Department of Biomedical Sciences, City University of Hong Kong, Hong Kong, China.
Department of Biomedical Sciences, City University of Hong Kong, Hong Kong, China; Jockey Club of School of Public Health, Chinese University of Hong Kong, Hong Kong, China.
Antiviral Res. 2018 Feb;150:39-46. doi: 10.1016/j.antiviral.2017.11.020. Epub 2017 Nov 24.
Enterovirus A71 (EV-A71) is a small positive-stranded RNA virus that causes human hand, foot and mouth disease (HFMD) and fatal neurological disorders in some cases without effective treatment. Here we show that heat shock cognate protein 70 (Hsc70), a molecular chaperone, displays pivotal role in viral infections. Knockdown of Hsc70 significantly suppresses viral replication evidenced by reducing not only the level of both viral replication intermediates (negative stranded RNA) and viral genomic RNA (positive stranded RNA), but also the level of viral protein expression; whereas ectopic expression of Hsc70 markedly promotes viral replication. Interestingly, depletion of Hsc70 decreases the IRES activity of EV-A71, and the ectopic expression of Hsc70 enhances the IRES activity accordingly. Further study shows that Hsc70 binds viral genomic RNA but does not directly interact with IRES. Moreover, we reveal that Hsc70 interacts with 2A protease and promotes eIF4G cleavage. More importantly, Hsc70 inhibitor Ver-155008 significantly protects cytopathic effects from EV-A71 infection and inhibits both IRES activity and viral reproduction in a dose-dependent manner. The cell viability assay shows that the IC50 and CC50 are 2.01 μM and 47.67 μM, respectively. These results demonstrate not only an important mechanism of Hsc70 in facilitating EV-A71 replication, but also a target for antiviral drug development.
肠道病毒 A71(EV-A71)是一种小的正链 RNA 病毒,可引起人类手足口病(HFMD),并在某些情况下导致致命的神经紊乱,目前尚无有效的治疗方法。在这里,我们表明热休克同源蛋白 70(Hsc70),一种分子伴侣,在病毒感染中发挥关键作用。Hsc70 的敲低显著抑制病毒复制,这表现在不仅降低了两种病毒复制中间体(负链 RNA)和病毒基因组 RNA(正链 RNA)的水平,还降低了病毒蛋白表达的水平;而 Hsc70 的异位表达则显著促进了病毒的复制。有趣的是,Hsc70 的耗竭降低了 EV-A71 的 IRES 活性,而 Hsc70 的异位表达则相应地增强了 IRES 活性。进一步的研究表明,Hsc70 与病毒基因组 RNA 结合,但不直接与 IRES 相互作用。此外,我们揭示 Hsc70 与 2A 蛋白酶相互作用,并促进 eIF4G 的切割。更重要的是,Hsc70 抑制剂 Ver-155008 能显著保护细胞免受 EV-A71 感染的细胞病变效应,并呈剂量依赖性抑制 IRES 活性和病毒复制。细胞活力测定表明,IC50 和 CC50 分别为 2.01 μM 和 47.67 μM。这些结果不仅表明了 Hsc70 在促进 EV-A71 复制中的重要机制,而且还为抗病毒药物的开发提供了一个靶点。