Kingston R L, Olson N H, Vogt V M
Institute of Molecular Biology, Howard Hughes Medical Institute, University of Oregon, Eugene, OR 97403, USA.
J Struct Biol. 2001 Oct;136(1):67-80. doi: 10.1006/jsbi.2001.4423.
We have studied the organization of mature infectious Rous sarcoma virus (RSV), suspended in vitreous ice, using transmission electron microscopy. The enveloped virions are spherical in shape, have a mean diameter of 127 nm, and vary significantly in size. Image processing reveals the presence of the viral matrix protein underlying the lipid bilayer and the viral envelope proteins external to the lipid bilayer. In the interior of the virus, the characteristic mature retroviral core is clearly imaged. In contrast to lentiviruses, such as human immunodeficiency virus, the core of RSV is essentially isometric. The capsid, or external shell of the core, has a faceted, almost polygonal appearance in electron micrographs, but many capsids also exhibit continuous surface curvature. Cores are not uniform in size or shape. Serrations observed along the projected faces of the core suggest a repetitive molecular structure. Some isolated cores were observed in the sample, confirming that cores are at least transiently stable in the absence of the viral envelope. Using an approach grounded in geometric probability, we estimate the size of the viral core from the projection data. We show that the size of the core is not tightly controlled and that core size and virion size are positively correlated. From estimates of RNA packing density we conclude that either the RNA within the core is loosely packed or, more probably, that it does not fill the core.
我们利用透射电子显微镜研究了悬浮于玻璃态冰中的成熟传染性劳氏肉瘤病毒(RSV)的结构。包膜病毒粒子呈球形,平均直径为127纳米,大小差异显著。图像处理显示脂质双层下方存在病毒基质蛋白,脂质双层外部存在病毒包膜蛋白。在病毒内部,特征性的成熟逆转录病毒核心清晰成像。与慢病毒(如人类免疫缺陷病毒)不同,RSV的核心基本呈等轴状。核心的衣壳或外壳在电子显微照片中呈现出多面的、几乎多边形的外观,但许多衣壳也呈现出连续的表面曲率。核心的大小和形状并不均匀。在核心的投影面上观察到的锯齿状表明存在重复的分子结构。在样品中观察到一些分离的核心,证实了核心在没有病毒包膜的情况下至少是暂时稳定的。我们采用基于几何概率的方法,从投影数据估算病毒核心的大小。我们发现核心大小不受严格控制,且核心大小与病毒粒子大小呈正相关。根据RNA堆积密度的估算,我们得出结论,要么核心内的RNA堆积松散,要么更有可能的是,RNA并未填满核心。