Proteomic and phosphoproteomic analyses reveal several events involved in the early stages of bovine herpesvirus 1 infection

  • Marcos J. Magalhães-Junior
  • Maria Cristina Baracat-PereiraEmail author
  • Lorena K. J. Pereira
  • Camilo E. Vital
  • Marcus R. Santos
  • Pricila S. Cunha
  • Kenner M. Fernandes
  • Gustavo C. Bressan
  • Juliana L. R. Fietto
  • Abelardo Silva-Júnior
  • Márcia R. Almeida
Original Article


Herpesviruses are predicted to express more than 80 proteins during their infection cycle. The proteins synthesized by the immediate early genes and early genes target signaling pathways in host cells that are essential for the successful initiation of a productive infection and for latency. In this study, proteomic and phosphoproteomic tools showed the occurrence of changes in Madin-Darby bovine kidney cells at the early stage of the infection by bovine herpesvirus 1 (BoHV-1). Proteins that had already been described in the early stage of infection for other herpesviruses but not for BoHV-1 were found. For example, stathmin phosphorylation at the initial stage of infection is described for the first time. In addition, two proteins that had not been described yet in the early stages of herpesvirus infections in general were ribonuclease/angiogenin inhibitor and Rab GDP dissociation inhibitor beta. The biological processes involved in these cellular responses were repair and replication of DNA, splicing, microtubule dynamics, and inflammatory responses. These results reveal pathways that might be used as targets for designing antiviral molecules against BoHV-1 infection.


  • BoHV-1 infection at early stages influenced various biological processes.

  • BoHV-1 infection at early stages showed proteins reported for other virus and stages.

  • Proteins not yet reported for BoHV-1 infection in the early stage were ribonuclease/angiogenin inhibitor and Rab GDP dissociation inhibitor beta



two-dimensional gel electrophoresis






alpha-2-HS glycoprotein


apolipoprotein A-I


bovine herpesvirus 1


bovine respiratory disease complex


heterogeneous nuclear ribonucleoprotein C


3-3-cholamidopropyl dimethylammonio-1-propanesulfonate


cyprinid herpesvirus 2




deoxyuridine triphosphate


deoxyuridine triphosphatase


Epstein-Barr virus nuclear antigen 2


early gene


formic acid


glyceraldehyde-3-phosphate dehydrogenase


78 kDa glucose-regulated protein


human herpesvirus 1


heterogeneous nuclear ribonucleoprotein K


hours postinfection


heat shock protein beta-1


heat shock protein 70


HHV-1 entry mediator


immediate-early gene


immobilized pH gradient


late gene


Madin-Darby bovine kidney


minimum essential medium


multiplicity of infection


mass spectrometry






osteoclast stimulating factor


proliferating cell nuclear antigen


purine nucleoside phosphorylase


prostaglandin E synthase 3


real-time quantitative PCR


ribonuclease/angiogenin inhibitor 1


ribosomal protein large P0


ribosomal protein S18




stathmin 1


tubulin beta


trifluoroacetic acid


virion host shut-off


viral ribonucleoproteins



The authors thank the Brazilian Agencies Foundation for Research Support of Minas Gerais (FAPEMIG: Fellowships and Grants, PPM-00796-15), the Financier of Studies and Projects (FINEP: CT-INFRA/UFV-2004/2007/2008), the National Council for Scientific and Technological Development (CNPq: Grants: 483976/2012-1 and 455318/2014-0), and Coordination for the Improvement of Higher Education Personnel (CAPES: Fellowships) for financial support. The authors would like to thank Nucleus of Analysis of Biomolecules (NuBioMol, UFV, Viçosa-MG, Brazil) for assistance with mass spectrometric analysis, and the Institute of Biotechnology Applied to Agriculture (BIOAGRO, UFV, Viçosa-MG, Brazil) for technical support. Funding Sources in Brazil: Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Financiadora de Estudos e Projetos (FINEP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Author contributions

MJMJ, LKJP, CEV, and KMF cultured and infected cells, performed electrophoresis assays and protein analysis. MJMJ, PSC and MRS performed real-time PCR. MCBP, ASJ, GCB, JLRF, PSC, MRS, and MRA designed and conducted the biological assays. MJMJ, MCBP, ASJ, PSC, and MRS wrote and revised the manuscript. All authors contributed and gave approval to the final version of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors have nothing to disclose as conflicts of interest. The authors declare no competing financial interest.

Supplementary material

705_2019_4452_MOESM1_ESM.docx (3.2 mb)
Supplementary material 1 (DOCX 3289 kb)


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Copyright information

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

Authors and Affiliations

  • Marcos J. Magalhães-Junior
    • 1
    • 2
  • Maria Cristina Baracat-Pereira
    • 2
    • 7
    Email author
  • Lorena K. J. Pereira
    • 2
  • Camilo E. Vital
    • 3
  • Marcus R. Santos
    • 4
  • Pricila S. Cunha
    • 6
  • Kenner M. Fernandes
    • 5
  • Gustavo C. Bressan
    • 1
  • Juliana L. R. Fietto
    • 1
  • Abelardo Silva-Júnior
    • 4
  • Márcia R. Almeida
    • 1
  1. 1.Laboratory of Animal Molecular InfectologyFederal University of ViçosaViçosaBrazil
  2. 2.Laboratory of Proteomics and Protein BiochemistryFederal University of ViçosaViçosaBrazil
  3. 3.Nucleus of Biomolecules AnalysisFederal University of ViçosaViçosaBrazil
  4. 4.Laboratory of Immunobiology and Animal VirologyFederal University of ViçosaViçosaBrazil
  5. 5.Laboratory of Cell BiologyFederal University of ViçosaViçosaBrazil
  6. 6.Laboratory of Cell and Molecular ImmunologyFederal University of Minas GeraisBelo HorizonteBrazil
  7. 7.Department of Biochemistry and Molecular BiologyFederal University of ViçosaViçosaBrazil

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