Gratia Matthieu, Vende Patrice, Charpilienne Annie, Baron Hilma Carolina, Laroche Cécile, Sarot Emeline, Pyronnet Stéphane, Duarte Mariela, Poncet Didier
Institut de Biologie Integrative de la Cellule (I2BC), UMR 9198, Département de Virologie, USC INRA 1358, Gif sur Yvette, France.
INSERM UMR-1037 - Université de Toulouse III-Paul Sabatier, Laboratoire d'Excellence Toulouse Cancer (TOUCAN), Equipe labellisée Ligue Contre le Cancer Toulouse, France.
PLoS One. 2016 Jan 4;11(1):e0145998. doi: 10.1371/journal.pone.0145998. eCollection 2016.
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.
轮状病毒NSP3是轮状病毒mRNA的聚腺苷酸结合蛋白（PABP）-多聚腺苷酸（poly(A)）复合物的翻译替代物。为了进一步探究NSP3和非翻译区（UTRs）对轮状病毒mRNA翻译的影响，我们采用了一种定量体内测定法，同时在细胞质中表达NSP3（野生型或缺失突变体），并对轮状病毒样mRNA和标准合成mRNA进行电穿孔。该测定表明，病毒mRNA的最后四个GACC核苷酸对于高效翻译至关重要，并且NSP3的eIF4G结合域和RNA结合域均是必需的。我们还表明，即使5'UTR短至5个核苷酸，轮状病毒样mRNA仍能高效翻译，而3'UTR则需要超过11个核苷酸。尽管对长5'UTR的需求较弱，但良好的AUG环境仍然是轮状病毒mRNA翻译的必要条件。
