Archives of Virology

, Volume 164, Issue 1, pp 255–260 | Cite as

Bovine viral diarrhea virus non-structural protein NS4B induces autophagosomes in bovine kidney cells

  • Yuto Suda
  • Shin Murakami
  • Taisuke HorimotoEmail author
Brief Report


Bovine viral diarrhea virus (BVDV) is an important pathogen in cattle that causes economic losses in livestock industries. Autophagy is an essential cell system for the maintenance of homeostasis and is induced by various triggers, including infection by viruses. BVDV infection leads to autophagy in order to enhance its replication in cells. In this study, we investigated the effect of BVDV non-structural proteins on the induction of autophagosomes. We found that NS4B alone could induce autophagosomes, suggesting a novel and important function of NS4B in BVDV replication.



We thank Dr. Atsushi Miyawaki (Riken, Japan) for the Venus/pCS2 plasmid.

Compliance with ethical standards

Conflict of interest

There is no conflict of interest associated with this article.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent is not required because no human participants were involved in this article.


  1. 1.
    Baker JC (1995) The clinical manifestation of bovine viral diarrhea infection. Vet Clin N Am Food Anim Pract 11:425–445CrossRefGoogle Scholar
  2. 2.
    Brodersen BW (2014) Bovine viral diarrhea virus infections: manifestations of infection and recent advances in understanding pathogenesis and control. Vet Pathol 51:453–464CrossRefGoogle Scholar
  3. 3.
    Behrens SE, Grassmann CW, Thiel HJ, Meyers G, Tautz N (1998) Characterization of an autonomous subgenomic pestivirus RNA replicon. J Virol 72:2364–2372Google Scholar
  4. 4.
    Tomassini JE, Boots E, Gan L, Graham P, Munshi V, Wolanski B, Fay JF, Getty K, LaFemina R (2003) An in vitro Flaviviridae replicase system capable of authentic RNA replication. Virology 313:274–285CrossRefGoogle Scholar
  5. 5.
    Wiskerchen M, Collett MS (1991) Pestivirus gene expression: protein p80 of bovine viral diarrhea virus is a proteinase involved in polyprotein processing. Virology 184:341–350CrossRefGoogle Scholar
  6. 6.
    Warrener P, Collett MS (1995) Pestivirus NS3 (p80) protein possesses RNA helicase activity. J Virol 69:1720–1726Google Scholar
  7. 7.
    Tamura JK, Warrener P, Collett MS (1993) RNA-stimulated NTPase activity associated with the p80 protein of the pestivirus bovine viral diarrhea virus. Virology 193:1–10CrossRefGoogle Scholar
  8. 8.
    Yamane D, Zahoor MA, Mohamed YM, Azab W, Kato K, Tohya Y, Akashi H (2009) Inhibition of sphingosine kinase by bovine viral diarrhea virus NS3 is crucial for efficient viral replication and cytopathogenesis. J Biol Chem 284:13648–13659CrossRefGoogle Scholar
  9. 9.
    Tautz N, Kaiser A, Thiel HJ (2000) NS3 serine protease of bovine viral diarrhea virus: characterization of active site residues, NS4A cofactor domain, and protease–cofactor interactions. Virology 273:351–363CrossRefGoogle Scholar
  10. 10.
    Mohamed YM, Bangphoomi N, Yamane D, Suda Y, Kato K, Horimoto T, Akashi H (2014) Physical interaction between bovine viral diarrhea virus nonstructural protein 4A and adenosine deaminase acting on RNA (ADAR). Arch Virol 159:1735–1741CrossRefGoogle Scholar
  11. 11.
    Weiskircher E, Aligo J, Ning G, Konan KV (2009) Bovine viral diarrhea virus NS4B protein is an integral membrane protein associated with Golgi markers and rearranged host membranes. Virol J 6:185CrossRefGoogle Scholar
  12. 12.
    Qu L, McMullan LK, Rice CM (2001) Isolation and characterization of noncytopathic pestivirus mutants reveals a role for nonstructural protein NS4B in viral cytopathogenicity. J Virol 75:10651–10662CrossRefGoogle Scholar
  13. 13.
    Isken O, Langerwisch U, Schönherr R, Lamp B, Schröder K, Duden R, Rümenapf TH, Tautz N (2014) Functional characterization of bovine viral diarrhea virus nonstructural protein 5A by reverse genetic analysis and live cell imaging. J Virol 88:82–98CrossRefGoogle Scholar
  14. 14.
    Zahoor MA, Yamane D, Mohamed YM, Suda Y, Kobayashi K, Kato K, Tohya Y, Akashi H (2010) Bovine viral diarrhea virus non-structural protein 5A interacts with NIK- and IKKbeta-binding protein. J Gen Virol 91:1939–1948CrossRefGoogle Scholar
  15. 15.
    Lai VC, Kao CC, Ferrari E, Park J, Uss AS, Wright-Minogue J, Hong Z, Lau JY (1999) Mutational analysis of bovine viral diarrhea virus RNA-dependent RNA polymerase. J Virol 73:10129–10136Google Scholar
  16. 16.
    Zhong W, Gutshall LL, Del Vecchio AM (1998) Identification and characterization of an RNA-dependent RNA polymerase activity within the nonstructural protein 5B region of bovine viral diarrhea virus. J Virol 72:9365–9369Google Scholar
