BROME MOSAIC VIRUS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们已经取得了一个突变体雀麦花叶病毒（BMV），具有一个有趣的生物学特性。这些突变通过在BMV衣壳亚基的N-末端富含丝氨酸的臂中添加两个α-螺旋来增加病毒颗粒内的正电荷。BMV的蛋白质外壳的X射线晶体结构已经解决，但没有关于RNA内层以及N-末端臂如何与RNA相互作用的信息。 我们提出了一个合作，我们可以使用cryo-EM图像和重建野生型和突变体BMV。 然后，我们将追踪这两个粒子的N-末端臂，并确定它如何与RNA相互作用。 这个项目的目标是可视化这个N-末端臂，它对于与包装的病毒RNA的相互作用和病毒粒子内RNA的结构至关重要。

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.