Bovine viral diarrhoea virus (BVDV) does not use macropinocytosis or claveolae-mediated endocytic pathway for entry into bovine or ovine cells while brefeldin A inhibits its release in ovine cells

POOJA DUBEY; NIRANJAN MISHRA; S P BEHERA; ANIL PRAKASH

doi:10.56093/ijans.v85i6.49276

Authors

POOJA DUBEY Ph.D. Scholar, ICAR-National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh 462 022 India
NIRANJAN MISHRA Principal Scientist, ICAR-National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh 462 022 India
S P BEHERA SRF, ICAR-National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh 462 022 India
ANIL PRAKASH Professor, Department of Microbiology, Barkatullah University, Bhopal

https://doi.org/10.56093/ijans.v85i6.49276

Keywords:

BVDV, Caveolae-mediated endocytosis, Entry, Macropinocytosis, Release

Abstract

Present knowledge on bovine viral diarrhoea virus (BVDV) entry, morphogenesis and release in host cells is incomplete. This study reports the results of effect of drug cytochalasin D and nystatin on BVDV entry in bovine and ovine cells and effect of Brefeldin-A (BFA), on BVDV release in ovine cells. The bovine (MDBK) and ovine (SFT-R) cells were treated with various concentrations of cytochalasin D or nystatin before infection with BVDV and at 16 h post infection, the number of infected cells was determined by immunochemistry. The ovine cells were infected with BVDV before addition of different concentration of BFA at 8 h post infection and the supernatants at 24 hpi were subjected to RT-PCR or immunochemistry. The results showed that only a minor inhibitory effect was observed on the entry and infectivity of BVDV in both ovine and bovine cells even at highest concentration of cytochalasin D or nystatin. As revealed by RT-PCR and virus titration, BFA treatment inhibited the BVDV secretion in a dose-dependent manner with complete inhibition obtained by 2μg/ml of BFA. However, the intracellular virus particles in BFA treated cell pellet were infectious. Taken together, our results showed that entry of BVDV into bovine or ovine cells is not dependent on macropinocytosis or claveolae-mediated endocytosis and provided evidence of existence of a common mechanism of BVDV release in bovine and ovine cells.

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References

Aleksandrowicz P, Marzi A, Biedenkopf N, Beimforde N, Becker S, Hoenen T, Feldmann H and Schnittler H J. 2011. Ebola virus enters host cells by macropinocytosis and clathrinmediated endocytosis. Journal of Infectious Diseases 204: S957–67. DOI: https://doi.org/10.1093/infdis/jir326

Arnal M, Fernandez-de-Luco D, Riba L, Maley M, Gilray J, Willoughby K, Vilcek S and Nettleton P F. 2004. A novel pestivirus associated with deaths in Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Journal of General Virology 85: 3653–57. DOI: https://doi.org/10.1099/vir.0.80235-0

Dasgupta A and Wilson D W. 2001. Evaluation of the primary effect of brefeldin A treatment upon herpex simplex virus assembly. Journal of General Virology 82: 1561–67. DOI: https://doi.org/10.1099/0022-1317-82-7-1561

Flanagan M D and Lin S. 1980. Cytochalasins block actin filament elongation by binding to high-affinity sites associated with Factin. Journal of Biological Chemistry 255: 835–38. DOI: https://doi.org/10.1016/S0021-9258(19)86105-7

Gray E W and Nettleton P F. 1987. The ultrastructure of cell cultures infected with border disease and bovine virus diarrhoea viruses. Journal of General Virology 68: 2339–46. DOI: https://doi.org/10.1099/0022-1317-68-9-2339

Grummer B, Beer M, Liebler-Tenorio E and Greiser-Wilke I. 2001. Localisation of viral proteins in cells infected with bovine viral diarrhoea virus. Journal of General Virology 82: 2597–605. DOI: https://doi.org/10.1099/0022-1317-82-11-2597

Gu B, Mason P, Wang L, Norton P, Bourne N, Moriarty R, Mehta A, Despande M, Shah M and Block T. 2006. Antiviral profiles of novel iminocyclitol compounds against bovine viral diarrhea virus, west Nile virus, dengue virus and hepatitis B virus. Antiviral Chemistry and Chemotherapy 18: 49–59. DOI: https://doi.org/10.1177/095632020701800105

King A M Q, Adams M J, Carstens E B and Lefkowitz E J (Eds). 2011. Virus Taxonomy: Classification and Nomenclature of Viruses. Ninth Report of the International Committee on Taxonomy of Viruses. Academic Press, San Diego, p 1003– 20.

