Heat shock protein 90 chaperone activity is required for hepatitis A virus replication.

IF 3.8 2区 医学 Q2 VIROLOGY
Journal of Virology Pub Date : 2025-07-22 Epub Date: 2025-06-05 DOI:10.1128/jvi.00502-25
You Li, Xin Zheng, Ling Xie, Maryna Kapustina, Takayoshi Shirasaki, Bryan Yonish, Xian Chen, Asuka Hirai-Yuki, Noriyo Nagata, Ryosuke Suzuki, Masanori Isogawa, Matthew R Vogt, Masamichi Muramatsu, Stanley M Lemon
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引用次数: 0

Abstract

HSP90 heat shock chaperones are essential for maintaining cellular proteostasis, as well as the ATP-dependent folding and functional maturation of many viral proteins. As a result, inhibitors of HSP90 have broad antiviral activity, disrupting replication of many viruses at concentrations below those causing cytotoxicity. Among the Picornaviridae, HSP90 inhibitors block replication of multiple Enterovirus, Aphthovirus, and Cardiovirus species, in some cases, by preventing post-translational processing and assembly of P1 capsid proteins. Hepatitis A virus (HAV), classified within the genus Hepatovirus, has been suggested to be an exception among picornaviruses and to replicate independently of HSP90, possibly because its slow translational kinetics could facilitate co-translational folding and assembly of its capsid proteins. However, we show here that HAV replication is highly dependent upon HSP90, both in human hepatocyte-derived cell lines, in which the 50% inhibitory concentration of geldanamycin was 8.7-11.8 nM, and in vivo in Ifnar1-/- mice. Label-free proteomics experiments suggested that HSP90 interacts with capsid proteins or their precursors and may thus facilitate the folding and assembly of capsid proteins, as it does for enteroviruses and aphthoviruses. By contrast, there was no evidence for HSP90 interacting with any nonstructural protein, and HSP90 inhibitors did not impair 3Cpro proteolytic activity. Despite this, and in contrast to previous studies of enteroviruses and aphthoviruses, geldanamycin potently inhibited replication of a subgenomic HAV replicon. We conclude that HAV is no exception from the HSP90-dependent nature of other picornaviruses and indeed is more dependent on HSP90 than other picornaviruses for amplification of its genome.IMPORTANCEHepatitis A virus (HAV), a common cause of acute infectious hepatitis, has been reported to differ from other picornaviruses in not requiring heat shock protein HSP90 for efficient replication. However, we show here that productive HAV infection is highly dependent on HSP90 and that HAV replication is potently blocked both in cell culture and in vivo in the murine liver by chemical inhibitors of HSP90. Such inhibitors also disrupt the replication of a subgenomic HAV RNA replicon, indicating that HSP90 is required for the assembly of functional replication organelles. This highlights a key difference from other picornaviruses for which HSP90 is required primarily, if not exclusively, for the maturation of the P1 capsid proteins.

热休克蛋白90伴侣活性是甲型肝炎病毒复制所必需的。
HSP90热休克伴侣对于维持细胞蛋白质稳态以及许多病毒蛋白的atp依赖性折叠和功能成熟至关重要。因此,HSP90抑制剂具有广泛的抗病毒活性,在浓度低于引起细胞毒性的浓度时破坏许多病毒的复制。在小核糖核酸病毒科中,HSP90抑制剂通过阻止P1衣壳蛋白的翻译后加工和组装,在某些情况下阻断了多种肠病毒、Aphthovirus和心脏病毒的复制。甲型肝炎病毒(HAV)属于肝病毒属,被认为是小核糖核酸病毒中的一个例外,它可以独立于HSP90复制,可能是因为其缓慢的翻译动力学可以促进其衣壳蛋白的共翻译折叠和组装。然而,我们在这里表明,无论是在人肝细胞来源的细胞系中,格尔达霉素的50%抑制浓度为8.7-11.8 nM,还是在Ifnar1-/-小鼠体内,HAV复制高度依赖于HSP90。无标记蛋白质组学实验表明,HSP90与衣壳蛋白或其前体相互作用,从而可能促进衣壳蛋白的折叠和组装,就像它对肠病毒和阿夫特病毒所做的那样。相比之下,没有证据表明HSP90与任何非结构蛋白相互作用,HSP90抑制剂不会损害3Cpro蛋白水解活性。尽管如此,与之前对肠病毒和口腔病毒的研究相反,格尔达霉素有效地抑制了甲型肝炎亚基因组复制子的复制。我们得出结论,HAV也不例外于其他小核糖核酸病毒的HSP90依赖性,并且在扩增其基因组时确实比其他小核糖核酸病毒更依赖HSP90。甲型肝炎病毒(HAV)是急性感染性肝炎的常见病因,据报道,与其他小核糖核酸病毒不同的是,它不需要热休克蛋白HSP90来进行有效复制。然而,我们在这里表明,HAV感染高度依赖于HSP90,并且HSP90的化学抑制剂在细胞培养和小鼠肝脏中的HAV复制都被有效地阻断。这些抑制剂还会破坏亚基因组HAV RNA复制子的复制,这表明HSP90是功能性复制细胞器组装所必需的。这突出了与其他小核糖核酸病毒的关键区别,其他小核糖核酸病毒的P1衣壳蛋白成熟主要需要HSP90,如果不是唯一的话。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Virology
Journal of Virology 医学-病毒学
CiteScore
10.10
自引率
7.40%
发文量
906
审稿时长
1 months
期刊介绍: Journal of Virology (JVI) explores the nature of the viruses of animals, archaea, bacteria, fungi, plants, and protozoa. We welcome papers on virion structure and assembly, viral genome replication and regulation of gene expression, genetic diversity and evolution, virus-cell interactions, cellular responses to infection, transformation and oncogenesis, gene delivery, viral pathogenesis and immunity, and vaccines and antiviral agents.
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