The peplomers of Berne virus.
Using [3H]glucosamine and [3H]mannose labels, two virus-specific glycosylated polypeptide species with Mr values of about 200,000 (200K) and in the 75K to 100K range, respectively, were recognized in Berne virus-infected embryonic mule skin cells. In purified virions only the latter glycoprotein occurred. Concanavalin A was bound to the virion as evidenced by reduction in infectivity. Analyses using SDS-PAGE, blotting and glycoprotein identification with concanavalin A and horseradish peroxidase showed coincidence of the virion glycoprotein signals with the maximum infectivity and haemagglutinating activity in an isokinetic sucrose gradient. Polyclonal rabbit immune serum and a neutralizing and haemagglutination-inhibiting monoclonal antibody raised against Berne virus recognized both the 75K to 100K and the '200K' glycoproteins. Using tunicamycin, a concentration-dependent inhibition of infectivity was noted; however, non-infectious particles containing the two major polypeptides (20K and 22K) were released from the cells in small quantities. The glycoproteins were absent from cytoplasmic extracts and a novel polypeptide of about 150K was identified instead. Translation of poly(A)-selected intracellular RNA from infected cells in a rabbit reticulocyte cell-free system also resulted in the appearance of a new high Mr polypeptide (about 170K). Using pulse-chase labelling and radioimmunoprecipitation, suggestive evidence for a precursor-product relationship between the intracellular '200K' and the virion glycoproteins has been obtained. These experiments identify the N-glycosylated proteins in the 75K to 100K range as constituents of the peplomeric envelope projection of Berne virus; they probably arise by post-translational processing of a 150K to 170K precursor molecule involving glycosylation and subsequent cleavage.
D000914:Antibodies, Viral / Q000276:immunology; D003208:Concanavalin A / Q000494:pharmacology; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012328:RNA Viruses / Q000032:analysis* / Q000187:drug effects / Q000276:immunology; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D014779:Virus Replication / Q000187:drug effects
D000914:Antibodies, Viral; D006023:Glycoproteins; D010455:Peptides; D014759:Viral Envelope Proteins; D003208:Concanavalin A; D014415:Tunicamycin