In vitro reconstitution of eukaryotic translation reveals cooperativity between release factors eRF1 and eRF3
In vitro reconstitution of eukaryotic translation reveals cooperativity between release factors eRF1 and eRF3
复制标题
DOI：
10.1016/j.cell.2006.04.035
复制
发表时间：
2006-06-16
期刊：
影响因子：
64.5
通讯作者：
Pestova, Tatyana V.
中科院分区：
文献类型：
--
作者：
Alkalaeva, Elena Z.;Pisarev, Andrey V.;Pestova, Tatyana V.
真核翻译终止是由肽释放因子ERF1和ERF3触发的。 ERF1识别所有三个终止密码子并诱导肽基TRNA的水解，而ERF3的功能仍然晦涩。在这里，我们使用纯化的核糖体亚基在体外重新构成了真核翻译的所有步骤。启动，伸长和终止因素；和氨基酸trnas。这使我们能够使用在编码四肽的mRNA上组装的早产络合物研究终止，并提出了一种翻译终止模型，该模型解释了ERF1和ERF3在确保快速释放新生多肽时的合作作用。在此模型中，ERF1，ERF3和GTP与早产配合物的结合首先诱导结构重排，该结构重排被表现为：2核苷酸向前移动，归因于早产络合物，导致GTP水解导致GTP水解，然后通过粘液中的TRNA快速水解。 ERF1和ERF3之间的合作性需要ERF1的ERF3结合c末端结构域。
Eukaryotic translation termination is triggered by peptide release factors eRF1 and eRF3. Whereas eRF1 recognizes all three termination codons and induces hydrolysis of peptidyl tRNA, eRF3's function remains obscure. Here, we reconstituted all steps of eukaryotic translation in vitro using purified ribosomal subunits; initiation, elongation, and termination factors; and aminoacyl tRNAs. This allowed us to investigate termination using pretermination complexes assembled on mRNA encoding a tetrapeptide and to propose a model for translation termination that accounts for the cooperative action of eRF1 and eRF3 in ensuring fast release of nascent polypeptide. In this model, binding of eRF1, eRF3, and GTP to pretermination complexes first induces a structural rearrangement that is manifested as a :2 nucleotide forward shift of the toeprint attributed to pretermination complexes that leads to GTP hydrolysis followed by rapid hydrolysis of pepticlyl tRNA. Cooperativity between eRF1 and eRF3 required the eRF3 binding C-terminal domain of eRF1.