Real-Time Imaging of Translation on Single mRNA Transcripts in Live Cells.
Real-Time Imaging of Translation on Single mRNA Transcripts in Live Cells.
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DOI：
10.1016/j.cell.2016.04.040
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发表时间：
2016-05-05
期刊：
影响因子：
64.5
通讯作者：
Zhuang X
中科院分区：
文献类型：
--
作者：
Wang C;Han B;Zhou R;Zhuang X
翻译受到严格的空间和时间控制，以确保蛋白质在细胞中正确的时间和地点产生。允许实时，高分辨率可视化的活细胞中的翻译的方法是必不可少的理解时空动态的翻译调节。基于新生多肽信号的多价荧光放大，我们开发了一种方法来在活细胞中真实的时间内对单个mRNA分子的翻译进行成像，从而可以直接可视化翻译位点处的翻译事件。使用这种方法，我们监测瞬态变化的翻译动力学响应于环境应力，捕捉不同的流动性的个别多聚核糖体在不同的亚细胞室，并检测3'UTR依赖的本地翻译和主动运输的多聚核糖体在树突的初级神经元。
Translation is under tight spatial and temporal controls to ensure protein production in the right time and place in cells. Methods that allow real-time, high-resolution visualization of translation in live cells are essential for understanding the spatiotemporal dynamics of translation regulation. Based on multivalent fluorescence amplification of the nascent polypeptide signal, we develop a method to image translation on individual mRNA molecules in real time in live cells, allowing direct visualization of translation events at the translation sites. Using this approach, we monitor transient changes of translation dynamics in responses to environmental stresses, capture distinct mobilities of individual polysomes in different subcellular compartments, and detect 3’UTR-dependent local translation and active transport of polysomes in dendrites of primary neurons.