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首页> 外文期刊>Molecular and Cellular Biology >Requirement of RNA Binding of Mammalian Eukaryotic Translation Initiation Factor 4GI (eIF4GI) for Efficient Interaction of eIF4E with the mRNA Cap
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Requirement of RNA Binding of Mammalian Eukaryotic Translation Initiation Factor 4GI (eIF4GI) for Efficient Interaction of eIF4E with the mRNA Cap

机译：哺乳动物真核翻译起始因子4GI（eIF4GI）RNA结合对于eIF4E与mRNA帽有效相互作用的要求

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摘要

Eukaryotic mRNAs possess a 5′-terminal cap structure (cap), m7GpppN, which facilitates ribosome binding. The cap is bound by eukaryotic translation initiation factor 4F (eIF4F), which is composed of eIF4E, eIF4G, and eIF4A. eIF4E is the cap-binding subunit, eIF4A is an RNA helicase, and eIF4G is a scaffolding protein that bridges between the mRNA and ribosome. eIF4G contains an RNA-binding domain, which was suggested to stimulate eIF4E interaction with the cap in mammals. In Saccharomyces cerevisiae, however, such an effect was not observed. Here, we used recombinant proteins to reconstitute the cap binding of the mammalian eIF4E-eIF4GI complex to investigate the importance of the RNA-binding region of eIF4GI for cap interaction with eIF4E. We demonstrate that chemical cross-linking of eIF4E to the cap structure is dramatically enhanced by eIF4GI fragments possessing RNA-binding activity. Furthermore, the fusion of RNA recognition motif 1 (RRM1) of the La autoantigen to the N terminus of eIF4GI confers enhanced association between the cap structure and eIF4E. These results demonstrate that eIF4GI serves to anchor eIF4E to the mRNA and enhance its interaction with the cap structure.
机译：真核mRNA具有5'端帽结构（cap），m 7 GpppN，有助于核糖体结合。该帽由真核翻译起始因子4F（eIF4F）绑定，该因子由eIF4E，eIF4G和eIF4A组成。 eIF4E是帽结合亚基，eIF4A是RNA解旋酶，eIF4G是在mRNA和核糖体之间架桥的支架蛋白。 eIF4G包含一个RNA结合结构域，建议在哺乳动物中刺激eIF4E与帽的相互作用。但是，在中，没有观察到这种效果。在这里，我们使用重组蛋白来重构哺乳动物eIF4E-eIF4GI复合体的帽结合，以研究eIF4GI的RNA结合区对于帽与eIF4E相互作用的重要性。我们证明，具有RNA结合活性的eIF4GI片段大大增强了eIF4E到帽结构的化学交联。此外，La自身抗原的RNA识别基序1（RRM1​​）与eIF4GI的N末端融合，可增强帽结构与eIF4E之间的结合。这些结果表明，eIF4GI可将eIF4E锚定在mRNA上并增强其与帽结构的相互作用。

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