The Important Role of Lipid Raft-Mediated Attachment in the Infection of Cultured Cells by Coronavirus Infectious Bronchitis Virus Beaudette Strain.
The Important Role of Lipid Raft-Mediated Attachment in the Infection of Cultured Cells by Coronavirus Infectious Bronchitis Virus Beaudette Strain.
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脂筏介导的附着在冠状病毒传染性支气管炎病毒Beaudette菌株感染培养细胞中的重要作用
DOI：
10.1371/journal.pone.0170123
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Sun S
中科院分区：
文献类型：
--
作者：
Guo H;Huang M;Yuan Q;Wei Y;Gao Y;Mao L;Gu L;Tan YW;Zhong Y;Liu D;Sun S
脂筏是一些病毒进入细胞的重要元件，包括冠状病毒、传染性支气管炎病毒(IBV)。然而，在IBV进入宿主细胞的过程中，脂筏在细胞膜上的确切作用仍不清楚。在这项研究中，我们用甲基-β-环糊精或美伐他汀去除质膜胆固醇后，通过蔗糖密度梯度离心法对感染IBV的细胞进行生化分级。我们的结果表明，与IBV非结构蛋白不同，IBV结构蛋白与脂筏标记aveolin-1共定位。Vero细胞的感染力测定结果表明，药物诱导的脂筏破坏显著抑制了IBV的感染。进一步的研究表明，在后期阶段，IBV基因组复制或病毒粒子释放不需要脂筏。然而，药物介导的Vero细胞在IBV附着前脂筏的耗尽显著减少了病毒结构蛋白的表达，提示药物治疗损害了IBV在细胞表面的附着。我们的结果表明，在IBV感染的早期阶段，尤其是在附着阶段，脂筏起到了附着因子的作用。
Lipid raft is an important element for the cellular entry of some viruses, including coronavirus infectious bronchitis virus (IBV). However, the exact role of lipid rafts in the cellular membrane during the entry of IBV into host cells is still unknown. In this study, we biochemically fractionated IBV-infected cells via sucrose density gradient centrifugation after depleting plasma membrane cholesterol with methyl-β-cyclodextrin or Mevastatin. Our results demonstrated that unlike IBV non-structural proteins, IBV structural proteins co-localized with lipid raft marker caveolin-1. Infectivity assay results of Vero cells illustrated that the drug-induced disruption of lipid rafts significantly suppressed IBV infection. Further studies revealed that lipid rafts were not required for IBV genome replication or virion release at later stages. However, the drug-mediated depletion of lipid rafts in Vero cells before IBV attachment significantly reduced the expression of viral structural proteins, suggesting that drug treatment impaired the attachment of IBV to the cell surface. Our results indicated that lipid rafts serve as attachment factors during the early stages of IBV infection, especially during the attachment stage.