The PDZ3 domain of the cellular scaffolding protein MAGI-1 interacts with the Coxsackievirus and adenovirus receptor (CAR).
The PDZ3 domain of the cellular scaffolding protein MAGI-1 interacts with the Coxsackievirus and adenovirus receptor (CAR).
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细胞支架蛋白 MAGI-1 的 PDZ3 结构域与柯萨奇病毒和腺病毒受体 (CAR) 相互作用。
DOI：
10.1016/j.biocel.2015.01.012
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Excoffon,KatherineJDA
中科院分区：
文献类型：
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作者：
Yan,Ran;Sharma,Priyanka;Kolawole,AbimbolaO;Martin,SterlingCT;Readler,JamesM;Kotha,PoornimaLN;Hostetler,HeatherA;Excoffon,KatherineJDA
柯萨奇病毒和腺病毒受体（CAR）是一种重要的细胞蛋白，参与细胞-细胞粘附、蛋白运输和病毒感染。CAR的主要同种型被选择性地分选到极化上皮细胞的基底外侧膜，在那里它与具有反向结构域结构的细胞支架蛋白膜相关鸟苷酸激酶-1（MAGI-1）共定位。先前，我们证明了CAR通过PDZ结构域依赖性相互作用与MAGI-1相互作用。在这里，我们表明MAGI-1的PDZ 3结构域是唯一负责CAR和MAGI-1的7个外显子异构体之间的高亲和力相互作用，使用酵母双杂交分析，并确认这种相互作用的生物化学和细胞裂解物中通过体外下拉测定和免疫共沉淀。通过荧光共振能量转移测量PDZ 3结构域和CAR C-末端之间的高亲和力相互作用。此外，我们研究了CAR和MAGI-1的PDZ 3结构域之间的这种高亲和力相互作用的生物学相关性，并发现它不改变CAR介导的腺病毒感染。相比之下，这种高亲和力相互作用的中断改变了MAGI-1的定位，表明CAR能够将MAGI-1运输到细胞连接处。这些数据加深了对CAR和MAGI-1之间相互作用的分子理解，并表明尽管CAR在将基于PDZ的支架蛋白运输到细胞连接中起作用，但与高亲和力细胞内结合伴侣的结合并不显著改变腺病毒通过CAR的结合和进入。
The Coxsackievirus and adenovirus receptor (CAR) is an essential cellular protein that is involved in cell–cell adhesion, protein trafficking, and viral infection. The major isoform of CAR is selectively sorted to the basolateral membrane of polarized epithelial cells where it co-localizes with the cellular scaffolding protein membrane-associated guanylate kinase with inverted domain structure-1 (MAGI-1). Previously, we demonstrated CAR interacts with MAGI-1 through a PDZ–domain dependent interaction. Here, we show that the PDZ3 domain of MAGI-1 is exclusively responsible for the high affinity interaction between the seven exon isoform of CAR and MAGI-1 using yeast-two-hybrid analysis and confirming this interaction biochemically and in cellular lysates byin vitropull down assay and co-immunoprecipitation. The high affinity interaction between the PDZ3 domain and CAR C-terminus was measured by fluorescence resonance energy transfer. Further, we investigated the biological relevance of this high affinity interaction between CAR and the PDZ3 domain of MAGI-1 and found that it does not alter CAR-mediated adenovirus infection. By contrast, interruption of this high affinity interaction altered the localization of MAGI-1 indicating that CAR is able to traffic MAGI-1 to cell junctions. These data deepen the molecular understanding of the interaction between CAR and MAGI-1 and indicate that although CAR plays a role in trafficking PDZ-based scaffolding proteins to cellular junctions, association with a high affinity intracellular binding partner does not significantly alter adenovirus binding and entry via CAR.