MICROSCOPY OF INTERNAL STRUCTURES OF SENDAI VIRUS ASSOCIATED WITH THE CYTOPLASMIC SURFACE OF HOST MEMBRANES
MICROSCOPY OF INTERNAL STRUCTURES OF SENDAI VIRUS ASSOCIATED WITH THE CYTOPLASMIC SURFACE OF HOST MEMBRANES
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DOI：
10.1016/0042-6822(82)90036-8
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发表时间：
1982-01-01
期刊：
影响因子：
3.7
通讯作者：
BACHI, T
中科院分区：
文献类型：
--
作者：
BUECHI, M;BACHI, T
用免疫荧光显微镜和冷冻干燥电镜研究了仙台病毒感染细胞质膜的胞质表面。在细胞已经附着到玻璃盖玻片上之后，使它们经受生理缓冲液的射流，其剪切掉每个细胞的上部，留下附着的膜与暴露的PS。这种脱帽操作允许直接检查与宿主细胞内表面相关的内部病毒特异性元件。在病毒出芽发生的感染阶段，观察到核蛋白（RNP）链附着在质膜的PS上。RNP粘附到膜上的位点被以正交图案排列的病毒特异性颗粒修饰。冷冻断裂膜的疏水结构域中存在相同的晶体结构，表明它们被插入到内部脂质小叶中。表面糖蛋白刺突和内部RNP与该晶体结构的空间关联表明其与内部病毒基质（M）蛋白的特殊关系（如果不是同一性的话）。这种结构元件的定位和结晶性质相对于病毒形态发生，溶血和细胞融合活动的可能意义进行了讨论。与感染细胞中观察到的上述变化相反，在与外源病毒体融合的正常细胞的PS上未发现可检测到的病毒抗原。在这些情况下，PS中不存在内化抗原可能表明潜在宿主细胞以不同于人红细胞中观察到的方式处理感染性病毒组分。在后一种情况下，病毒与细胞融合后的内化抗原仍与PS结合。
The cytoplasmic surface (PS) of the plasma membrane of cells infected with Sendai virus was studied by immunofluorescence microscopy and freeze-drying electron microscopy. After cells had been attached to glass coverslips, they were subjected to a jet stream of physiological buffer which sheared off the upper portion of each cell, leaving the attached membrane with the PS exposed. This uncapping maneuver permitted direct examination of internal virus-specific elements associated with the inner surface of the host cell. At a stage of infection at which viral budding occurs, strands of nucleoprotein (RNP) were observed to be attached to the PS of plasma membranes. The sites at which RNP was adherent to the membrane were modified by virus-specific particles arranged in orthogonal patterns. The presence of the same crystalline structures in the hydrophobic domain of freeze-fractured membranes indicated that they were inserted into the inner lipid leaflet. The spatial association of the surface glycoprotein spikes and the internal RNP with this crystalline structure suggests its special relationship if not identity with the internal viral matrix (M) protein. The possible significance of the localization and crystalline nature of this structural element with respect to viral morphogenesis, hemolytic and cell-fusing activities is discussed. In contrast to the foregoing changes observed in the infected cell, no detectable viral antigens were found on the PS of normal cells to which exogenous virions had been fused. Absence of internalized antigen from the PS under these circumstances could indicate that infectious viral components are processed by the potential host cell in a manner which differs from what is observed with human erythrocytes. In the latter instance internalized antigens after fusion of virus to the cell remain associated with the PS.