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Specific elimination of coxsackievirus B3 infected cells with a protein engineered toxin-antitoxin system

Molecular & Cellular Toxicology volume 15, pages 425–430 (2019)Cite this article

Abstract

Backgrounds

Coxsackievirus B3 (CVB3) is a member of the family Picornaviridae, and along with polio-viruses, belongs to the Enterovirus genus. The CVB3 genome is composed single-stranded RNA encoding polyproteins, which are cleaved to individual functional proteins by 2A and 3C proteases proteins which have been targeted for drug development. Here, we showed that protease activity required to activate a toxic protein may be used to prevent viral infection.

Methods

We modified the MazE-MazF antitoxin-toxin system of Escherichia coli to fuse a C-terminal fragment of MazE to the N-terminal end of toxin MazF with a linker having a specific protease cleavage site for CVB3. This fusion protein formed a stable dimer and was capable of inactivating the mRNA interferase activity of MazF which cleaves the ACA sequence in mRNA substrates.

Results

The incubation of 2A proteases with the fusion proteins induced cleavage between the MazE and MazF fragments from the fusion proteins; the subsequent release of MazF significantly inhibited virus replication. Additionally, we note that, CVB3 infected HeLa cells quickly died through a MazF toxin mediated effect before virus protein expression.

Conclusion

These findings suggest that the MazEF fusion protein has a strong potential to be developed as an anti-virus therapy following CVB3 infection.

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Acknowledgements

Jung-Ho Park and Jin-Ho Park contributed equally to this study. This study was supported by grants from the National Research Foundation (NRF) of Korea provided by the Korean Government (No. NRF-2016R1D1A1A02937046, Lim BK), the Next-Generation BioGreen 21 Program (Project No. PJ01368601), Rural Development Administration, Republic of Korea, and the KRIBB Research Initiative Program.

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Author notes

  1. These authors contributed equally to this work.

Affiliations

  1. Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, 28120, Republic of Korea

    Jung-Ho Park & Wonho Choi

  2. Department of Biomedical Science, Jungwon University, Goesan-gun, Chungbuk, 28024, Republic of Korea

    Jin-Ho Park & Byung-Kwan Lim

Authors
  1. Jung-Ho Park
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  2. Jin-Ho Park
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  3. Wonho Choi
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  4. Byung-Kwan Lim
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Corresponding author

Correspondence to Byung-Kwan Lim.

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Conflict of Interest

Jung-Ho Park, Jin-Ho Park, Wonho Choi, & Byung-Kwan Lim declares that they have no conflict of interest.

Human and animal rights

The article does not contain any studies with human and animal and this study was performed following institutional and national guidelines.

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Park, JH., Park, JH., Choi, W. et al. Specific elimination of coxsackievirus B3 infected cells with a protein engineered toxin-antitoxin system. Mol. Cell. Toxicol. 15, 425–430 (2019). https://doi.org/10.1007/s13273-019-0046-4

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  • DOI: https://doi.org/10.1007/s13273-019-0046-4

Keywords

  • Coxsackievirus
  • MazEF
  • Antitoxin
  • Cardiotropic
  • Polyprotein
  • Proteases