arterivirus 3C-like proteases

Efficiency of Promoters of Human Genes FAP and CTGF at Organism Level in a Danio rerio Model

Expression and Purification of His-Tagged Variants of Human Hepatitis A Virus 3C Protease

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