Apical recycling systems regulate directional budding of respiratory syncytial virus from polarized epithelial cells
Apical recycling systems regulate directional budding of respiratory syncytial virus from polarized epithelial cells
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DOI：
10.1073/pnas.2434327100
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发表时间：
2003-12-09
影响因子：
11.1
通讯作者：
Crowe, JE
中科院分区：
文献类型：
--
作者：
Brock, SC;Goldenring, JR;Crowe, JE
呼吸道合胞病毒（RSV）是全球婴幼儿严重下呼吸道疾病的主要病毒病因。RSV感染仅限于免疫活性个体的呼吸道上皮的浅表层。与这种体内观察一致，我们和其他人发现RSV优先从受感染的极化上皮细胞的顶面出芽。相反，在非极化的人类上皮细胞中没有观察到定向出芽。这些发现表明RSV使用特定的细胞运输途径来完成病毒复制。调节RSV定向出芽的宿主细胞蛋白是不确定的。极化上皮细胞中细胞蛋白的顶端分选涉及顶端再循环内体（ARE）。为了研究ARE介导的蛋白分选是否在RSV复制过程中发挥作用，我们表达了肌球蛋白Vb尾的片段，其在极化的Madin-Darby犬肾细胞中作为ARE介导的蛋白分选的显性负抑制剂发挥作用。当这些细胞被RSV感染时，观察到病毒产量减少> 9，000倍。当另一个ARE相关蛋白的羧基末端片段Rab 11家族相互作用蛋白1在Madin-Darby犬肾细胞中表达时，观察到对病毒复制的类似影响。这些数据表明，RSV需要适当的ARE介导的蛋白分选，以有效地从极化上皮细胞的顶端表面流出。
Respiratory syncytial virus (RSV) is the major viral cause of serious lower respiratory tract illness in infants and young children worldwide. RSV infection is limited to the superficial layers of the respiratory epithelium in immunocompetent individuals. Consistent with this in vivo observation, we and others have found that RSV buds preferentially from the apical surface of infected polarized epithelial cells. In contrast, directional budding is not observed in nonpolarized human epithelial cells. These findings suggest that RSV uses specific cellular trafficking pathways to accomplish viral replication. The host cell proteins that regulate directional budding of RSV are undefined. Apical sorting of cellular proteins in polarized epithelial cells involves the apical recycling endosome (ARE). To investigate whether ARE-mediated protein sorting plays a role during RSV replication, we expressed a fragment of the myosin Vb tail that functions as a dominant negative inhibitor of ARE-mediated protein sorting in polarized Madin-Darby canine kidney cells. When these cells were infected with RSV, a >9,000-fold reduction in viral yield was observed. A similar effect on virus replication was observed when a carboxyl-terminal fragment of another ARE-associated protein, the Rab11 family interacting protein 1, was expressed in Madin-Darby canine kidney cells. These data suggest that RSV requires proper ARE-mediated protein sorting for efficient egress from the apical surface of polarized epithelial cells.