Torrisi M R, Bonatti S
J Cell Biol. 1985 Oct;101(4):1300-6. doi: 10.1083/jcb.101.4.1300.
Sindbis virus-infected baby hamster kidney cells were analyzed by thin section fracture-label. Specific immunolabel with antiviral glycoprotein antibodies or with conventional lectin label (wheat germ agglutinin) were used in conjunction with colloidal gold-conjugated protein A or ovomucoid, respectively. In addition, intact infected cells were analyzed with both labeling procedures. Experiments with Sindbis infected-chick embryo fibroblast cells were carried out as controls. Viral transmembrane glycoproteins appeared present in freeze-fractured inner and outer nuclear membrane, endoplasmic reticulum, Golgi stacks and vesicles, and plasma membranes; a clear preferential partition with the exoplasmic faces of all intracellular membranes was observed. By contrast, at the plasma membrane level, Sindbis glycoproteins were found to partition preferentially with the protoplasmic face. It seems likely that this protoplasmic partition is related to the binding with the nucleocapsid that takes place during the budding of the virus. At the cell surface, viral glycoproteins always appeared clustered and were predominantly associated with budding figures: moreover, large portions of the plasma membrane were devoid of both glycoproteins and budding viruses.
通过超薄切片断裂标记法对辛德毕斯病毒感染的幼仓鼠肾细胞进行分析。分别使用抗病毒糖蛋白抗体的特异性免疫标记或传统凝集素标记（麦胚凝集素），并结合胶体金偶联的蛋白A或卵类粘蛋白。此外，对完整的感染细胞进行了两种标记程序的分析。以感染辛德毕斯病毒的鸡胚成纤维细胞进行实验作为对照。病毒跨膜糖蛋白出现在冷冻断裂的内外核膜、内质网、高尔基体堆叠和囊泡以及质膜中；观察到在所有细胞内膜的外质面上有明显的优先分布。相比之下，在质膜水平，发现辛德毕斯糖蛋白优先与原生质面分布。这种原生质分布似乎与病毒出芽过程中与核衣壳的结合有关。在细胞表面，病毒糖蛋白总是呈簇状出现，并且主要与出芽形态相关；此外，大部分质膜既没有糖蛋白也没有出芽病毒。
