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Amsacta moorei entomopoxvirus encodes a protein kinase with dual activity and a broad substrate spectrum including two putative cellular substrates

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Abstract

Amsacta moorei entomopoxvirus (AMEV) is a poxvirus that can only infect insects. This virus is an attractive research material because it is similar to smallpox virus. AMEV is one of many viruses that encode protein kinases that drive the host's cellular mechanisms, modifying immune responses to it, and regulating viral protein activity. We report here the functional characterization of a serine/threonine (Ser/Thr) protein kinase (PK) gene (ORF AMV197) of AMEV. Expression of the AMV197 gene in baculovirus expression system yielded a ~ 35.5 kDa protein. PK activity of expressed AMV197 was shown by standard PK assay. Substrate profiling of AMV197 protein by peptide microarray indicated that the expressed protein phosphorylated 81 of 624 substrates which belong to 28 families of PK substrates. While the hypothetical AMV197 protein phosphorylates Ser/Thr only, we demonstrated that the expressed PK also phosphorylates probes with tyrosine residues on the array which is a rare property among PKs. Pull-down assay of the AMV197 protein with the subcellular protein fractionations of Ld652 cells showed that it is using two cellular proteins (18 and 42 kDa) as novel putative substrates. Our results suggest that AMEV can regulate cellular mechanisms by phosphorylating cellular proteins through AMV197 PK. However, further experiments are needed to identify the exact role of this PK in the replication of AMEV.

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Funding

This study has been funded by The Scientific and Technological Research Council of Türkiye (Project No: 110T887).

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Authors and Affiliations

  1. Faculty of Sciences, Department of Molecular Biology and Genetic, Karadeniz Technical University, 61080, Trabzon, Türkiye

    Hacer Muratoğlu

  2. Faculty of Sciences, Department of Biology, Karadeniz Technical University, 61080, Trabzon, Türkiye

    Remziye Nalcacioglu & Zihni Demirbag

  3. Laboratory for Molecular Virology, Great Lakes Forestry Centre, Sault Ste. Marie, ON, Canada

    Basil M. Arif

Authors
  1. Hacer Muratoğlu
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  2. Remziye Nalcacioglu
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  3. Basil M. Arif
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  4. Zihni Demirbag
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Contributions

Resources: ZD Conceived and designed the experiments: HM Performed the experiments: HM Analysed the data: HM, RN, BA Wrote the paper: HM, ZD, BA Review and editing: HM, ZD

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Correspondence to Zihni Demirbag.

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Edited by Sassan Asgari .

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Muratoğlu, H., Nalcacioglu, R., Arif, B.M. et al. Amsacta moorei entomopoxvirus encodes a protein kinase with dual activity and a broad substrate spectrum including two putative cellular substrates. Virus Genes 60, 287–294 (2024). https://doi.org/10.1007/s11262-024-02069-4

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  • DOI: https://doi.org/10.1007/s11262-024-02069-4

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