BROME MOSAIC VIRUS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 我们制造了一种突变的雀麦花叶病毒(BMV)，它具有有趣的生物学特性。这些突变通过在BMV衣壳亚基的N端富含精氨酸的手臂上增加了两个α-螺旋，增加了病毒颗粒内的正电荷。BMV蛋白外壳的X射线晶体结构已经解决，但关于RNA的内层以及N末端手臂如何与RNA相互作用的信息尚不清楚。 我们提出了一项合作，我们可以使用冷冻EM来成像和重建野生型和突变型BMV。然后，我们将追踪这两个粒子的N端手臂，并确定它是如何与RNA相互作用的。 这个项目的目标是可视化这个N末端的臂，它对于与包装的病毒RNA和病毒粒子内的RNA的结构相互作用至关重要。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have made a mutant Brome mosaic virus (BMV) that has an interesting biological property. The mutations increase the positive charges inside the viral particles through the addition of two alpha-helices in the N-terminal arginine-rich arm in the capsid subunit of BMV. The X-ray crystal structure of the protein shell of BMV has been solved, but there is no information on the inner layer of RNA and how the N-terminal arm interacts with the RNA. We propose a collaboration where we can use cryo-EM to image and reconstruct the wild-type and mutant BMV. We will then trace the N-terminal arm of the two particles and identify how it interacts with RNA. The goal of this project is to visualize this N-terminal arm that is critical for interaction with the packaged viral RNAs and the structure of the RNAs inside the virion.