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eIF2 interactions with initiator tRNA and eIF2B are regulated by post-translational modifications and conformational dynamics

机译：eIF2与启动子tRNA和eIF2B的相互作用受翻译后修饰和构象动力学的调节

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Translation of messenger RNA (mRNA) into proteins is key to eukaryotic gene expression and begins when initiation factor-2 (eIF2) delivers methionyl initiator tRNA (Met-tRNAi Met) to ribosomes. This first step is controlled by eIF2B mediating guanine nucleotide exchange on eIF2. We isolated eIF2 from yeast and used mass spectrometry to study the intact complex, and found that eIF2β is the most labile of the three subunits (eIF2α/β/γ). We then compared conformational dynamics of the ternary complex eIF2:GTP:Met-tRNAi Met with apo eIF2 using comparative chemical cross-linking. Results revealed high conformational dynamics for eIF2α in apo eIF2 while in the ternary complex all three subunits are constrained. Novel post-translational modifications identified here in both eIF2 and eIF2B were combined with established sites, and located within protein sequences and homology models. We found clustering at subunit interfaces and highly phosphorylated unstructured regions, at the N-terminus of eIF2β, and also between the eIF2Bε core and catalytic domains. We propose that modifications of these unstructured regions have a key role in regulating interactions between eIF2 and eIF2B, as well as other eIFs.
机译：信使RNA（mRNA）到蛋白质的翻译是真核基因表达的关键，并且始于起始因子2（eIF2）传递甲硫酰引发剂tRNA（Met-tRNA i Met ）核糖体。第一步是通过eIF2B介导eIF2上鸟嘌呤核苷酸交换来控制的。我们从酵母中分离出eIF2，并使用质谱研究了完整的复合物，发现eIF2β是三个亚基（eIF2α/β/γ）中最不稳定的。然后，我们使用比较化学交联比较了三元复合物eIF2：GTP：Met-tRNA i Met 与apo eIF2的构象动力学。结果显示，载脂蛋白eIF2中eIF2α的构象动力学很高，而在三元复合物中，所有三个亚基都受到约束。此处在eIF2和eIF2B中鉴定的新的翻译后修饰与已建立的位点结合，并位于蛋白质序列和同源性模型内。我们发现在亚基界面和高度磷酸化的非结构化区域，eIF2β的N端以及eIF2Bε核心和催化结构域之间聚集。我们建议对这些非结构化区域的修饰在调节eIF2和eIF2B以及其他eIF之间的相互作用中起关键作用。

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