Characterization of rotavirus cell entry
Characterization of rotavirus cell entry
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DOI：
10.1128/jvi.78.5.2310-2318.2004
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发表时间：
2004-03-01
影响因子：
5.4
通讯作者：
López, S
中科院分区：
文献类型：
--
作者：
Martín, CSS;López, T;López, S
虽然最近我们已经了解了很多关于轮状病毒最初附着到MA 104细胞的病毒和细胞蛋白质，但这些病毒到达细胞内部的机制却知之甚少。在这项研究中，我们观察了药物和显性阴性突变体对轮状病毒细胞感染的影响，这些突变体已知会损害网格蛋白介导的内吞作用和小窝介导的内吞作用。轮状病毒能够进入细胞中的化合物，抑制网格蛋白介导的内吞作用，以及细胞过表达的显性负性形式的Eps 15，一种蛋白质的组装网格蛋白外套的存在。我们还发现，轮状病毒感染的细胞，其中小窝摄取被阻断;治疗与胆固醇结合剂制霉菌素和菲律宾，以及转染细胞与显性阴性小窝蛋白-1和小窝蛋白-3突变体，对轮状病毒感染没有影响。有趣的是，用甲基-β-环糊精（一种从膜上隔离胆固醇的药物）处理的细胞，以及表达大的GT3发动蛋白（已知在几种膜断裂事件中起作用）的显性阴性突变体的细胞，没有被轮状病毒感染，表明胆固醇和发动蛋白在轮状病毒的进入中发挥作用。
While recently we have learned much about the viral and cellular proteins involved in the initial attachment of rotaviruses to MA104 cells, the mechanism by which these viruses reach the interior of the cell is poorly understood. For this study, we observed the effects of drugs and of dominant-negative mutants, known to impair clathrin-mediated endocytosis and endocytosis mediated by caveolae, on rotavirus cell infection. Rotaviruses were able to enter cells in the presence of compounds that inhibit clathrin-mediated endocytosis as well as cells overexpressing a dominant-negative form of Eps15, a protein crucial for the assembly of clathrin coats. We also found that rotaviruses infected cells in which caveolar uptake was blocked; treatment with the cholesterol binding agents nystatin and filipin, as well as transfection of cells with dominant-negative caveolin-1 and caveolin-3 mutants, had no effect on rotavirus infection. Interestingly, cells treated with methyl-beta-cyclodextrin, a drug that sequesters cholesterol from membranes, and cells expressing a dominant-negative mutant of the large GTPase dynamin, which is known to function in several membrane scission events, were not infected by rotaviruses, indicating that cholesterol and dynamin play a role in the entry of rotaviruses.