Assembly of enveloped viruses in Madin-Darby canine kidney cells: polarized budding from single attached cells and from clusters of cells in suspension.
Assembly of enveloped viruses in Madin-Darby canine kidney cells: polarized budding from single attached cells and from clusters of cells in suspension.
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DOI：
10.1083/jcb.96.3.866
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发表时间：
1983-03
影响因子：
7.8
通讯作者：
Sabatini, D D
中科院分区：
文献类型：
--
作者：
Rodriguez-Boulan, E;Paskiet, K T;Sabatini, D D
在狗肾上皮细胞系Madin-Darby犬肾(MDCK)融合单层中，RNA包膜病毒的组装反映了细胞的功能极化。因此，流感、仙台和猿猴病毒5从顶端(自由)表面发芽，而水疱性口炎病毒(VSV)在基底侧质膜域组装(Rodriguez-Boulan，E.和D.D.Sabatini，1978，Proc。娜塔莉。阿卡德。SCI。美国，75：5071-5075)。融合单层细胞经胰酶-EDTA解离后，在旋转培养上清液中培养12~20h后，MDCK细胞失去其特有的结构极性。此外，当这些细胞感染流感或VSV时，病毒粒子以非极化的方式聚集在大部分细胞表面。然而，当将悬浮感染的分离的MDCK细胞稀疏地接种在胶原凝胶上以防止细胞间接触和连接形成时，融合单层中观察到的病毒出芽的特征极性再次显现，即VSV优先从贴壁表面出芽，而流感病毒几乎完全从自由表面出芽。在旋转培养液中培养的聚集细胞中也观察到了类似的极化：流感病毒从自由表面萌发，而VSV则在细胞与细胞接触的区域产生。因此，在分离的上皮细胞中，附着于一种底物或另一种细胞足以触发质膜极性的表达，这体现在病毒的不对称萌发上。
In confluent monolayers of the dog kidney epithelial cell line Madin- Darby canine kidney (MDCK) assembly of RNA enveloped viruses reflects the functional polarization of the cells. Thus, influenza, Sendai, and Simian virus 5 bud from the apical (free) surface, while vesicular stomatitis virions (VSV) are assembled at basolateral plasma membrane domains (Rodriguez-Boulan, E., and D.D. Sabatini, 1978, Proc. Natl. Acad. Sci. U.S.A., 75:5071-5075). MDCK cells derived from confluent monolayers by dissociation with trypsin-EDTA and maintained as single cells in spinner medium for 12-20 h before infection, lose their characteristic structural polarity. Furthermore, when these cells were infected with influenza or VSV, virions assembled in a nonpolarized fashion over most of the cell surface. However, when dissociated MDCK cells infected in suspension were sparsely plated on collagen gels to prevent intercellular contact and the formation of junctions, the characteristic polarity of viral budding observed in confluent monolayers was again manifested; i.e., VSV budded preferentially from adherent surfaces and influenza almost exclusively from free surface regions. Similar polarization was observed in cells which became aggregated during incubation in spinner medium: influenza budded from the free surface, while VSV was produced at regions of cell-cell contact. It therefore appears that in isolated epithelial cells attachment to a substrate or to another cell is sufficient to trigger the expression of plasma membrane polarity which is manifested in the asymmetric budding of viruses.