GSE98956
RNA sequencing of plant microRNAs precipitated by the tombusviral suppressor of RNA silencing p19.
Viral suppressors of RNA silencing, VSRs, counteract the antiviral RNA silencing of host plants by sequestration of virus-derived siRNAs. A central question concerns whether and how VSRs associate cellular miRNAs and thus modulate plant gene expression during a viral infection. In this study we characterize the binding behaviour of the tombusviral p19 protein to miRNAs by performing an RNA-pull down experiment with bead-associated p19 protein from carnation italian ringspot virus. For this, we used cytoplasmatic extracs of Nicotiana tabacum protoplasts as an RNA source. By applying Next Generation Sequencing (NGS) to the precipitated small RNAs, we were able to identify miRNAs specifically associating with the protein and other that were not efficiently bound by p19. Overall design: Small RNAs were precipitated from Nicotiana tabacum cytoplasmatic BY2 lysate by carnation italian ringspot virus p19 protein. NGS-sequenced small RNAs were compared to the input RNA.