Several mammalian arenaviruses (mammarenaviruses) cause hemorrhagic fevers in humans and pose serious public health concerns in their endemic regions. Additionally, mounting evidence indicates that the worldwide-distributed, prototypic mammarenavirus, lymphocytic choriomeningitis virus (LCMV), is a neglected human pathogen of clinical significance. Concerns about human-pathogenic mammarenaviruses are exacerbated by of the lack of licensed vaccines, and current anti-mammarenavirus therapy is limited to off-label use of ribavirin that is only partially effective. Detailed understanding of virus/host-cell interactions may facilitate the development of novel anti-mammarenavirus strategies by targeting components of the host-cell machinery that are required for efficient virus multiplication. Here we document the generation of a recombinant LCMV encoding a nucleoprotein (NP) containing an affinity tag (rLCMV/Strep-NP) and its use to capture the NP-interactome in infected cells. Our proteomic approach combined with genetics and pharmacological validation assays identified ATPase Na+/K+ transporting subunit alpha 1 (ATP1A1) and prohibitin (PHB) as pro-viral factors. Cell-based assays revealed that ATP1A1 and PHB are involved in different steps of the virus life cycle. Accordingly, we observed a synergistic inhibitory effect on LCMV multiplication with a combination of ATP1A1 and PHB inhibitors. We show that ATP1A1 inhibitors suppress multiplication of Lassa virus and Candid#1, a live-attenuated vaccine strain of Junín virus, suggesting that the requirement of ATP1A1 in virus multiplication is conserved among genetically distantly related mammarenaviruses. Our findings suggest that clinically approved inhibitors of ATP1A1, like digoxin, could be repurposed to treat infections by mammarenaviruses pathogenic for humans.

中文翻译：

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感染的细胞中淋巴细胞性脉络膜脑膜炎病毒核蛋白的相互作用研究表明，ATPase Na + / K +转运亚基Alpha 1和禁止素是参与哺乳动物肾炎病毒生命周期的宿主细胞因子。

几种哺乳动物类风疹病毒（哺乳动物肾病毒）会引起人类出血热，并在其流行地区引起严重的公共卫生问题。此外，越来越多的证据表明，全世界分布的原型乳腺肾炎病毒，淋巴细胞性脉络膜脑膜炎病毒（LCMV）是被忽视的具有临床意义的人类病原体。缺乏许可的疫苗加剧了对人致病性丙种肾炎病毒的担忧，并且当前的抗丙种肾病毒治疗仅限于非局部使用利巴韦林，但仅部分有效。通过针对有效病毒繁殖所需的宿主细胞机制组件，对病毒/宿主细胞相互作用的详细了解可以促进新型抗哺乳动物肾病毒策略的开发。在这里，我们记录了编码含有亲和标签（rLCMV / Strep-NP）的核蛋白（NP）的重组LCMV的产生及其在捕获感染细胞中捕获NP-interactome的用途。我们的蛋白质组学方法与遗传学和药理学验证方法相结合，确定了ATPase Na + / K +转运亚基α1（ATP1A1）和禁止素（PHB）为前病毒因子。基于细胞的分析表明，ATP1A1和PHB参与了病毒生命周期的不同步骤。因此，我们观察到与ATP1A1和PHB抑制剂联合使用对LCMV增殖具有协同抑制作用。我们显示ATP1A1抑制剂可抑制Lassa病毒和Candid＃1（Junin病毒的减毒活疫苗株）的繁殖，提示在遗传上远缘相关的哺乳动物肾病毒中，ATP1A1在病毒复制中的需求是保守的。我们的研究结果表明，临床批准的ATP1A1抑制剂（如地高辛）可重新用于治疗对人类致病的哺乳动物肾病毒。