VPg, coat protein and five non-structural proteins of potato A potyvirus bind RNA in a sequence-unspecific manner

The potato A potyvirus (PVA)-encoded proteins P1, HC-Pro, P3, Cl, VPg, NlaP ro, Nlb and coat protein (CP) were expressed as 6 x His-tagged recombinant proteins in Escherichia coli and purified to homogeneity. RNA binding was t ested using purified proteins in Northwestern and liquid assays. PVA protei ns except P3 bound to positive- and negative-sense transcripts prepared fro m the nontranslated 5'- and 3'-regions of PVA genomic RNA and to full-lengt h transcripts of PVA RNA. RNA binding by these proteins showed no sequence specificity since they also bound to various non-PVA control RNAs. Binding properties of P1, HC-Pro, Cl and NlaPro are consistent with previous studie s carried out on a few other potyviruses, but the binding of VPg, Nlb and C P to RNA reveal novel interactions between RNA and potyvirus proteins. Furt hermore, the RNA-binding properties of all major proteins of a poty-virus h ave not been reported previously.