VP1, the Putative RNA-Dependent RNA Polymerase of Infectious Bursal Disease Virus, Forms Complexes with the Capsid Protein VP3, Leading to Efficient Encapsidation into Virus-Like Particles

Author:

Lombardo Eleuterio1,Maraver Antonio1,Castón José R.2,Rivera José1,Fernández-Arias Armando3,Serrano Antonio4,Carrascosa José L.2,Rodriguez José F.1

Affiliation:

1. Departments of Biologı́a Molecular y Celular,1

2. Estructura de Macromoléculas,2 and

3. Centro Nacional de Sanidad Agropecuaria, Apdo 10, San José de las lajas, La Habana, Cuba3

4. Inmunologı́a y Oncologı́a,4 Centro Nacional de Biotecnologı́a, Cantoblanco, 28049 Madrid, Spain, and

Abstract

ABSTRACT A cDNA corresponding to the coding region of VP1, the putative RNA-dependent RNA polymerase, of infectious bursal disease virus (IBDV) was cloned and inserted into the genome of a vaccinia virus inducible expression vector. The molecular mass and antigenic reactivity of VP1 expressed in mammalian cells are identical to those of its counterpart expressed in IBDV-infected cells. The results presented here demonstrate that VP1 is efficiently incorporated into IBDV virus-like particles (VLPs) produced in mammalian cells coexpressing the IBDV polyprotein and VP1. Incorporation of VP1 into VLPs requires neither the presence of IBDV RNAs nor that of the nonstructural polypeptide VP5. Immunofluorescence, confocal laser scanning microscopy, and immunoprecipitation analyses conclusively showed that VP1 forms complexes with the structural polypeptide VP3. Formation of VP1-VP3 complexes is likely to be a key step for the morphogenesis of IBDV particles.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

Reference44 articles.

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3. Castón J. R. and J. F. Rodriguez. Unpublished results.

4. Dobos P. Berthiaume L. Leong J. A. Kibenge K. S. B. Muller H. Nicholson B. L. Family Birnaviridae Virus taxonomy. Sixth report of the International Committee on Taxonomy of Viruses. Murphy F. A. Fauquet C. M. Bishop D. H. L. Ghabrial S. A. Jarvis A. W. Martelli G. P. Mayo M. A. Summers M. D. 1995 240 244 Springer-Verlag New York N.Y

5. Early steps in reovirus infection are associated with dramatic changes in supramolecular structure and protein conformation: analysis of virions and subviral particles by cryoelectron microscopy and image reconstruction;Dryden K. A.;J. Cell Biol.,1993

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