Differential use of 3'CITEs by the subgenomic RNA of Pea enation mosaic virus 2

The genomic RNA (gRNA) of Pea enation mosaic virus 2 (PEMV2) is the template for p33 and −1 frameshift product p94. The PEMV2 subgenomic RNA (sgRNA) encodes two overlapping ORFs, p26 and p27, which are required for movement and stability of the gRNA. Efficient translation of p33 requires two of three 3' proximal cap-independent translation enhancers (3'CITEs): the kl-TSS, which binds ribosomes and engages in a long-distance interaction with the 5'end; and the adjacent eIF4E-binding PTE. Unlike the gRNA, all three 3'CITEs were required for efficient translation of the sgRNA, which included the ribosome-binding 3'TSS. A hairpin in the 5' proximal coding region of p26/p27 supported translation by the 3'CITEs by engaging in a long-distance RNA:RNA interaction with the kl-TSS. These results strongly suggest that the 5' ends of PEMV2 gRNA and sgRNA connect with the 3'UTR through similar long-distance interactions while having different requirements for 3'CITEs.