Specific association of glycoprotein B with lipid rafts during herpes simplex virus entry
Specific association of glycoprotein B with lipid rafts during herpes simplex virus entry
复制标题
DOI：
10.1128/jvi.77.17.9542-9552.2003
复制
发表时间：
2003-09-01
影响因子：
5.4
通讯作者：
Cohen, GH
中科院分区：
文献类型：
--
作者：
Bender, FC;Whitbeck, JC;Cohen, GH
单纯疱疹病毒（HSV）进入需要糖蛋白D（gD）与细胞受体如疱疹病毒进入介体（HVEM或HveA）或连接蛋白-1（HveC）的相互作用。然而，融合机制仍然不清楚。由于富含胆固醇的细胞膜脂筏参与了其他包膜病毒（如人类免疫缺陷病毒和埃博拉病毒）的进入，我们测试了HSV进入是否类似。Vero细胞和表达HVEM或nectin-1的细胞用胆固醇螯合药物如甲基-β-环糊精或制霉菌素处理，然后暴露于病毒。在所有情况下，以剂量依赖性方式抑制病毒进入，并且通过补充胆固醇，抑制作用完全可逆。为了研究HVEM和nectin-1与脂筏的关联，我们分析了它们是否分配到非离子洗涤剂不溶性糖脂富集膜（DIG）中。有没有组成性协会的受体与DIG。可溶性gD或病毒与细胞的结合没有导致连接蛋白-1与含筏级分的关联。然而，在感染期间，一部分gB而不是gC、gD或gH与DIG相关。类似地，当细胞与截短的可溶性糖蛋白一起孵育时，发现可溶性gB而不是gC与DIG相关。总之，这些数据有利于HSV使用gB快速动员脂筏的模型，脂筏可以作为进入和细胞信号传导的平台。这也表明，gB可能与脂筏相关的细胞分子相互作用。
Herpes simplex virus (HSV) entry requires the interaction of glycoprotein D (gD) with a cellular receptor such as herpesvirus entry mediator (HVEM or HveA) or nectin-1 (HveC). However, the fusion mechanism is still not understood. Since cholesterol-enriched cell membrane lipid rafts are involved in the entry of other enveloped viruses such as human immunodeficiency virus and Ebola virus, we tested whether HSV entry proceeds similarly. Vero cells and cells expressing either HVEM or nectin-1 were treated with cholesterol-sequestering drugs such as methyl-beta-cyclodextrin or nystatin and then exposed to virus. In all cases, virus entry was inhibited in a dose-dependent manner, and the inhibitory effect was fully reversible by replenishment of cholesterol. To examine the association of HVEM and nectin-1 with lipid rafts, we analyzed whether they partitioned into nonionic detergent-insoluble glycolipid-enriched membranes (DIG). There was no constitutive association of either receptor with DIG. Binding of soluble gD or virus to cells did not result in association of nectin-1 with the raft-containing fractions. However, during infection, a fraction of gB but not gC, gD, or gH associated with DIG. Similarly, when cells were incubated with truncated soluble glycoproteins, soluble gB but not gC was found associated with DIG. Together, these data favor a model in which HSV uses gB to rapidly mobilize lipid rafts that may serve as a platform for entry and cell signaling. It also suggests that gB may interact with a cellular molecule associated with lipid rafts.