This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Casabona, J. C.
Right arrow Articles by Lopez, N.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Casabona, J. C.
Right arrow Articles by Lopez, N.

 Previous Article  |  Next Article 

Journal of Virology, July 2009, p. 7029-7039, Vol. 83, No. 14
0022-538X/09/$08.00+0     doi:10.1128/JVI.00329-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The RING Domain and the L79 Residue of Z Protein Are Involved in both the Rescue of Nucleocapsids and the Incorporation of Glycoproteins into Infectious Chimeric Arenavirus-Like Particles {triangledown}

Juan Cruz Casabona,1 Jesica M. Levingston Macleod,1 Maria Eugenia Loureiro,1 Guillermo A. Gomez,2 and Nora Lopez1*

Centro de Virología Animal (CEVAN), Instituto de Ciencia y Tecnología Dr. Cesar Milstein, Consejo Nacional de Ciencia y Tecnología (CONICET), Saladillo 2468, C1440FFX Ciudad Autónoma de Buenos Aires, Argentina,1 Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC, UNC-CONICET), Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba X5000HUA, Argentina2

Received 13 February 2009/ Accepted 24 April 2009

Arenaviruses, such as Tacaribe virus (TacV) and its closely related pathogenic Junin virus (JunV), are enveloped viruses with a bipartite negative-sense RNA genome that encodes the nucleocapsid protein (N), the precursor of the envelope glycoprotein complex (GP), the polymerase (L), and a RING finger protein (Z), which is the driving force of arenavirus budding. We have established a plasmid-based system which allowed the successful packaging of TacV-like nucleocapsids along with Z and GP of JunV into infectious virus-like particles (VLPs). By coexpressing different combinations of the system components, followed by biochemical analysis of the VLPs, the requirements for the assembly of both N and GP into particles were defined. We found that coexpression of N with Z protein in the absence of minigenome and other viral proteins was sufficient to recruit N within lipid-enveloped Z-containing VLPs. In addition, whereas GP was not required for the incorporation of N, coexpression of N substantially enhanced the ratio of GP to Z into VLPs. Disruption of the RING structure or mutation of residue L79 to alanine within Z protein, although it had no effect on Z self-budding, severely impaired VLP infectivity. These mutations drastically altered intracellular Z-N interactions and the incorporation of both N and GP into VLPs. Our results support the conclusion that the interaction between Z and N is required for assembly of both the nucleocapsids and the glycoproteins into infectious arenavirus budding particles.

* Corresponding author. Mailing address: Centro de Virología Animal, Instituto de Ciencia y Tecnología Dr. Cesar Milstein, CONICET, Saladillo 2468, C1440FFX Ciudad Autónoma de Buenos Aires, Argentina. Phone and fax: 54 11 46876735. E-mail: nlopezcevan{at}centromilstein.org.ar

{triangledown} Published ahead of print on 6 May 2009.

Journal of Virology, July 2009, p. 7029-7039, Vol. 83, No. 14
0022-538X/09/$08.00+0     doi:10.1128/JVI.00329-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»