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Cell-line-dependent crystal morphology and sublocalization of the Thyrinteina arnobia cypovirus polyhedrin expressed from a recombinant baculovirus

  • Leonardo A. Silva
  • Daniel M. P. Ardisson-AraújoEmail author
  • Fabricio S. Morgado
  • André B. Horta
  • Manoel Victor Franco Lemos
  • Carlos F. Wilcken
  • Bergmann M. Ribeiro
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Abstract

We describe an unexpected feature observed for the heterologous expression of the Thyrinteina arnobia cypovirus polyhedrin from a recombinant baculovirus infection in different insect cell lines. The in cellulo-formed crystals varied in size and shape depending on the cell line. Crystals formed in Trichoplusia ni-derived cells were cubic (0.1-2 μm) and localized in both the nucleus and cytoplasm, whereas those formed in Spodoptera frugiperda-derived cells were ovate and ellipsoidal (0.1-3 μm) and also localized in both the nucleus and cytoplasm. The molecular basis for differences in the morphology, size, and location of cypovirus occlusion bodies is unclear, and cellular proteins might play a role in their formation and location.

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Funding statement

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grant number 407908/2013-7) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF, grant number 193.001532/2016).

Supplementary material

705_2019_4214_MOESM1_ESM.tif (3.5 mb)
Fig. S1 (A) Transmission electron micrograph of Trichoplusia ni-derived cells (Tn5B) cells infected with the TharCPV-polh-expressing baculovirus, showing crystal formation in the cell nucleus (Nu) at 48 h postinfection. (B) Transmission electron micrograph of Spodoptera frugiperda-dervided cells (Sf21) cells infected with the TharCPV-polh-expressing baculovirus, showing crystals in both the cell cytoplasm (Cyto) and nucleus (Nu) at 48 h postinfection. The crystals are indicated by black arrowheads. (C) Immunofluorescence and bright-field micrographs of Trichoplusia ni-derived cells (Tn5B) cells infected with the TharCPV-polh-expressing baculovirus, showing crystal formation mainly in the cell nucleus at 48 h postinfection. (D) Immunofluorescence and bright-field micrographs of Spodoptera frugiperda-derived cells (Sf21 cells) infected with the TharCPV-polh-expressing baculovirus, showing crystals mostly in the cytoplasm at 48 h postinfection. Green color shows the location of ThaCPV-polyhedrin identified using an anti-his antibody and a secondary antibody conjugated with AlexaFluor 488 (TIFF 3566 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Leonardo A. Silva
    • 1
  • Daniel M. P. Ardisson-Araújo
    • 2
    Email author
  • Fabricio S. Morgado
    • 1
  • André B. Horta
    • 3
  • Manoel Victor Franco Lemos
    • 4
  • Carlos F. Wilcken
    • 3
  • Bergmann M. Ribeiro
    • 1
  1. 1.Cell Biology DepartmentUnB-University of BrasíliaBrasíliaBrazil
  2. 2.Laboratory of Insect Virology, Department of Biochemistry and Molecular BiologyFederal University of Santa MariaSanta MariaBrazil
  3. 3.Plant Protection DepartmentFCA/UNESP, São Paulo State UniversityBotucatuBrazil
  4. 4.Applied Biology DepartmentFCAV/UNESP, São Paulo State UniversityJaboticabalBrazil

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