Kinetic analysis of late steps of eukaryotic translation initiation.

Kinetic analysis of late steps of eukaryotic translation initiation.
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真核翻译启动的后期步骤的动力学分析。

DOI:
10.1016/j.jmb.2008.10.029
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发表时间:
2009-01-16
影响因子:
5.6
通讯作者:
Lorsch, Jon R.
Lorsch, Jon R.
中科院分区:
生物学2区
文献类型:
--
作者:
Acker, Michael G.;Shin, Byung-Sik;Nanda, Jagpreet S.;Saini, Adesh K.;Dever, Thomas E.;Lorsch, Jon R.

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关于真核生物翻译起始晚期事件的分子机制知之甚少。我们对起始密码子识别后的预起始复合物 (PIC) 到最终 80S 起始复合物 (IC) 的转变进行了动力学剖析。由此产生的框架表明,eIF5B 实际上加速了核糖体亚基的连接速率,并且这种加速受到结合核苷酸介导的因子的 GTP 酶活性位点构象的影响。 eIF1A 通过其 C 端与 eIF5B 的相互作用加速连接,并且 eIF1A 从起始核糖体中释放(仅在亚基连接后发生)通过 eIF5B 的 GTP 水解加速。亚基连接后,eIF5B 进行的 GTP 水解改变了最终 IC 的构象,并为促进 eIF1A 的快速释放扫清了道路。我们的数据结合之前的工作表明,eIF1A 在整个启动过程中都存在于核糖体上,并在每个阶段发挥关键作用。
Little is known about the molecular mechanics of the late events of translation initiation in eukaryotes. We present a kinetic dissection of the transition from a pre-initiation complex (PIC) after start codon recognition to the final 80S initiation complex (IC). The resulting framework reveals that eIF5B actually accelerates the rate of ribosomal subunit joining and this acceleration is influenced by the conformation of the GTPase active site of the factor mediated by the bound nucleotide. eIF1A accelerates joining through its C-terminal interaction with eIF5B, and eIF1A release from the initiating ribosome, which occurs only after subunit joining, is accelerated by GTP hydrolysis by eIF5B. Following subunit joining, GTP hydrolysis by eIF5B alters the conformation of the final IC and clears a path to promote rapid release of eIF1A. Our data, coupled with previous work, indicate that eIF1A is present on the ribosome throughout the entire initiation process and plays key roles at every stage.
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