Key result
Rotavirus NSP3 and the 3' terminal GACC sequence are essential for efficient viral mRNA translation, requiring both the eIF4G- and RNA-binding domains of NSP3.
The study demonstrates that efficient rotavirus mRNA translation requires the 3' GACC sequence, both NSP3 domains, and a good Kozak context, while tolerating very short 5' UTRs.
Hypothesis-generating for NSP3-targeted antivirals; leaves open efficacy in human rotavirus infection.
Rotavirus NSP3 is a translational surrogate of the PABP-poly(A) complex for rotavirus mRNAs. To further explore the effects of NSP3 and untranslated regions (UTRs) on rotavirus mRNAs translation, we used a quantitative in vivo assay with simultaneous cytoplasmic NSP3 expression (wild-type or deletion mutant) and electroporated rotavirus-like and standard synthetic mRNAs. This assay shows that the last four GACC nucleotides of viral mRNA are essential for efficient translation and that both the NSP3 eIF4G- and RNA-binding domains are required. We also show efficient translation of rotavirus-like mRNAs even with a 5'UTR as short as 5 nucleotides, while more than eleven nucleotides are required for the 3'UTR. Despite the weak requirement for a long 5'UTR, a good AUG environment remains a requirement for rotavirus mRNAs translation.
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Gratia et al. (2016) studied Rotavirus mRNA translation. NSP3 expression and modified untranslated regions (UTRs) vs. eGFP expression or unmodified UTRs was evaluated on Translation efficiency (measured by luciferase activity). Rotavirus NSP3 and the 3' terminal GACC sequence are essential for efficient viral mRNA translation, requiring both the eIF4G- and RNA-binding domains of NSP3.
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