Guo Huichen, Huang Mei, Yuan Quan, Wei Yanquan, Gao Yuan, Mao Lejiao, Gu Lingjun, Tan Yong Wah, Zhong Yanxin, Liu Dingxiang, Sun Shiqi
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping, Lanzhou, Gansu, The P.R. China.
School of Biological Sciences, Nanyang Technological University, Singapore, Singapore.
PLoS One. 2017 Jan 12;12(1):e0170123. doi: 10.1371/journal.pone.0170123. eCollection 2017.
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.
脂筏是包括冠状病毒感染性支气管炎病毒(IBV)在内的一些病毒进入细胞的重要元件。然而,在IBV进入宿主细胞过程中脂筏在细胞膜上的确切作用仍不清楚。在本研究中,我们在用甲基-β-环糊精或美伐他汀耗尽质膜胆固醇后,通过蔗糖密度梯度离心对感染IBV的细胞进行了生物化学分级分离。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物小窝蛋白-1共定位。Vero细胞的感染性测定结果表明,药物诱导的脂筏破坏显著抑制了IBV感染。进一步研究表明,后期IBV基因组复制或病毒粒子释放不需要脂筏。然而,在IBV附着前用药物介导耗尽Vero细胞中的脂筏显著降低了病毒结构蛋白的表达,这表明药物处理损害了IBV与细胞表面的附着。我们的结果表明,脂筏在IBV感染早期,尤其是在附着阶段,作为附着因子发挥作用。