Identification and location of the structural glycoproteins of a tissue culture-adapted turkey enteric coronavirus.
The Minnesota strain of turkey enteric coronavirus (TCV) was grown on a human rectal tumor (HRT-18) cell line in the presence of radiolabeled amino acids and glucosamine to analyse virion structural proteins. In addition to the 52,000 unglycosylated nucleocapsid protein, three major glycoprotein species were found to be associated with the viral envelope. A predominant glycosylated protein with a molecular weight of 22-24,000 represented the transmembrane matrix protein. Larger glycoproteins with apparent molecular weights of 180-200,000 (gp 200), 120-125,000 (gp 120) and 95-100,000 (gp 100) were associated to the characteristic large bulbous projections (peplomers) located at the surface of the virion. The gp 100 and gp 120 species apparently arose from a proteolytic cleavage of gp 200, as suggested by digestion studies with trypsin and chymotrypsin. An additional large glycoprotein with mol. wt. of 140,000 (gp 140), that behaved as a disulfide-linked dimer of a 66,000 molecule, was found to be associated to granular projections located near the base of the large peplomers. Digestion studies with trypsin, bromelain and pronase demonstrated that gp 140 was related to the hemagglutinating activity of the virus. An inner membranous sac or tongue-shaped structure could be visualized in the interior of the viral particles following treatment with pronase. In contrast, trypsin or chymotrypsin treatments resulted in evaginations ("budding") on the virus surface. Progeny viral particles produced in TCV-infected cell cultures in the presence of tunicamycin lacked both types of surface projections, as demonstrated by electron microscopy and electrophoresis. The matrix protein also appeared to be reduced to its unglycosylated form, concomitant with a considerable loss of its antigenicity. Thus, with respect to its morphological and biochemical characteristics, TCV resembles viruses belonging to the group of mammalian hemagglutinating coronaviruses, but differs in that both types of envelope glycoproteins are N-glycosylated as in case of the avian infectious bronchitis virus.
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Centre de Recherche en Médecine Comparée Université du Québec, Institut Armand-Frappier, Laval-des-Rapides, Canada.
Garzon
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Tijssen
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D003332:Coronaviridae / Q000032:analysis* / Q000302:isolation & purification / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D006023:Glycoproteins / Q000032:analysis; D006031:Glycosylation; D006386:Hemagglutination Tests; D007158:Immunologic Techniques; D014764:Viral Proteins / Q000032:analysis*; D015678:Viral Structural Proteins; D014776:Virus Cultivation
D006023:Glycoproteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; D010450:Endopeptidases