Interactions of bovine viral diarrhoea virus glycoprotein E(rns) with cell surface glycosaminoglycans.
Recombinant E(rns) glycoprotein of bovine viral diarrhoea virus (BVDV) has been tagged with a marker epitope or linked to an immunoglobulin Fc tail and expressed in insect and mammalian cell lines. The product was shown to be functional, both having ribonuclease activity and binding to a variety of cells that were permissive and non-permissive for replication of BVDV. Addition of soluble E(rns) to the medium blocked replication of BVDV in permissive cells. Binding of epitope-tagged E(rns) to permissive calf testes (CTe) cells was abolished and virus infection was reduced when cells were treated with heparinases I or III. E(rns) failed to bind to mutant Chinese hamster ovary (CHO) cells that lacked glycosaminoglycans (pgsA-745 cells) or heparan sulphate (pgsD-677 cells) but bound to normal CHO cells. E(rns) also bound to heparin immobilized on agarose and could be eluted by heparin and by a high concentration of salt. Flow cytometric analysis of E(rns) binding to CTe cell cultures showed that glycosaminoglycans such as heparin, fucoidan and dermatan sulphate all inhibit binding but dextran sulphate, keratan sulphate, chondroitin sulphate and mannan fail to inhibit binding. The low molecular mass polysulphonated inhibitor suramin also inhibited binding to CTe cells but poly-L-lysine did not. Furthermore, suramin, the suramin analogue CPD14, fucoidan and pentosan polysulphate inhibited the infectivity of virus. It is proposed that binding of E(rns) to cells is through an interaction with glycosaminoglycans and that BVDV may bind to cells initially through this interaction.
Iqbal
M
M
Division of Molecular Biology, Institute for Animal Health, Compton Laboratory, Compton, Newbury RG20 7NN, UK.
Flick-Smith
H
H
McCauley
J W
JW
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D016466:CHO Cells; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000821:virology*; D006224:Cricetinae; D017931:DNA Primers / Q000235:genetics; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D004330:Drosophila; D005260:Female; D006025:Glycosaminoglycans / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006493:Heparin / Q000378:metabolism; D019763:Heparin Lyase / Q000494:pharmacology; D008297:Male; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D017931:DNA Primers; D006025:Glycosaminoglycans; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D006493:Heparin; D019763:Heparin Lyase