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支持脚手架的RNA的高分辨率冷EM结构确定

Daniel B Haack¹, Boris Rudolfs¹, Shouhong Jin²

¹Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA.

bioRxiv : the preprint server for biology

|June 25, 2024

概括

这项研究提出了一种新的冷EM策略,用于高分辨率地可视化小RNA,例如胺酸盐 (TPP) рибо开关. 这种方法克服了以前的局限性,使RNA分子的详细结构分析成为可能.

科学领域:

结构生物学结构生物学
在RNA生物学,RNA生物学.
生物化学生物化学

背景情况:

确定无蛋白RNA,特别是小RNA的高分辨率冷电子显微镜 (cryo-EM) 结构是一项挑战.
现有的方法通常产生低至中等分辨率,较小的宏分子缺乏核酸水平的细节.

研究的目的:

开发一种新的高分辨率冷EM结构确定小RNA的新策略.
为了证明这种方法的实用性,使用氨酸酸盐 (TPP) рибо开关.

主要方法:

采用了一种涉及将小RNA与II组内核融合的策略.
用冷电磁波测定了TPP核糖转换器在结合体和无结合体状态中的结构.

主要成果:

确定了TPP核子开关的高分辨率 (2.5 Å) 低温电磁结构,包括其连接体结合口袋.
结构揭示了在结合器结合时的aptamer域中的大规模构造变化.
无带的阿波状态结构也得到了阐明.

结论:

开发的融合策略使小RNA的前所未有的高分辨率冷EM可视化成为可能.
这种方法为确定 рибо开关结构设定了新的标准,并且适用于其他以前难以处理的小RNA.
RNA-小分子结合可以诱导影响基因表达的显著构造变化.

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相关概念视频

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...