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首页> 外文期刊>Nature >Concurrent nucleation of T6S folding and induced fit in 30S ribosome assembly
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Concurrent nucleation of T6S folding and induced fit in 30S ribosome assembly

机译：T6S折叠的同时成核和诱导装配在30S核糖体中

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摘要

Rapidly growing cells produce thousands of new ribosomes each minute, in a tightly regulated process that is essential to cell growth. How the Escherichia coli 16S ribosomal RNA and the 20 proteins that make up the 30S ribosomal subunit can assemble correctly in a few minutes remains a challenging problem, partly because of the lack of real-time data on the earliest stages of assembly. By providing snapshots of individual RNA and protein interactions as they emerge in real time, here we show that 30S assembly nucleates concurrently from different points along the rRNA. Time-resolved hydroxyl radical footprinting was used to map changes in the structure of the rRNA within 20 milliseconds after the addition of total 30S proteins. Helical junctions in each domain fold within 100 ms. In contrast, interactions surrounding the decoding site and between the 5', the central and the 3' domains require 2-200 seconds to form. Unexpectedly, nucleo-tides contacted by the same protein are protected at different rates, indicating that initial RNA-protein encounter complexes refold during assembly. Although early steps in assembly are linked to intrinsically stable rRNA structure, later steps correspond to regions of induced fit between the proteins and the rRNA.
机译：快速生长的细胞每分钟都会在对细胞生长至关重要的严格调控的过程中产生数千个新的核糖体。大肠杆菌16S核糖体RNA和组成30S核糖体亚基的20种蛋白质如何在几分钟内正确组装仍然是一个具有挑战性的问题，部分原因是缺乏最早组装阶段的实时数据。通过提供实时出现的单个RNA和蛋白质相互作用的快照，我们在这里显示30S装配体沿rRNA的不同点同时成核。在添加总的30S蛋白后的20毫秒内，使用时间分辨的羟基自由基足迹法绘制rRNA结构的变化图。每个域中的螺旋结在100毫秒内折叠。相反，围绕解码位点以及5'，中央和3'域之间的交互需要2-200秒的时间才能形成。出乎意料的是，与同一蛋白质接触的核苷酸以不同的速率受到保护，这表明初始RNA-蛋白质遇到的复合物在组装过程中会重新折叠。尽管组装的早期步骤与内在稳定的rRNA结构相关，但后续步骤对应于蛋白质与rRNA之间的诱导拟合区域。

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