Zinoviev Alexandra, Hellen Christopher U T, Pestova Tatyana V
Department of Cell Biology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA.
Department of Cell Biology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA.
Mol Cell. 2015 Mar 19;57(6):1059-1073. doi: 10.1016/j.molcel.2015.01.039.
Reinitiation is a strategy used by viruses to express several cistrons from one mRNA. Although extremely weak after translation of long open reading frames (ORFs) on cellular mRNAs, reinitiation occurs efficiently on subgenomic bicistronic calicivirus mRNAs, enabling synthesis of minor capsid proteins. The process is governed by a short element upstream of the restart AUG, designated "termination upstream ribosomal binding site" (TURBS). It contains the conserved Motif 1 complementary to h26 of 18S rRNA, displayed in the loop of a hairpin formed by species-specific Motifs 2/2(∗). To determine the advantages conferred on reinitiation by TURBS, we reconstituted this process in vitro on two model bicistronic calicivirus mRNAs. We found that post-termination ribosomal tethering of mRNA by TURBS allows reinitiation by post-termination 80S ribosomes and diminishes dependence on eukaryotic initiation factor 3 (eIF3) of reinitiation by recycled 40S subunits, which can be mediated either by eIFs 2/1/1A or by Ligatin following ABCE1-dependent or -independent splitting of post-termination complexes.
重新起始是病毒用于从一个mRNA表达多个顺反子的一种策略。虽然在细胞mRNA上长开放阅读框(ORF)翻译后极其微弱,但重新起始在亚基因组双顺反子杯状病毒mRNA上高效发生,从而能够合成次要衣壳蛋白。该过程由重新起始AUG上游的一个短元件控制,该元件被称为“终止上游核糖体结合位点”(TURBS)。它包含与18S rRNA的h26互补的保守基序1,呈现在由物种特异性基序2/2(∗)形成的发夹环中。为了确定TURBS赋予重新起始的优势,我们在体外对两种双顺反子杯状病毒模型mRNA重建了这一过程。我们发现,TURBS对mRNA的终止后核糖体束缚作用允许终止后80S核糖体进行重新起始,并减少了重新起始对循环40S亚基的真核起始因子3(eIF3)的依赖性,这可以由eIFs 2/1/1A介导,或者在终止后复合物的ABCE1依赖性或非依赖性分裂后由连接蛋白介导。