Recruitment of the 40S ribosomal subunit by the West Nile virus 3′ UTR promotes the cross-talk between the viral genomic ends for translation regulation

Abstract

Flaviviral RNA genomes are composed of discrete RNA structural units arranged in an ordered fashion and grouped into complex folded domains that regulate essential viral functions, e.g. replication and translation. This is achieved by adjusting the overall structure of the RNA genome via the establishment of inter- and intramolecular interactions. Translation regulation is likely the main process controlling flaviviral gene expression. Although the genomic 3′ UTR is a key player in this regulation, little is known about the molecular mechanisms underlying this role. The present work provides evidence for the specific recruitment of the 40S ribosomal subunit by the 3′ UTR of the West Nile virus RNA genome, showing that the joint action of both genomic ends contributes the positioning of the 40S subunit at the 5′ end. The combination of structural mapping techniques revealed specific conformational requirements at the 3′ UTR for 40S binding, involving the highly conserved SL-III, 5′DB, 3′DB and 3′SL elements, all involved in the translation regulation. These results point to the 40S subunit as a bridge to ensure cross-talk between both genomic ends during viral translation and support a link between 40S recruitment by the 3′ UTR and translation control.