Surface expression of viral glycoproteins is polarized in epithelial cells infected with recombinant vaccinia viral vectors.
Surface expression of viral glycoproteins is polarized in epithelial cells infected with recombinant vaccinia viral vectors.
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病毒糖蛋白的表面表达在重组痘苗病毒载体感染的上皮细胞中呈极化。
DOI：
10.1002/j.1460-2075.1986.tb04204.x
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Moss,B
中科院分区：
文献类型：
--
作者：
Stephens,EB;Compans,RW;Earl,P;Moss,B
在极化的上皮细胞中，通过在细胞表面出芽形成的包膜病毒的成熟位点被限制在特定的膜结构域。使用重组牛痘病毒研究了流感病毒血凝素（HA）、水泡性口炎病毒（VSV）G糖蛋白和Friend鼠白血病病毒（MuLV）gp 70/p15 E在Madin达比犬肾（MDCK）细胞系中的表面表达位点。这些糖蛋白可以在感染后4小时通过MDCK细胞表面的免疫荧光证实。在完整的MDCK单层细胞中，表达HA的牛痘重组体产生了典型的顶端表达的糖蛋白的表面荧光图案。相比之下，用表达VSV-G或MuLV gp 70/p15 E的牛痘重组体感染的细胞仅在用EGTA处理单层以破坏紧密连接时才显示表面荧光，正如基底外侧表面上表达的糖蛋白所预期的那样。免疫铁蛋白标记结合电子显微镜证实，感染HA重组体的MDCK细胞表现出顶端表面的特异性标记，而VSV‐G和MuLV重组体表现出各自的抗原主要位于基底外侧膜上。在完整和EGTA处理的单层细胞上通过[125 I]蛋白A结合试验定量表面表达，证实了牛痘表达HA的顶端定位，并证明95%的VSV‐G和97%的MuLV gp 70/p15 E糖蛋白定位于基底外侧表面。这些结果表明，通常在极化上皮细胞的基底外侧表面成熟的病毒的糖蛋白包含其定向运输到基底外侧质膜所需的所有结构信息。
In polarized epithelial cells, maturation sites of enveloped viruses that form by budding at cell surfaces are restricted to particular membrane domains. Recombinant vaccinia viruses were used to investigate the sites of surface expression in the Madin‐Darby canine kidney (MDCK) cell line of the hemagglutinin (HA) of influenza virus, the G glycoprotein of vesicular stomatitis virus (VSV), and gp70/p15E of Friend murine leukemia virus (MuLV). These glycoproteins could be demonstrated by immunofluorescence on the surfaces of MDCK cells as early as 4 h post‐infection. In intact MDCK monolayers, vaccinia recombinants expressing HA produced a pattern of surface fluorescence typical of an apically expressed glycoprotein. In contrast, cells infected with vaccinia recombinants expressing VSV‐G or MuLV gp70/p15E exhibited surface fluorescence only when monolayers were treated with EGTA to disrupt tight junctions, as expected of glycoproteins expressed on basolateral surfaces. Immunoferritin labeling in conjunction with electron microscopy confirmed that MDCK cells infected with the HA recombinant exhibited specific labeling of the apical surfaces whereas the VSV‐G and MuLV recombinants exhibited the respective antigens predominantly on the basolateral membranes. Quantitation of surface expression by [125I]protein A binding assays on intact and EGTA‐treated monolayers confirmed the apical localization of the vaccinia‐expressed HA and demonstrated that 95% of the VSV‐G and 97% of the MuLV gp70/p15E glycoproteins were localized on the basolateral surfaces. These results demonstrate that glycoproteins of viruses that normally mature at basolateral surfaces of polarized epithelial cells contain all of the structural information required for their directional transport to basolateral plasma membranes.