Visualizing a viral genome with contrast variation small angle X-ray scattering

Visualizing a viral genome with contrast variation small angle X-ray scattering
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通过小角度 X 射线散射对比变化可视化病毒基因组

DOI:
10.1074/jbc.ra120.013961
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发表时间:
2020
影响因子:
4.8
通讯作者:
Pollack, Lois
Pollack, Lois
中科院分区:
生物学2区
文献类型:
--
作者:
San Emeterio, Josue;Pollack, Lois

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尽管一些单链RNA病毒对人类健康构成威胁,但人们对它们的组装知之甚少。这项工作的目标是引入一种新的工具,用于观察RNA基因组通过蛋白质引导其自身的包装和组装。对比度变化小角X射线散射(CV-SAXS)是一种强大的工具,有可能通过这种组装过程来监测病毒RNA结构的变化。蛋白质虽然存在,但对测量的信号没有贡献。作为评估病毒基因组研究的可行性的第一步,用CV-SAXS专门检测了双核苷化MS 2 RNA的结构,并与来自不对称冷冻EM重建的结构进行了比较。与游离RNA的额外比较突出了衣壳蛋白诱导的显著结构重排,并邀请应用时间分辨CV-SAXS来揭示导致有效病毒组装的相互作用。
Despite the threat to human health posed by some single-stranded RNA viruses, little is understood about their assembly. The goal of this work is to introduce a new tool for watching an RNA genome direct its own packaging and encapsidation by proteins. Contrast variation small-angle X-ray scattering (CV-SAXS) is a powerful tool with the potential to monitor the changing structure of a viral RNA through this assembly process. The proteins, though present, do not contribute to the measured signal. As a first step in assessing the feasibility of viral genome studies, the structure of encapsidated MS2 RNA was exclusively detected with CV-SAXS and compared with a structure derived from asymmetric cryo-EM reconstructions. Additional comparisons with free RNA highlight the significant structural rearrangements induced by capsid proteins and invite the application of time-resolved CV-SAXS to reveal interactions that result in efficient viral assembly.
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