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Journal of Virology, August 1999, p. 6973-6983, Vol. 73, No. 8
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

Eleuterio Lombardo,1 Antonio Maraver,1 José R. Castón,2 José Rivera,1 Armando Fernández-Arias,3 Antonio Serrano,4 José L. Carrascosa,2 and José F. Rodriguez1,*

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.


* 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.


Journal of Virology, August 1999, p. 6973-6983, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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Copyright © 1999 by the American Society for Microbiology. All rights reserved.