Krey T, Thiel H J and Rumenapf T. 2005. Acid-resistant bovine pestivirus requires activation for pH-triggered fusion during entry. Journal of Virology 79: 4191–200. DOI: https://doi.org/10.1128/JVI.79.7.4191-4200.2005

Lad V J and Gupta A K. 2011. Effect of brefeldin A and monensin on Japanese encephalitis virus maturation and virus release from cells. Microbiology Research 2e9: 33–36. DOI: https://doi.org/10.4081/mr.2011.e9

Liang D, Sainz I F, Ansari I H, Gil L H V G, Vassilev V and Donis R O. 2003. The envelope glycoprotein E2 is a determinant of cell culture tropism in ruminant pestiviruses. Journal of General Virology 84: 1269–74. DOI: https://doi.org/10.1099/vir.0.18557-0

Mackenzie J M, Jones M K and Westaway E G. 1999. Markers for trans-Golgi membranes and the intermediate compartment localize to induced membranes with distinct replication functions in flavivirus-infected cells. Journal of Virology 73: 9555–67. DOI: https://doi.org/10.1128/JVI.73.11.9555-9567.1999

Macovei A, Zitzmann N, Lazar C, Dwek R A and Branza-Nichita N. 2006. Brefeldin A inhibits pestivirus release from infected cells, without affecting its assembly and infectivity. Biochemical and Biophysical Ressearch Communications 346: 1083–90. DOI: https://doi.org/10.1016/j.bbrc.2006.06.023

Mathapati B S, Mishra N, Rajukumar K, Nema R K, Behera S P and Dubey S C. 2009. Entry of bovine viral diarrhea virus into ovine cells occurs through clathrin-dependent endocytosis and low pH-dependent fusion. In Vitro Cell Development Biology Animal 46: 403–07. DOI: https://doi.org/10.1007/s11626-009-9263-9

Mercer J and Helenius A. 2009. Virus entry by macropinocytosis. Nature Cell Biology 11: 510 –20. DOI: https://doi.org/10.1038/ncb0509-510

Mercer J, Schelhaas M and Helenius A. 2010. Virus entry by endocytosis. Annual Review of Biochemistry 79: 803–33. DOI: https://doi.org/10.1146/annurev-biochem-060208-104626

Meyers G and Thiel H J. 1996. Molecular characterization of pestivirus. Advances in Virus Research 47: 53–68. DOI: https://doi.org/10.1016/S0065-3527(08)60734-4

Mishra N, Pattnaik B, Vilcek S, Patil S S, Jain P, Swamy N, Bhatia S and Pradhan H K. 2004. Genetic typing of bovine viral diarrhoea virus isolates from India. Veterinary Microbiology 104: 207–12. DOI: https://doi.org/10.1016/j.vetmic.2004.08.003

Mishra N, Rajukumar K, Vilcek S, Tiwari A, Satav J S and Dubey S C. 2008. Molecular characterization of bovine viral diarrhoea virus type 2 isolate originating from a native Indian sheep (Ovis aries). Veterinary Microbiology 130: 88–98. DOI: https://doi.org/10.1016/j.vetmic.2008.01.005

Mishra N, Pitale S S, Rajukumar K, Prakash A, Behera S P, Nema R K and Dubey S C. 2012. Genetic variety of bovine viral diarrhoea virus 1 strains isolated from sheep and goats in India. Acta Virologica 56: 209–15. DOI: https://doi.org/10.4149/av_2012_03_209

Nuttall PA. 1980. Growth characteristics of two strains of bovine virus diarrhea virus. Archives of Virology 66: 365–69. DOI: https://doi.org/10.1007/BF01320633

Pelham H R. 1991. Recycling of proteins between the endoplasmic reticulum and Golgi complex. Current Opinion in Cell Biology 3: 585–91. DOI: https://doi.org/10.1016/0955-0674(91)90027-V

Pelkmans L and Helenius A. 2003. Insider information: what viruses tell us about endocytosis. Current Opinion in Cell Biology 15: 414–22. DOI: https://doi.org/10.1016/S0955-0674(03)00081-4

Schnitzer J E, Oh P, Pinney E and Allard J. 1994. Filipin-sensitive caveolae-mediated transport in endothelium reduced transcytosis, scavenger endocytosis, and capillary permeability of select macromolecules. Journal of Cell Biology 127: 1217– 32. DOI: https://doi.org/10.1083/jcb.127.5.1217

Sieczkarski S B and Whittaker G R. 2002. Influenza virus can enter and infect cells in the absence of clathrin-mediated endocytosis. Journal of Virology 76: 10455–64. DOI: https://doi.org/10.1128/JVI.76.20.10455-10464.2002

Suksanpaisan L, Susantad T and Smith D R. 2009. Characterization of dengue virus entry into HepG2 cells. Journal of Biomedical Science 16: 17–23. DOI: https://doi.org/10.1186/1423-0127-16-17

Wang H, Yang P, Liu K, Guo F, Zhang Y, Zhang G and Jiang C. 2008. SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway. Cell Research 18: 290–301. DOI: https://doi.org/10.1038/cr.2008.15