Structure-function analysis of reovirus binding to junctional adhesion molecule 1 - Implications for the mechanism of reovirus attachment
Structure-function analysis of reovirus binding to junctional adhesion molecule 1 - Implications for the mechanism of reovirus attachment
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DOI：
10.1074/jbc.m305649200
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发表时间：
2003-11-28
影响因子：
4.8
通讯作者：
Dermody, TS
中科院分区：
文献类型：
--
作者：
Forrest, JC;Campbell, JA;Dermody, TS
哺乳动物呼肠孤病毒是具有长的丝状附着蛋白的无包膜病毒，该蛋白决定新生小鼠感染后的疾病表型，并且是腺病毒附着蛋白的结构同源物。呼肠孤病毒使用连接粘附分子1（JAM 1）作为非依赖于病毒型的细胞受体。JAM1是一种广泛表达的免疫球蛋白超家族蛋白，可形成稳定的同源二聚体并调节紧密连接通透性和淋巴细胞运输。我们采用了一系列的结构指导的结合和感染实验，以确定在人JAM1（hJAM1）的呼肠孤病毒受体相互作用的重要残基，并深入了解呼肠孤病毒的附着机制。使用嵌合和结构域缺失突变体受体分子的结合和感染实验表明，呼肠孤病毒附着、感染和复制需要hJAM1的氨基末端D1结构域。呼肠孤病毒与hJAM1的结合比同型hJAM1结合更快，并且在体外和细胞上被过量的hJAM1竞争。交联hJAM1降低了呼肠孤病毒在体外和细胞上结合hJAM1的能力，并消除了可溶性hJAM1对呼肠孤病毒附着的竞争作用。最后，诱变研究表明，与hJAM1二聚体界面密切相关的残基是呼肠孤病毒与hJAM1相互作用的关键。这些结果表明，呼肠孤病毒附着破坏hJAM1二聚体，并突出呼肠孤病毒和腺病毒的附着策略之间的相似性。
Mammalian reoviruses are nonenveloped viruses with a long, filamentous attachment protein that dictates disease phenotypes following infection of newborn mice and is a structural homologue of the adenovirus attachment protein. Reoviruses use junctional adhesion molecule 1 (JAM1) as a serotype-independent cellular receptor. JAM1 is a broadly expressed immunoglobulin superfamily protein that forms stable homodimers and regulates tight-junction permeability and lymphocyte trafficking. We employed a series of structure-guided binding and infection experiments to define residues in human JAM1 (hJAM1) important for reovirus-receptor interactions and to gain insight into mechanisms of reovirus attachment. Binding and infection experiments using chimeric and domain deletion mutant receptor molecules indicate that the amino-terminal D1 domain of hJAM1 is required for reovirus attachment, infection, and replication. Reovirus binding to hJAM1 occurs more rapidly than homotypic hJAM1 association and is competed by excess hJAM1 in vitro and on cells. Crosslinking hJAM1 diminishes the capacity of reovirus to bind hJAM1 in vitro and on cells and negates the competitive effects of soluble hJAM1 on reovirus attachment. Finally, mutagenesis studies demonstrate that residues intimately associated with the hJAM1 dimer interface are critical for reovirus interactions with hJAM1. These results suggest that reovirus attachment disrupts hJAM1 dimers and highlight similarities between the attachment strategies of reovirus and adenovirus.