结构表征和 CD2AP 二价结合到基性无序域的 chikungunya 病毒 nsP3 蛋白的生物学功能。
Structural characterization and biological function of bivalent binding of CD2AP to intrinsically disordered domain of chikungunya virus nsP3 protein.
机构信息
Department of Molecular Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Department of Microbiology, University of Alabama at Birmingham, AL, USA.
出版信息
Virology. 2019 Nov;537:130-142. doi: 10.1016/j.virol.2019.08.022. Epub 2019 Aug 22.
Alphavirus nsP3 proteins contain long, intrinsically disordered, hypervariable domains, HVD, which serve as hubs for interaction with many cellular proteins. Here, we have deciphered the mechanism and function of HVD interaction with host factors in alphavirus replication. Using NMR spectroscopy, we show that CHIKV HVD contains two SH3 domain-binding sites. Using an innovative chemical shift perturbation signature approach, we demonstrate that CD2AP interaction with HVD is mediated by its SH3-A and SH3-C domains, and this leaves the SH3-B domain available for interaction with other cellular factor(s). This cooperative interaction with two SH3 domains increases binding affinity to CD2AP and possibly induces long-range allosteric effects in HVD. Our data demonstrate that BIN1, CD2AP and SH3KBP1 play redundant roles in initiation of CHIKV replication. Point mutations in both CHIKV HVD binding sites abolish its interaction with all three proteins, CD2AP, BIN1 and SH3KBP1. This results in strong inhibition of viral replication initiation.
甲病毒 nsP3 蛋白含有长的、固有无序的、高变区（HVD），它作为与许多细胞蛋白相互作用的中心。在这里，我们已经破译了 HVD 与甲病毒复制中宿主因子相互作用的机制和功能。使用 NMR 光谱，我们表明 CHIKV HVD 包含两个 SH3 结构域结合位点。使用创新的化学位移扰动特征方法，我们证明 CD2AP 与 HVD 的相互作用是由其 SH3-A 和 SH3-C 结构域介导的，这使得 SH3-B 结构域可用于与其他细胞因子（s）相互作用。这种与两个 SH3 结构域的协同相互作用增加了与 CD2AP 的结合亲和力，并可能在 HVD 中诱导长程变构效应。我们的数据表明，BIN1、CD2AP 和 SH3KBP1 在 CHIKV 复制的起始中发挥冗余作用。在 CHIKV HVD 的两个结合位点上的点突变都使其丧失与这三种蛋白（CD2AP、BIN1 和 SH3KBP1）的相互作用。这导致病毒复制起始的强烈抑制。
Zotero 插件