An actin-binding protein is involved in pestivirus entry into bovine cells

doi:10.1016/S0168-1702(00)00159-3

Virus Research

Volume 68, Issue 1, June 2000, Pages 1-5

https://doi.org/10.1016/S0168-1702(00)00159-3 Get rights and content

Abstract

Infection of bovine cells with bovine viral diarrhoea virus (BVDV) can be blocked by the monoclonal antibody (mab) BVD/CA 26, which is directed against a cellular membrane protein. To characterize this molecule, it was isolated and purified by column chromatography. It was found to be an acidic, glycosylated membrane protein consisting of two polypeptide chains of about 28 and 56 kDa. Under non-reducing conditions the chains formed multimers of about 200 kDa. In an actin binding assay the 56 kDa polypeptide chain bound to F-actin as judged by co-sedimentation with actin filaments. Since the target molecule of BVD/CA 26 is localized on the surface of living cells and additionally binds to F-actin, a possible biological function may be to connect the cortical actin filaments with the cellular plasma membrane. The blocking effect of BVD/CA 26 indicates that this cellular plasma membrane protein is involved in the endocytic pathway of BVDV particles.

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Acknowledgements

We thank Mrs R. Neth for excellent technical assistance and Mr M. Wolfhagen for preparation of the photographs.

References (20)

E.F. Flores et al.
Isolation of a mutant MDBK cell line resistant to bovine viral diarrhea virus infection due to a block in viral entry
Virology
(1995)
M.M. Hulst et al.
Glycoprotein E2 of classical swine fever virus: expression in insect cells and identification as a ribonuclease
Virology
(1994)
V. Moennig et al.
The pestiviruses
Adv. Virus Res.
(1992)
T.L. Poole et al.
Pestivirus translation initiation occurs by internal ribosome entry
Virology
(1995)
W. Xue et al.
Identification of bovine viral diarrhea virus receptor in different cell types
Vet. Microbiol.
(1996)
W. Xue et al.
Characterization of a putative receptor protein for bovine viral diarrhea virus
Vet. Microbiol.
(1997)
S. Bolin et al.
Monoclonal antibodies with neutralizing activity segregate isolates of bovine viral diarrhoea virus into groups
Arch. Virol.
(1988)
R.O. Donis et al.
Neutralizing monoclonal antibodies to bovine viral diarrhoea virus bind to the 56K to 58K glycoprotein
J. Gen. Virol.
(1988)
E.F. Flores et al.
Swine and ruminant pestiviruses require the same cellular factor to enter bovine cells
J. Gen. Virol.
(1996)
Francki, R.I.B., Fauquet, C.M., Knudson, D.L., Brown, F. (Eds.) (1991). Classification and nomenclature of viruses....

There are more references available in the full text version of this article.

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An actin-binding protein is involved in pestivirus entry into bovine cells

Abstract

Section snippets

Acknowledgements

Virology

Virology

Adv. Virus Res.

Virology

Vet. Microbiol.

Vet. Microbiol.

Monoclonal antibodies with neutralizing activity segregate isolates of bovine viral diarrhoea virus into groups

Arch. Virol.

Neutralizing monoclonal antibodies to bovine viral diarrhoea virus bind to the 56K to 58K glycoprotein

J. Gen. Virol.

Swine and ruminant pestiviruses require the same cellular factor to enter bovine cells

J. Gen. Virol.