PESTIVIRUS GENE-EXPRESSION - PROTEIN P80 OF BOVINE VIRAL DIARRHEA VIRUS IS A PROTEINASE INVOLVED IN POLYPROTEIN PROCESSING

被引:147
作者
WISKERCHEN, M [1 ]
COLLETT, MS [1 ]
机构
[1] MEDIMMUNE INC, 19 FIRSTFIELD RD, GAITHERSBURG, MD 20878 USA
关键词
D O I
10.1016/0042-6822(91)90850-B
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Bovine viral diarrhea virus (BVDV), the prototypic pestivirus, possesses a positive-strand RNA genome with a single large open reading frame (ORF) encoding about 4000 amino acids. We have endeavored to elucidate the mechanisms involved in protein biogenesis by this pestivirus. Here, we present our studies on gene expression from the viral nonstructural protein coding region encompassing the car{ballot box}y-terminal 60% of the ORF. Previous sequence and modeling analyses predicted the amino-terminal region of the BVDV nonstructural protein p80 to be a trypsin-like serine proteinase. Using a mammalian cell transient expression system, we show that this region indeed possessed a proteolytic activity and, further, required the serine residue previously predicted to be the putative serine proteinase catalytic site. We found the p80-region proteinase activity was required for proteolytic processing of all viral nonstructural proteins. Cleavage by this activity at the amino and car{ballot box}y termini of the p80 protein itself likely occurred intramolecularly (in cis), since we were unable to demonstrate activityin trans at these sites. Cleavages at the three processing sites downstream of the car{ballot box}y terminus of p80 were shown to occurin trans. However, p80 proteinase activity alone was not sufficient for cleavage of the last of these sites. Another viral gene product, or specific condition, is implicated as a necessary cofactor for p80 proteinase activity at this site. Pestivirus polyprotein processing can now be compared to similar events by viruses of other groups. Finally, the potential role of the p80 proteinase activity in the phenotype of cytopathic biotypes of BVDV is discussed. © 1991 Academic Press, Inc.
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页码:341 / 350
页数:10
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