Cellular endosomal potassium ion flux regulates arenavirus uncoating during virus entry
Cellular endosomal potassium ion flux regulates arenavirus uncoating during virus entry
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细胞内体钾离子流调节病毒进入过程中沙粒病毒脱壳
DOI：
10.1101/2023.06.23.546275
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Shaw A
中科院分区：
文献类型：
--
作者：
Shaw A
淋巴细胞性脉络丛脑膜炎病毒（LCMV）是布尼亚病毒目沙粒病毒科的一种有囊膜和分节段的负义RNA病毒。LCMV与免疫功能低下人群中的致命性疾病相关，并且作为原型沙粒病毒成员，作为沙粒病毒科的许多高致病性成员的模型，如Junín，Lassa和Lujo病毒，所有这些病毒都与毁灭性出血热相关。为了进入细胞，LCMV包膜与晚期内体膜融合，其中两个既定的要求是低pH值和LCMV糖蛋白（GP）刺突与二级受体CD 164之间的相互作用。LCMV随后脱壳，其中RNA基因组相关核蛋白（NP）与Z蛋白基质层分离，将病毒基因组释放到胞质溶胶中。为了进一步检查LCMV内体逃逸，我们进行了siRNA筛选，其鉴定宿主细胞钾离子（K+）通道对于LCMV感染是重要的，药理学抑制证实了在LCMV进入阶段期间K+通道参与完全消除了生产性感染。为了更好地理解K+介导的感染阻断，我们在生理条件下沿着它们的进入途径追踪进入的病毒体沿着，其中通过分离NP和Z蛋白来表示未包被。相比之下，K+通道阻断阻止了脱壳，将病毒粒子捕获在Rab 7和CD 164阳性内体内，将K+鉴定为第三个LCMV进入要求。K+不增加GP-CD 164结合或改变GP-CD 164依赖性融合。因此，我们建议，K+介导的病毒粒子内部的NP-Z相互作用的调制脱壳。这些结果表明K+通道是一个潜在的抗沙粒病毒的靶点。重要提示沙粒病毒可引起致命的人类疾病，目前尚无有效的预防或治疗方法。在这里，使用原型LCMV，我们确定K+通道对沙粒病毒感染至关重要，在感染周期的进入阶段起着至关重要的作用。我们发现，阻断K+通道功能导致LCMV颗粒被截留在晚期内体隔室中，从而阻止生产性复制。我们的数据表明K+是LCMV脱壳和基因组释放所需的，通过调节病毒核蛋白和病毒颗粒内的基质蛋白层之间的相互作用。
Lymphocytic choriomeningitis virus (LCMV) is an enveloped and segmented negative-sense RNA virus classified within theArenaviridaefamily of theBunyaviralesorder. LCMV is associated with fatal disease in immunocompromised populations and, as the prototypical arenavirus member, acts as a model for the many highly pathogenic members of theArenaviridaefamily, such as Junín, Lassa, and Lujo viruses, all of which are associated with devastating hemorrhagic fevers. To enter cells, the LCMV envelope fuses with late endosomal membranes, for which two established requirements are low pH and interaction between the LCMV glycoprotein (GP) spike and secondary receptor CD164. LCMV subsequently uncoats, where the RNA genome-associated nucleoprotein (NP) separates from the Z protein matrix layer, releasing the viral genome into the cytosol. To further examine LCMV endosome escape, we performed an siRNA screen which identified host cell potassium ion (K+) channels as important for LCMV infection, with pharmacological inhibition confirming K+channel involvement during the LCMV entry phase completely abrogating productive infection. To better understand the K+-mediated block in infection, we tracked incoming virions along their entry pathway under physiological conditions, where uncoating was signified by separation of NP and Z proteins. In contrast, K+channel blockade prevented uncoating, trapping virions within Rab7 and CD164-positive endosomes, identifying K+as a third LCMV entry requirement. K+did not increase GP-CD164 binding or alter GP-CD164-dependent fusion. Thus, we propose that K+mediates uncoating by modulating NP-Z interactions within the virion interior. These results suggest K+channels represent a potential anti-arenaviral target.IMPORTANCEArenaviruses can cause fatal human disease for which approved preventative or therapeutic options are not available. Here, using the prototypical LCMV, we identified K+channels as critical for arenavirus infection, playing a vital role during the entry phase of the infection cycle. We showed that blocking K+channel function resulted in entrapment of LCMV particles within late endosomal compartments, thus preventing productive replication. Our data suggest K+is required for LCMV uncoating and genome release by modulating interactions between the viral nucleoprotein and the matrix protein layer inside the virus particle.
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