The 3' end of Turnip crinkle virus contains a highly interactive structure including a translational enhancer that is disrupted by binding to the RNA-dependent RNA polymerase.
Precise temporal control is needed for RNA viral genomes to translate sufficient replication-required products before clearing ribosomes and initiating replication. A 3' translational enhancer in Turnip crinkle virus (TCV) overlaps an internal T-shaped structure (TSS) that binds to 60S ribosomal subunits. The higher-order structure in the region was examined through alteration of critical sequences revealing novel interactions between an H-type pseudoknot and upstream residues, and between the TSS and internal and terminal loops of an upstream hairpin. Our results suggest that the TSS forms a stable scaffold that allows for simultaneous interactions with external sequences through base pairings on both sides of its large internal symmetrical loop. Binding of TCV RNA-dependent RNA polymerase (RdRp) to the region potentiates a widespread conformational shift with substantial rearrangement of the TSS region, including the element required for efficient ribosome binding. Degrading the RdRp caused the RNA to resume its original conformation, suggesting that the initial conformation is thermodynamically favored. These results suggest that the 3' end of TCV folds into a compact, highly interactive structure allowing RdRp access to multiple elements including the 3' end, which causes structural changes that potentiate the shift between translation and replication.
Yuan
Xuefeng
X
Department of Cell Biology and Molecular Genetics, University of Maryland College Park, College Park, Maryland 20742, USA.
Shi
Kerong
K
Meskauskas
Arturas
A
Simon
Anne E
AE
D001483:Base Sequence; D019184:Carmovirus / Q000737:chemistry* / Q000235:genetics; D004742:Enhancer Elements, Genetic; D009154:Mutation; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000737:chemistry; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D014158:Transcription, Genetic
D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase