MODULATION OF GLYCOSYLATION AND TRANSPORT OF VIRAL MEMBRANE-GLYCOPROTEINS BY A SODIUM IONOPHORE
MODULATION OF GLYCOSYLATION AND TRANSPORT OF VIRAL MEMBRANE-GLYCOPROTEINS BY A SODIUM IONOPHORE
复制标题
DOI：
10.1083/jcb.97.3.659
复制
发表时间：
1983-01-01
影响因子：
7.8
通讯作者：
COMPANS, RW
中科院分区：
文献类型：
--
作者：
ALONSOCAPLEN, FV;COMPANS, RW
利用荧光活化细胞分选仪（FACS）分析莫能菌素处理细胞表面的病毒糖蛋白表达，结果表明，钠离子载体完全抑制（Madin-Darby犬肾）MDCK细胞表面的水疱性口炎病毒（VSV） G蛋白的出现。观察到流感病毒血凝素（HA）糖蛋白在MDCK细胞表面高水平表达，且其出现的时间过程不受电离层的影响。在VSV或流感病毒感染的细胞中，莫能菌素均未抑制病毒蛋白的合成。病毒糖蛋白的电泳迁移被改变，通过凝胶过滤分析蛋白酶衍生的糖肽表明，在莫能菌素处理的细胞中，唾液酸残基添加到VSV G蛋白上的功能受到损害。在离子载体存在的情况下，观察到流感病毒HA中的焦点和半乳糖掺入减少，但未完全加工的HA蛋白被切割，运输到细胞表面并掺入出芽病毒颗粒中。与之前在MDCK细胞单层培养中观察到的莫能菌素对VSV和流感病毒复制的不同影响相反，在悬浮培养中发现高浓度莫能菌素显著降低了这两种病毒的产量，这表明细胞结构可能在决定病毒复制对药物的敏感性方面发挥作用。尼日利亚素是一种促进K离子跨膜运输的离子载体，可阻断流感病毒和VSV在MDCK细胞单层中的复制，这表明离子载体的离子特异性影响它们对包膜病毒复制的作用。
Analysis of viral glycoprotein expression on surfaces of monensin-treated cells using a fluorescence-activated cell sorter (FACS) demonstrated that the sodium ionophore completely inhibited the appearance of the vesicular stomatitis virus (VSV) G protein on (Madin-Darby canine kidney) MDCK cell surfaces. The expression of the influenza virus hemagglutinin (HA) glycoprotein on the surfaces of MDCK cells was observed to occur at high levels, and the time course of its appearance was not altered by the ionophore. Viral protein synthesis was not inhibited by monensin in either VSV- or influenza virus-infected cells. The electrophoretic mobilities of viral glycoproteins were altered, and analysis of pronase-derived glycopeptides by gel filtration indicated that the addition of sialic acid residues to the VSV G protein was impaired in monensin-treated cells. Reduced incorporation of fucose and galactose into influenza virus HA was observed in the presence of the ionophore, but the incompletely processed HA protein was cleaved, transported to the cell surface and incorporated into budding virus particles. In contrast to the differential effects of monensin on VSV and influenza virus replication previously observed in monolayer cultures of MDCK cells, yields of both viruses were found to be significantly reduced by high concentrations of monensin in suspension cultures, indicating that cellular architecture may play a role in determining the sensitivity of virus replication to the drug. Nigericin, an ionophore that facilitates transport of K ions across membranes, blocked the replication of both influenza virus and VSV in MDCK cell monolayers, indicating that the ion specificity of ionophores influences their effect on the replication of enveloped viruses.