Modulation and Visualization of EF-G Power Stroke During Ribosomal Translocation.
Modulation and Visualization of EF-G Power Stroke During Ribosomal Translocation.
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核糖体易位过程中 EF-G 动力冲程的调制和可视化。
DOI：
10.1002/cbic.201900276
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Wang,Yuhong
中科院分区：
文献类型：
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作者：
Yin,Heng;Gavriliuc,Miriam;Lin,Ran;Xu,Shoujun;Wang,Yuhong
在核糖体移位过程中，延伸因子EF-G发生巨大的构象变化，同时保持与移动的tRNA的接触。我们先前测量了伴随EF‐G催化的动力冲程，这与结构研究一致。然而，动力性中风在易位保真度中的作用仍不清楚。在这里，我们报告了通过使用两种不同的技术对结构修饰的EF‐ G的功率冲程的定量测量，并揭示了功率冲程与易位效率和保真度之间的相关性。我们发现，减少功率冲程只降低易位的百分比，但不引入易位错误。已建立的EF‐G的力-结构-功能相关性表明，动力性中风驱动核糖体移位，但mRNA阅读框架可能由核糖体本身维持。此外，这里报道的显微镜检测方法可以简单地实现其他生化应用。
During ribosome translocation, the elongation factor EF‐G undergoes large conformational change while maintaining its contact with the moving tRNA. We previously measured a power stroke accompanying EF‐G catalysis, which was consistent with structural studies. However, the role of power stroke in translocation fidelity remains unclear. Here, we report quantitative measurements of the power strokes of structurally modified EF‐Gs by using two different techniques and reveal the correlation between power stroke and translocation efficiency and fidelity. We discovered that the reduced power stroke only lowered the percentage of translocation but did not introduce translocation error. The established force ‐structure–function correlation for EF‐G indicates that power stroke drives ribosomal translocation, but the mRNA reading frame is probably maintained by ribosome itself. Furthermore, the microscope detection method reported here can be simply implemented for other biochemical applications.