Cryo-Electron Microscopy of the Translational Apparatus: Experimental Evidence for the Paths of mRNA, tRNA, and the Polypeptide Chain

Cryo-Electron Microscopy of the Translational Apparatus: Experimental Evidence for the Paths of mRNA, tRNA, and the Polypeptide Chain
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翻译装置的冷冻电子显微镜:mRNA、tRNA 和多肽链路径的实验证据

DOI:
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发表时间:
2000
期刊:
影响因子:
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通讯作者:
R. Agrawal
R. Agrawal
中科院分区:
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文献类型:
--
作者:
J. Frank;R. Grassucci;A. Heagle;C. Spahn;P. Penczek;R. Agrawal

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蛋白质合成是一个将两个相当长的线性结构(mRNA和新生多肽链)和大的蛋白质因子(EF-G和氨酰-tRNA链EF-Tu GTP三元复合物)紧密结合在一起的过程,蛋白质合成提出了一个交通控制的逻辑问题:如何保证不间断的高精度性能,而没有空间干扰和各种配体的纠缠。由于冷冻电子显微镜(cryo-EM)可视化提供了第一个详细的三维(3-D)图像的核糖体,大量的工作已经进入映射的tRNA和延伸因子结合到核糖体在延长周期的各个阶段。最近的一项研究发现,70 S核糖体携带一个基因插入的tRNA样RNA片段,提供了一个更高分辨率(17-A)的空核糖体图谱,在减法中使用这个新图谱产生了一个线性质量分布,覆盖了mRNA路径的平台侧片段,以及一个质量徘徊在30 S亚基通道的入口处。结合到核糖体的tRNA已经通过3-D cryo-EM在各种tRNA-核糖体复合物中直接可视化。核糖体结合的tRNA的可视化仍然是一项具有挑战性的任务,因为分子的最小尺寸是cryo-EM分辨率的量级,并且一些tRNA结合状态的占用率本质上是低的。
As a process that brings together, in close proximity, two linear structures of considerable length (mRNA and the nascent polypeptide chain) and large protein factors (EF-G and aminoacyl-tRNA·EF-Tu·GTP ternary complex), protein synthesis poses a logistic problem of traffic control: how to guarantee uninterrupted, high-precision performance without steric interference and entanglement of the various ligands. Since cryo-electron microscopy (cryo-EM) visualization provided the first detailed three-dimensional (3-D) images of the ribosome, much work has gone into the mapping of tRNA and elongation factors bound to the ribosome at various stages of the elongation cycle. A recent study of a 70S ribosome carrying a genetically inserted tRNA-like RNA fragment furnished a higher-resolution (17-A) map of the vacant ribosome, and the use of this new map in the subtraction produced a linear mass distribution covering the platform side segment of the mRNA path, as well as a mass hovering just at the entrance of the 30S subunit channel. tRNA bound to the ribosome has been directly visualized by 3-D cryo-EM in various tRNA-ribosome complexes. Visualization of the ribosome-bound tRNA is still a challenging task because the smallest dimension of the molecule is on the order of the resolution of cryo-EM and the occupancy of some tRNA binding states is intrinsically low.
核糖体的三维冷冻电子显微镜。
DOI: 10.1016/s0076-6879(00)17020-x
发表时间: 2000
影响因子: --
作者:
Frank,J;Penczek,P;Agrawal,RK;Grassucci,RA;Heagle,AB
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功能普遍性和进化多样性:来自核糖体结构的见解。
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发表时间: 1998
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影响因子: --
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通过基因标记和冷冻电子显微镜对核糖体内的 RNA 螺旋进行直接三维定位和阳性鉴定。
DOI: 10.1016/s0969-2126(00)88347-1
发表时间: 1999
期刊: Structure (London, England : 1993)
影响因子: --
作者:
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通讯作者: Frank,J
DOI: 10.1126/science.278.5346.2123
发表时间: 1997-12-19
期刊: SCIENCE
影响因子: 56.9
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DOI: 10.1073/pnas.95.11.6134
发表时间: 1998-05-26
影响因子: 11.1
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通讯作者: Frank, J