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首页> 外文期刊>Journal of Molecular Biology >The 12 A structure of trypsin-treated measles virus N-RNA.
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The 12 A structure of trypsin-treated measles virus N-RNA.

机译：胰蛋白酶处理的麻疹病毒AND-RNA的12 A结构。

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摘要

Recombinant measles virus nucleoprotein (N) was produced in insect cells where it bound to cellular RNA to form helical N-RNA structures. These structures were observed by electron microscopy but were too flexible for high-resolution image analysis. Removal of the C-terminal tail of N by trypsin treatment resulted in structures that were much more rigid and seemed more regular. Several methods of image analysis were employed in order to make a helical reconstruction of the digested N-RNA. During this analysis, it became clear that the apparently regular coils of digested N-RNA consisted of a series of closely related helical states. The iterative helical real space reconstruction method allowed the identification of two helical states for which a reconstruction could be calculated. The model with the highest resolution shows N monomers that consist of three domains and that are connected to their neighbours by two narrow connections, one close to the helical axis and another toward the middle of the monomers. There are no connections between N molecules in subsequent helical turns. After labelling the RNA in the structure with cis-platinum, the connection closest to the helical axis increased in density, suggesting the position of the RNA. The shapes of the monomers of the nucleoproteins of influenza virus, rabies virus (both determined before) and that of measles virus (determined here) are all similar, whereas the overall shapes of their respective N-RNAs (nucleocapsids) is very different. This is probably due to the position and number of the connections between the N subunits in the N-RNA, one for influenza virus allowing much flexibility, two for rabies virus at either end of the N molecules leading to ribbons and two for measles virus leading to the typical paramyxovirus helical nucleocapsid.
机译：重组麻疹病毒核蛋白（N）在昆虫细胞中产生，并与细胞RNA结合形成螺旋N-RNA结构。这些结构通过电子显微镜观察到，但是对于高分辨率图像分析来说太灵活了。用胰蛋白酶处理去除N的C末端尾巴会导致结构更加坚硬，并且看起来更规则。为了对消化的N-RNA进行螺旋重建，采用了几种图像分析方法。在此分析过程中，很明显，已消化的N-RNA的明显规则的卷曲由一系列紧密相关的螺旋状态组成。螺旋实际空间的迭代重建方法允许识别两个螺旋状态，可以针对它们计算重建。分辨率最高的模型显示N个单体由三个域组成，并通过两个狭窄的连接点与其邻居连接，一个靠近螺旋轴，另一个靠近中间。在随后的螺旋匝中，N个分子之间没有连接。用顺铂标记结构中的RNA后，最接近螺旋轴的连接的密度增加，表明RNA的位置。流感病毒，狂犬病病毒（之前已确定）和麻疹病毒（此处确定）的核蛋白单体的形状都相似，而它们各自的N-RNA（核衣壳）的总体形状却非常不同。这可能是由于N-RNA中N个亚基之间的连接位置和数量，一个是流感病毒，具有很大的灵活性，两个是在狂犬病毒的N分子两端产生带状，而两个是在麻疹病毒的引导下到典型的副粘病毒螺旋核衣壳。

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