Opposite effects of subverted cellular DEAD-box helicases on TBSV recombination in plants.

<p>(A) Over-expression of AtRH2 or AtRH20 was done in <i>N</i>. <i>benthamiana</i> leaves by agroinfiltration. The same leaves were agro-infiltrated to co-express CNV helper virus and the DI-AU-FP repRNA from the 35S promoter. The control samples were obtained from leaves not expressing AtRH2 or AtRH20 proteins (lanes 5–8). Total RNA was extracted from leaves 4 days after agroinfiltration. The accumulation of repRNA, recRNAs and degRNA in <i>N</i>. <i>benthamiana</i> leaves was measured by Northern blotting (top panel), see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004680#ppat.1004680.g001" target="_blank">Fig. 1</a> for details. The ribosomal RNA (rRNA) was used as a loading control and shown in agarose gel stained with ethidium-bromide (bottom panel). (B) Over-expression of AtRH20 inhibits recRNA formation from the degRNAs. AtRH20 was over-expressed in <i>N</i>. <i>benthamiana</i> leaves by agroinfiltration. The same leaves were co-agro-infiltrated to co-express CNV p33 and p92 replication proteins and the DI-RIΔ degRNA from the 35S promoter. The control samples were obtained from leaves not expressing AtRH20 protein (lanes 1–3). See further details in panel A.