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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Intersubunit movement is required for ribosomal translocation
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Intersubunit movement is required for ribosomal translocation

机译：核糖体易位需要亚基间运动

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摘要

Translocation of tRNA and mRNA during protein synthesis is believed to be coupled to structural changes in the ribosome. The "ratchet model," based on cryo-EM reconstructions of ribosome complexes, invokes relative movement of the 30S and 50S ribosomal subunits in this process; however, evidence that directly demonstrates a requirement for intersubunit movement during translocation is lacking. To address this problem, we created an intersubunit disulfide cross-link to restrict potential movement. The cross-linked ribosomes were unable to carry out polypeptide synthesis; this inhibition was completely reversed upon reduction of the disulfide bridge. In vitro assays showed that the cross-linked ribosomes were specifically blocked in elongation factor G-depen-dent translocation. These findings show that intersubunit movement is required for ribosomal translocation, accounting for the universal two-subunit architecture of ribosomes.
机译：据信在蛋白质合成过程中tRNA和mRNA的易位与核糖体的结构变化有关。基于核糖体复合物的冷冻-EM重建的“棘轮模型”在此过程中引起30S和50S核糖体亚基的相对运动。然而，缺乏直接证明易位期间亚单位间运动需求的证据。为了解决这个问题，我们创建了亚基间二硫键交联键来限制潜在的移动。交联的核糖体不能进行多肽合成。当二硫键还原时，这种抑制作用被完全逆转。体外测定显示，交联的核糖体在伸长因子G依赖性转运中被特异性阻断。这些发现表明核糖体易位需要亚基间运动，这解释了核糖体的普遍的两个亚基结构。

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