The role of the 5'-cap structure in viral ribonucleoproteins assembly from potato virus X coat protein and RNAs

Abstract

The potato virus X (PVX) virion can be reconstituted in vitro from the virus coat protein (CP) and RNA; heterologous RNAs may be used as well. In our recent study, structure and properties of cognate and heterologous viral ribonucleoproteins (vRNPs) were demonstrated to be similar to those of native virions. The assembly was found to be initiated at the 5' terminus of an RNA and was not dependent on RNA sequence. The aim of the present study was to search for a signal or an essential structural element that directs packaging of viral genetic material into vRNPs. vRNPs were formed by incubation of the PVX CP with heterologous capped RNAs, their functional fragments lacking the cap structure, as well as the capped and uncapped transcripts corresponding to the 5'-terminal region of the genomic PVX RNA. Experimental data show that the presence of the cap structure at the 5' end of a nucleic acid is an important condition for vRNP assembly from RNA and CP. Presumably, the 5'-cap affects conformational state of the RNA region responsible for the efficient interaction with CP and creates conformational encapsidation signal for vRNP assembly.

Keywords: AFM; Assembly; BMV; CP; Cap structure; NTA; PVX; Plant virus; Potato virus X; TEM; TGB; TMV; Viral ribonucleoproteins; atomic force microscopy; brome mosaic virus; coat protein; nanoparticle tracking analysis; potato virus X; tobacco mosaic virus; transmission electron microscopy; triple gene block; vRNP; viral ribonucleoprotein.