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Journal of Virology, March 2003, p. 3882-3887, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3882-3887.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Role of the Virus Nucleoprotein in the Regulation of Lymphocytic Choriomeningitis Virus Transcription and RNA Replication

Daniel D. Pinschewer,{dagger} Mar Perez, and Juan Carlos de la Torre*

Department of Neuropharmacology, The Scripps Research Institute, La Jolla, California 92037

Received 19 September 2002/ Accepted 20 November 2002

The prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) has a bisegmented negative-strand RNA genome. Each segment carries two viral genes in opposite orientation and separated by an intergenic region (IGR). The RNA-dependent RNA polymerase (RdRp) L of LCMV produces subgenomic mRNA and full-length genomic and antigenomic RNA species in two different processes termed transcription and replication, respectively. It is widely accepted that intracellular nucleoprotein (NP) levels regulate these two processes. Intracellular NP levels increase during the course of the infection, resulting in the unfolding of secondary RNA structures within the IGR. Structure-dependent transcription termination at the IGR is thereby attenuated, promoting replication of genome and antigenome RNA species. To test this hypothesis, we established a helper-virus-free minigenome (MG) system where intracellular synthesis of an S segment analogue from a plasmid is driven by RNA polymerase I. Cotransfection with two additional plasmids expressing the minimal viral trans-acting factors L and NP under control of RNA polymerase II allowed for RNA synthesis mediated by the intracellularly reconstituted LCMV polymerase. Both processes, transcription and replication, were strictly dependent on NP. However, both were equally enhanced by incrementally increasing amounts of NP up to levels in the range of those in LCMV-infected cells. Our data are consistent with a central role for NP in transcription and replication of the LCMV genome, but they do not support the participation of NP levels in balancing the two processes.

* Corresponding author. Mailing address: Department of Neuropharmacology, IMM6, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-2152. Fax: (858) 784-9981. E-mail: juanct{at}scripps.edu.

{dagger} Present address: Department of Pathology, Institute of Experimental Immunology, University Hospital of Zurich, 8091 Zürich, Switzerland.

Journal of Virology, March 2003, p. 3882-3887, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3882-3887.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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