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Journal of Virology, December 2001, p. 12241-12251, Vol. 75, No. 24
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.24.12241-12251.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Transcription and RNA Replication of Tacaribe Virus Genome and Antigenome Analogs Require N and L Proteins: Z Protein Is an Inhibitor of These Processes

Nora López, Rodrigo Jácamo, and María T. Franze-Fernández*

Centro de Virología Animal (CONICET), Serrano 669, C1414DEM Buenos Aires, Argentina

Received 30 April 2001/Accepted 6 September 2001

Tacaribe virus (TV), the prototype of the New World group of arenaviruses, comprises a single phylogenetic lineage together with four South American pathogenic producers of hemorrhagic disease. The TV genome consists of two single-stranded RNA segments called S and L. A reconstituted transcription-replication system based on plasmid-supplied TV-like RNAs and TV proteins was established. Plasmid expression was driven by T7 RNA polymerase supplied by a recombinant vaccinia virus. Plasmids were constructed to produce TV S segment analogs containing the negative-sense copy of chloramphenicol acetyltransferase (CAT) flanked at the 5' and 3' termini by sequences corresponding to those of the 5' and 3' noncoding regions of the S genome (minigenome) or the S antigenome (miniantigenome). In cells expressing N and L proteins, input minigenome or miniantigenome produced, respectively, encapsidated miniantigenome or minigenome which in turn produced progeny minigenome or progeny miniantigenome. Both minigenome and miniantigenome in the presence of N and L mediated transcription, which was analyzed as CAT expression. Coexpression of the small RING finger Z (p11) protein was highly inhibitory to both transcription and replication mediated by the minigenome or the miniantigenome. The effect depended on synthesis of Z protein rather than on plasmid or the RNA and was not ascribed to decreased amounts of plasmid-supplied template or proteins (N or L). N and L proteins were sufficient to support full-cycle RNA replication of a plasmid-supplied S genome analog in which CAT replaced the N gene. Replication of this RNA was also inhibited by Z expression.

* Corresponding author. Mailing address: Centro de Virología Animal (CONICET), Serrano 669, C1414DEM Buenos Aires, Argentina. Phone and fax: (54)11-4856-4495 or (54)11-4825-1863. E-mail: mtfranzecevan{at}datamarkets.com.ar.

Journal of Virology, December 2001, p. 12241-12251, Vol. 75, No. 24
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.24.12241-12251.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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