Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus
Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus
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DOI：
10.1128/jvi.02780-06
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发表时间：
2007-07-01
影响因子：
5.4
通讯作者：
Davey, Robert A.
中科院分区：
文献类型：
--
作者：
Kolokoltsov, Andrey A.;Deniger, Drew;Davey, Robert A.
呼吸道合胞病毒（RSV）是婴儿和老年人呼吸道感染的常见原因。与许多其他非pH依赖性包膜病毒一样，RSV被认为是在细胞表面进入，不依赖于常见的内吞途径。我们使用靶向小干扰RNA（siRNA）文库来鉴定参与细胞骨架动力学和内体运输的关键细胞基因，这些基因对RSV感染很重要。令人惊讶的是，RSV感染被靶向与网格蛋白介导的内吞作用相关的基因（包括网格蛋白轻链）的siRNA有效抑制。网格蛋白介导的内吞作用的重要作用，证实了良好的特点，显性负突变体的基因在这一途径中的表达，并通过使用网格蛋白内吞作用抑制剂氯丙嗪。我们的结论是，虽然RSV可能有能力进入细胞表面，网格蛋白功能和内吞作用是一个生产性RSV感染的必要和重要的组成部分，即使感染是严格独立的pH值。这些研究结果提出的可能性，其他pH值无关的病毒可能会分享一个类似的依赖内吞作用的感染，并提供了一个新的潜在途径治疗感染。
Respiratory syncytial virus (RSV) is a common cause of respiratory tract infections in infants and the elderly. Like many other pH-independent enveloped viruses, RSV is thought to enter at the cell surface, independently of common endocytic pathways. We have used a targeted small interfering RNA (siRNA) library to identify key cellular genes involved in cytoskeletal dynamics and endosome trafficking that are important for RSV infection. Surprisingly, RSV infection was potently inhibited by siRNAs targeting genes associated with clathrin-mediated endocytosis, including clathrin light chain. The important role of clathrin-mediated endocytosis was confirmed by the expression of well-characterized dominant-negative mutants of genes in this pathway and by using the clathrin endocytosis inhibitor chlorpromazine. We conclude that, while RSV may be competent to enter at the cell surface, clathrin function and endocytosis are a necessary and important part of a productive RSV infection, even though infection is strictly independent of pH. These findings raise the possibility that other pH-independent viruses may share a similar dependence on endocytosis for infection and provide a new potential avenue for treatment of infection.