A Replicon Trans-Packaging System Reveals the Requirement of Nonstructural Proteins for the Assembly of Bovine Viral Diarrhea Virus (BVDV) Virion

Authors

• Delin Liang, Ceres, Inc. 1535 Rancho Conejo Blvd, Thousand Oaks, CA 91320, USA
• Limei Chen, Influenza Division, NCIRD, CCID, Centers for Disease Control and Prevention, 1600 Clifton Road - Mail Stop G-16, Atlanta, GA 30333, USA
• Israrul H. Ansari, University of Nebraska - LincolnFollow
• Laura H. V. G. Gil, Oswaldo Cruz Foundation (FIOCRUZ), Aggeu Magalhães Center, Av. Professor Moraes Rego, s/n CEP- 50670-420; Recife-PE, Brazil
• Christina L. Topliff, University of Nebraska - LincolnFollow
• Clayton L. Kelling, University of Nebraska - LincolnFollow
• Ruben O. Donis, Influenza Division, NCIRD, CCID, Centers for Disease Control and PreventionFollow

Document Type

Article

Date of this Version

5-10-2009

Comments

Published in Virology 387:2 (May 10, 2009), pp. 331–340; doi 10.1016/j.virol.2009.02.019 Copyright © 2009 Elsevier Inc. Used by permission.

Abstract

A selective trans-packaging system was developed to produce and isolate bovine viral diarrhea virus (BVDV) pseudo-particles with complementing reporter replicons and their packaging proteins expressed in trans with recombinant vaccinia virus. The encapsidation of replicon rNS3-5B was dependent not only on the in trans expression of structural proteins C, Erns, E1 and E2, but also the nonstructural proteins, p7 and contiguous precursor NS2-3-4A. Nonstructural p7, NS4B, NS5A or NS5B could be expressed in cis and in trans with precursor NS2-3-4A without significantly affecting virion assembly efficiency. NS2-3-4A was identified as an in trans functional precursor in virion assembly. BVDV genomes with mutant NS5B, which did not undergo active replication, were packaged 5-fold less efficiently than the intact genomes demonstrating the importance of replication in virion packaging. These results suggest that genome replication and assembly are closely associated, consistent with a model in which these two steps are coupled for maximum efficiency.