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Journal of Virology, November 2004, p. 11678-11685, Vol. 78, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.21.11678-11685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Evolutionary Transition toward Defective RNAs That Are Infectious by Complementation

Juan García-Arriaza,1 Susanna C. Manrubia,2 Miguel Toja,1,{dagger} Esteban Domingo,1* and Cristina Escarmís1

Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco,1 Centro de Astrobiología (CSIC-INTA), Torrejón de Ardoz, Madrid, Spain2

Received 4 March 2004/ Accepted 14 June 2004

Passage of foot-and-mouth disease virus (FMDV) in cell culture resulted in the generation of defective RNAs that were infectious by complementation. Deletions (of nucleotides 417, 999, and 1017) mapped in the L proteinase and capsid protein-coding regions. Cell killing followed two-hit kinetics, defective genomes were encapsidated into separate viral particles, and individual viral plaques contained defective genomes with no detectable standard FMDV RNA. Infection in the absence of standard FMDV RNA was achieved by cotransfection of susceptible cells with transcripts produced in vitro from plasmids encoding the defective genomes. These results document the first step of an evolutionary transition toward genome segmentation of an unsegmented RNA virus and provide an experimental system to compare rates of RNA progeny production and resistance to enhanced mutagenesis of a segmented genome versus its unsegmented counterpart.

* Corresponding author. Mailing address: Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain. Phone: 34 91 4978485. Fax: 34 91 4974799. E-mail: edomingo{at}cbm.uam.es.

{dagger} Present address: Operon S.A., 50419-Cuarte de Huerva, Zaragoza, Spain.

Journal of Virology, November 2004, p. 11678-11685, Vol. 78, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.21.11678-11685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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