The SARS coronavirus E protein interacts with PALS1 and alters tight junction formation and epithelial morphogenesis.
The SARS coronavirus E protein interacts with PALS1 and alters tight junction formation and epithelial morphogenesis.
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DOI：
10.1091/mbc.e10-04-0338
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发表时间：
2010-11-15
影响因子：
3.3
通讯作者：
Nal B
中科院分区：
文献类型：
--
作者：
Teoh KT;Siu YL;Chan WL;Schlüter MA;Liu CJ;Peiris JS;Bruzzone R;Margolis B;Nal B
严重急性呼吸综合征冠状病毒的 E 蛋白通过新型 PDZ 结构域结合基序/PDZ 结构域相互作用结合紧密连接蛋白 PALS1。 E 的表达延迟紧密连接的形成并改变 MDCKII 上皮细胞的囊肿形态发生。细胞间紧密连接定义了上皮顶端基底极性并形成物理围栏，保护下面的组织免受病原体入侵。 PALS1 是一种紧密连接相关蛋白，是 CRUMBS3-PALS1-PATJ 极性复合体的成员，该复合体对于哺乳动物上皮极性的建立和维持至关重要。在这里，我们报告 SARS-CoV E 小包膜蛋白 (E) 的羧基末端结构域与人类 PALS1 结合。使用免疫共沉淀和下拉实验，我们证明 E 与哺乳动物细胞中的 PALS1 相互作用，并进一步证明 E 的最后四个羧基末端氨基酸形成一个新的 PDZ 结合基序，该基序与 PALS1 PDZ 结构域结合。在感染 SARS-CoV 的 Vero E6 细胞中，PALS1 重新分布到 ERGIC/高尔基体区域，E 在此区域积累。 MDCKII 上皮细胞中 E 的异位表达显着改变囊肿形态发生，此外，以 PDZ 结合基序依赖性方式延迟紧密连接的形成、影响极性并改变 PALS1 的亚细胞分布。我们推测 SARS-CoV E 劫持 PALS1 在 SARS 患者肺上皮细胞破坏中起着决定性作用。
The E protein of the Severe Acute Respiratory Syndrome Coronavirus binds the tight junction protein PALS1 through a novel PDZ domain-binding motif/PDZ domain interaction. Expression of E delays formation of tight junctions and alters cyst morphogenesis of MDCKII epithelial cells. Intercellular tight junctions define epithelial apicobasal polarity and form a physical fence which protects underlying tissues from pathogen invasions. PALS1, a tight junction-associated protein, is a member of the CRUMBS3-PALS1-PATJ polarity complex, which is crucial for the establishment and maintenance of epithelial polarity in mammals. Here we report that the carboxy-terminal domain of the SARS-CoV E small envelope protein (E) binds to human PALS1. Using coimmunoprecipitation and pull-down assays, we show that E interacts with PALS1 in mammalian cells and further demonstrate that the last four carboxy-terminal amino acids of E form a novel PDZ-binding motif that binds to PALS1 PDZ domain. PALS1 redistributes to the ERGIC/Golgi region, where E accumulates, in SARS-CoV–infected Vero E6 cells. Ectopic expression of E in MDCKII epithelial cells significantly alters cyst morphogenesis and, furthermore, delays formation of tight junctions, affects polarity, and modifies the subcellular distribution of PALS1, in a PDZ-binding motif-dependent manner. We speculate that hijacking of PALS1 by SARS-CoV E plays a determinant role in the disruption of the lung epithelium in SARS patients.