Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6141522 | Virology | 2011 | 13 Pages |
Abstract
RNA-RNA recombination salvages viral RNAs and contributes to their genomic variability. A recombinationally-active subgenomic promoter (sgp) has been mapped in Brome mosaic bromovirus (BMV) RNA3 (Wierzchoslawski et al., 2004. J. Virol.78, 8552-8864) and mRNA-like 5â² sgRNA3a was characterized (Wierzchoslawski et al., 2006. J. Virol. 80, 12357-12366). In this paper we describe sgRNA3a-mediated recombination in both in vitro and in vivo experiments. BMV replicase-directed co-copying of (â) RNA3 with wt sgRNA3a generated RNA3 recombinants in vitro, but it failed to when 3â²-truncated sgRNA3a was substituted, demonstrating a role for the 3â² polyA tail. Barley protoplast co-transfections revealed that (i) wt sgRNA3a recombines at the 3â² and the internal sites; (ii) 3â²-truncated sgRNA3as recombine more upstream; and (iii) 5â²-truncated sgRNA3 recombine at a low rate. In planta co-inoculations confirmed the RNA3-sgRNA3a crossovers. In summary, the non-replicating sgRNA3a recombines with replicating RNA3, most likely via primer extension and/or internal template switching.
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Authors
Joanna Sztuba-SoliÅska, Aleksandra Dzianott, Jozef J. Bujarski,