Development and evaluation of a MAb based competitive-ELISA using helicase domain of NS3 protein for sero-diagnosis of bovine viral diarrhea in cattle and buffaloes

2008 ◽  
Vol 85 (1) ◽  
pp. 39-45 ◽  
Author(s):  
S. Bhatia ◽  
Richa Sood ◽  
N. Mishra ◽  
B. Pattnaik ◽  
H.K. Pradhan
Keyword(s):  
Competitive Elisa ◽  
Bovine Viral Diarrhea ◽  
Helicase Domain ◽  
Viral Diarrhea ◽  
Ns3 Protein

Mapping of two antigenic domains on the NS3 protein of the pestivirus bovine viral diarrhea virus

2005 ◽  
Vol 108 (1-2) ◽  
pp. 13-22 ◽  
Author(s):  
Dirk Deregt ◽  
Edward J. Dubovi ◽  
Michael E. Jolley ◽  
Phuong Nguyen ◽  
Kimberley M. Burton ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Ns3 Protein ◽  
Antigenic Domains ◽  
Viral Diarrhea Virus

scholarly journals Assignment of the Multifunctional NS3 Protein of Bovine Viral Diarrhea Virus during RNA Replication: an In Vivo and In Vitro Study

1999 ◽  
Vol 73 (11) ◽  
pp. 9196-9205 ◽  
Author(s):  
Claus W. Grassmann ◽  
Olaf Isken ◽  
Sven-Erik Behrens
Keyword(s):  
Rna Replication ◽  
Rna Helicase ◽  
Bovine Viral Diarrhea ◽  
Helicase Activity ◽  
Terminal Part ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Ns3 Protein ◽  
Viral Diarrhea Virus

ABSTRACT Studies on the replication of the pestivirus bovine viral diarrhea virus (BVDV) were considerably facilitated by the recent discovery of an autonomous subgenomic BVDV RNA replicon (DI9c). DI9c comprises mainly the untranslated regions of the viral genome and the coding region of the nonstructural proteins NS3, NS4A, NS4B, NS5A, and NS5B. To assess the significance of the NS3-associated nucleoside triphosphatase/helicase activity during RNA replication and to explore other functional features of NS3, we generated a repertoire of DI9c derivatives bearing in-frame mutations in different parts of the NS3 coding unit. Most alterations resulted in deficient replicons, several of which encoded an NS3 protein with an inhibited protease function. Three lesions permitted replication, though at a lower level than that of the wild-type RNA, i.e., replacement of the third position of the DEYH helicase motif II by either T or F and an insertion of four amino acid residues in the C-terminal part of NS3. While polyprotein proteolysis was found to be almost unaffected in these latter replicons, in vitro studies with the purified mutant NS3 proteins revealed a significantly impaired helicase activity for the motif II substitutions. NS3 with a DEFH motif, moreover, showed a significantly lower ATPase activity. In contrast, the C-terminal insertion had no negative impact on the ATPase/RNA helicase activity of NS3. All three mutations affected the synthesis of both replication products—negative-strand intermediate and progeny positive-strand RNA—in a symmetric manner. Unexpectedly, various attempts to rescue or enhance the replication capability of nonfunctional or less functional DI9c NS3 derivatives, respectively, by providing intact NS3 intrans failed. Our experimental data thus demonstrate that the diverse enzymatic activities of the NS3 protein—in particular the ATPase/RNA helicase—play a pivotal role even during early steps of the viral replication pathway. They may further indicate the C-terminal part of NS3 to be an important functional determinant of the RNA replication process.

Mapping B-cell linear epitopes of NS3 protein of bovine viral diarrhea virus

2013 ◽  
Vol 151 (3-4) ◽  
pp. 331-336 ◽  
Author(s):  
Yan Li ◽  
Ying Jia ◽  
Kai Wen ◽  
Hua Liu ◽  
Mingchun Gao ◽  
...  
Keyword(s):  
B Cell ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Ns3 Protein ◽  
Linear Epitopes ◽  
Viral Diarrhea Virus

scholarly journals The RNA Helicase and Nucleotide Triphosphatase Activities of the Bovine Viral Diarrhea Virus NS3 Protein Are Essential for Viral Replication

2000 ◽  
Vol 74 (4) ◽  
pp. 1794-1800 ◽  
Author(s):  
Baohua Gu ◽  
Changbao Liu ◽  
Juili Lin-Goerke ◽  
Derrick R. Maley ◽  
Lester L. Gutshall ◽  
...  
Keyword(s):  
Point Mutations ◽  
Rna Helicase ◽  
Bovine Viral Diarrhea ◽  
Helicase Activity ◽  
Minus Strand ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Ns3 Protein ◽  
Viral Diarrhea Virus

ABSTRACT Helicase/nucleoside triphosphatase (NTPase) motifs have been identified in many RNA virus genomes. Similarly, all the members of theFlaviviridae family contain conserved helicase/NTPase motifs in their homologous NS3 proteins. Although this suggests that this activity plays a critical role in the viral life cycle, the precise role of the helicase/NTPase in virus replication or whether it is essential for virus replication is still unknown. To determine the role of the NS3 helicase/NTPase in the viral life cycle, deletion and point mutations in the helicase/NTPase motifs of the bovine viral diarrhea virus (BVDV) (NADL strain) NS3 protein designed to abolish either helicase activity alone (motif II, DEYH to DEYA) or both NTPase and helicase activity (motif I, GKT to GAT and deletion of motif VI) were generated. The C-terminal domain of NS3 (BVDV amino acids 1854 to 2362) of these mutants and wild type was expressed in bacteria, purified, and assayed for RNA helicase and ATPase activity. These mutations behaved as predicted with respect to RNA helicase and NTPase activities in vitro. When engineered back into an infectious cDNA for BVDV (NADL strain), point mutations in either the GKT or DEYH motif or deletion of motif VI yielded RNA transcripts that no longer produced infectious virus upon transfection of EBTr cells. Further analysis indicated that these mutants did not synthesize minus-strand RNA. These findings represent the first report unequivocably demonstrating that helicase activity is essential for minus-strand synthesis.

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