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Evaluation of the anti-apoptotic activity of bovine alphaherpesvirus type 5 US3 protein kinase in insect cells using a recombinant baculovirus

  • Veterinary Microbiology - Research Paper
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Abstract

Bovine alphaherpesvirus type 5 (BoHV-5) is one of the main agents responsible for meningoencephalitis in cattle in Brazil, causing significant economic losses. It is known that other viruses of the Herpesviridae family such as Bovine alphaherpesvirus type 1, Swine alphaherpesvirus type 1, and the Human alphaherpesvirus types 1 and 2 encode genes homologous to BoHV-5, with recognized action in the control of apoptosis. The objective of this work was to express the BoHV-5 US3 gene in a baculovirus-based expression system for the production of the serine/threonine kinase protein and to evaluate its activity in the control of apoptosis in vitro. A recombinant baculovirus derived from the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) containing the US3 gene and a deletion in the baculovirus anti-apoptotic p35 gene was constructed using the Bac-to-Bac™ system. This recombinant baculovirus was used to evaluate the anti-apoptotic activity of the recombinant US3 protein in insect cells comparing with two other AcMNPV recombinants, one containing a functional copy of the AcMNPV anti-apoptotic p35 gene and an AcMNPV p35 knockout virus with the anti-apoptotic iap-3 gene from Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV). We found that the caspase level was higher in insect cells infected with the US3-contanining recombinant virus than in cells infected with the AcMNPV recombinants containing the p35 and iap-3 genes. These results indicate that the BoHV-5 US3 protein kinase gene is not able to block apoptosis in insect cells induced by the infection of a p35 knockout AcMNPV.

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Funding

This work was supported by Fundação de Apoio à Pesquisa do Distrito Federal- FAPDF (process number: 0193.001235/2016, FAPDF number: 1063/2016) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior –CAPES.

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Authors and Affiliations

  1. Laboratory of Veterinary Microbiology, Faculty of Veterinary Medicine and Agronomy, University of Brasília, Brasília, DF, Brazil

    Alice M. Silva & Simone Perecmanis

  2. Laboratory of Baculovirus, Cell Biology Department, University of Brasília, Brasília, DF, Brazil

    Fabrício S. Morgado, Leonardo A. Silva & Bergmann M. Ribeiro

  3. Veterinary Hospital, Faculty of Veterinary Medicine and Agronomy, University of Brasília, Brasília, DF, Brazil

    José R. J. Borges

  4. Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil

    Daniel M. P. Ardisson-Araújo

  5. Laboratory of Bioinformatics and Biotechnology, Federal University of Tocantins, Campus of Gurupi, Gurupi, TO, Brazil

    Fabrício S. Campos

Authors
  1. Alice M. Silva
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  2. Fabrício S. Morgado
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  3. Leonardo A. Silva
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  4. José R. J. Borges
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  5. Simone Perecmanis
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  6. Daniel M. P. Ardisson-Araújo
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  7. Bergmann M. Ribeiro
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  8. Fabrício S. Campos
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Correspondence to Fabrício S. Campos.

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Responsible Editor: Giliane Souza Trindade.

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Fig. 1

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Fig 1

qPCR quantitation of intracellular viral DNA in Sf-21 insect cells infected with different recombinant AcMNPV baculoviruses after 0, 24, 48 h.p.i. Data represent the mean (± standard deviation, SD) of independent experiments, each performed in triplicate. Error bars indicate SDs. Abbreviations: Acpg: AcPG virus; P35: AcMNPVΔP35 virus; IAP-3: AcMNPVΔP35.IAP3; US3: AcMNPVΔP35.US3. (TIF 52 kb)

Table 1

Cell lines and recombinant baculovirus (PDF 156 kb)

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Silva, A.M., Morgado, F.S., Silva, L.A. et al. Evaluation of the anti-apoptotic activity of bovine alphaherpesvirus type 5 US3 protein kinase in insect cells using a recombinant baculovirus. Braz J Microbiol 51, 827–835 (2020). https://doi.org/10.1007/s42770-019-00215-x

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  • DOI: https://doi.org/10.1007/s42770-019-00215-x

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