  17. 17.
    He C, Klionsky DJ (2009) Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 43:67–93CrossRefGoogle Scholar
  18. 18.
    Lee YR, Lei HY, Liu MT, Wang JR, Chen SH, Jiang-Shieh YF, Lin YS, Yeh TM, Liu CC, Liu HS (2008) Autophagic machinery activated by dengue virus enhances virus replication. Virology 374:240–248CrossRefGoogle Scholar
  19. 19.
    Dreux M, Chisari FV (2009) Autophagy proteins promote hepatitis C virus replication. Autophagy 5:1224–1225CrossRefGoogle Scholar
  20. 20.
    Jin R, Zhu W, Cao S, Chen R, Jin H, Liu Y, Wang S, Wang W, Xiao G (2013) Japanese encephalitis virus activates autophagy as a viral immune evasion strategy. PLoS One 8:e52909CrossRefGoogle Scholar
  21. 21.
    Liang Q, Luo Z, Zeng J, Chen W, Foo SS, Lee SA, Ge J, Wang S, Goldman SA, Zlokovic BV, Zhao Z, Jung JU (2016) Zika virus NS4A and NS4B proteins deregulate Akt-mTOR signaling in human fetal neural stem cells to inhibit neurogenesis and induce autophagy. Cell Stem Cell 19:663–671CrossRefGoogle Scholar
  22. 22.
    Pei J, Zhao M, Ye Z, Gou H, Wang J, Yi L, Dong X, Liu W, Luo Y, Liao M, Chen J (2014) Autophagy enhances the replication of classical swine fever virus in vitro. Autophagy 10:93–110CrossRefGoogle Scholar
  23. 23.
    Su WC, Chao TC, Huang YL, Weng SC, Jeng KS, Lai MM (2011) Rab5 and Class III phosphoinositide 3-kinase Vps34 are involved in hepatitis C virus NS4B-induced autophagy. J Virol 85:10561–10571CrossRefGoogle Scholar
  24. 24.
    Wang L, Tian Y, Ou JJ (2015) HCV induces the expression of Rubicon and UVRAG to temporally regulate the maturation of autophagosomes and viral replication. PLoS Pathog 11:e1004764CrossRefGoogle Scholar
  25. 25.
    Fu Q, Shi H, Zhang H, Ren Y, Guo F, Qiao J, Jia B, Wang P, Chen C (2014) Autophagy during early stages contributes to bovine viral diarrhea virus replication in MDBK cells. J Basic Microbiol 54:1044–1052CrossRefGoogle Scholar
  26. 26.
    Fu Q, Shi H, Ren Y, Guo F, Ni W, Qiao J, Wang P, Zhang H, Chen C (2014) Bovine viral diarrhea virus infection induces autophagy in MDBK cells. J Microbiol 52:619–625CrossRefGoogle Scholar
  27. 27.
    Zhou Y, Ren Y, Cong Y, Mu Y, Yin R, Ding Z (2017) Autophagy induced by bovine viral diarrhea virus infection counteracts apoptosis and innate immune activation. Arch Virol 162:3103–3118CrossRefGoogle Scholar
  28. 28.
    Fu Q, Shi H, Shi M, Meng L, Bao H, Zhang G, Ren Y, Zhang H, Guo F, Qiao J, Jia B, Wang P, Ni W, Sheng J, Chen C (2014) Roles of bovine viral diarrhea virus envelope glycoproteins in inducing autophagy in MDBK cells. Microb Pathog 76:61–66CrossRefGoogle Scholar
  29. 29.
    Tanida I, Ueno T, Kominami E (2008) LC3 and autophagy. In: Deretic V (ed) Autophagosome and phagosome. Humana Press, New York, pp 77–88CrossRefGoogle Scholar
  30. 30.
    Nagai M, Sakoda Y, Mori M, Hayashi M, Kida H, Akashi H (2003) Insertion of cellular sequence and RNA recombination in the structural protein coding region of cytopathogenic bovine viral diarrhoea virus. J Gen Virol 84:447–452CrossRefGoogle Scholar
  31. 31.
    Rajput MKS, Abdelsalam K, Darweesh MF, Braun LJ, Kerkvliet J, Hoppe AD, Chase CCL (2017) Both cytopathic and non-cytopathic bovine viral diarrhea virus (BVDV) induced autophagy at a similar rate. Vet Immunol Immunopathol 193–194:1–9CrossRefGoogle Scholar
  32. 32.
    Maiuri MC, Zalckvar E, Kimchi A, Kroemer G (2007) Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol 8:741–752CrossRefGoogle Scholar
  33. 33.
    Djavaheri-Mergny M, Maiuri MC, Kroemer G (2010) Cross talk between apoptosis and autophagy by caspase-mediated cleavage of Beclin 1. Oncogene 29:1717–1719CrossRefGoogle Scholar
  34. 34.
    Zhou F, Yang Y, Xing D (2011) Bcl-2 and Bcl-xL play important roles in the crosstalk between autophagy and apoptosis. FEBS J 278:403–413CrossRefGoogle Scholar
  35. 35.
    Suda Y, Yamane D, Zahoor MA, Mohamed YM, Murakami S, Kato K, Akashi H, Horimoto T (2016) Unique localization of bovine viral diarrhea virus non-structural NS4B protein in infected cells. J Adv Agric 6:913–921CrossRefGoogle Scholar
  36. 36.
    Zaffagnini G, Martens S (2016) Mechanisms of selective autophagy. J Mol Biol 428:1714–1724CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Veterinary Microbiology, Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.Division of Viral Disease and Epidemiology, National Institute of Animal HealthNational Agriculture and Food Research OrganizationTsukubaJapan

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