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Journal of Virology, August 1999, p. 6973-6983, Vol. 73, No. 8
Departments of Biología Molecular y
Celular,1 Estructura de
Macromoléculas,2 and
Inmunología y
Oncología,4 Centro Nacional de
Biotecnología, Cantoblanco, 28049 Madrid, Spain, and
Centro Nacional de Sanidad Agropecuaria, Apdo 10, San
José de las lajas, La Habana, Cuba3
Received 19 January 1999/Accepted 11 May 1999
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.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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
*
Corresponding author. Mailing address: Departamento de
Biología Molecular y Celular, Centro Nacional de
Biotecnología, Universidad Autonoma de Madrid, Cantoblanco,
28049 Madrid, Spain. Phone: 34-91-5854558. Fax: 34-91-5854506. E-mail:
jfrodrig{at}cnb.uam.es.
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