The Orthobunyavirus Germiston Enters Host Cells from Late Endosomes
The Orthobunyavirus Germiston Enters Host Cells from Late Endosomes
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正布尼亚病毒杰米斯顿从晚期内体进入宿主细胞
DOI：
10.1128/jvi.02146-21
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发表时间：
2022
影响因子：
5.4
通讯作者：
P. Lozach
中科院分区：
文献类型：
--
作者：
Stefan Windhaber;Qilin Xin;Zina M. Uckeley;Jana Koch;M. Obr;C. Garnier;Catherine Luengo;Maëva Duboeuf;Florian K. M. Schur;P. Lozach
正布尼亚病毒（obv），包括拉克罗斯病毒、Oropouche病毒和施马伦贝格病毒，对全世界的人类和家畜构成日益严重的威胁。理想情况下，防止OBV传播需要针对感染的早期阶段，即病毒进入的方法。正布尼亚病毒属是包膜RNA病毒的一个大属，在全世界有80多个成员，其中许多是人类和牲畜的新发病原体。正布尼亚病毒（obv）如何穿透和感染哺乳动物宿主细胞仍不清楚。在这里，我们研究了OBV Germiston （GERV）的进入机制。低温电子显微镜观察到病毒颗粒大致呈球形，平均直径为98 nm。用荧光染料标记病毒不会对其传染性产生不利影响，并且可以监测固定细胞和活细胞中的单个颗粒。使用这种方法，我们发现结合病毒的内吞内化是异步的，发生在30到40分钟内。病毒进入Rab5a阳性（Rab5a+）的早期内体，随后进入含有Rab7a但不含LAMP-1的晚期内体空泡。感染进入不需要蛋白水解裂解，内体酸化是充分和必要的病毒融合。酸激活的渗透在病毒内化开始后15 - 25分钟开始，依赖于早期核内体到晚期核内体的成熟。病毒膜融合的最佳pH值略低于6.0，当钾内流被消除时，病毒膜融合的渗透受到阻碍。总的来说，我们的研究提供了GERV进入宿主细胞的实时可视化，并证明了内体成熟晚期对促进OBV渗透的重要性。正布尼亚病毒（obv）包括La Crosse病毒、Oropouche病毒和Schmallenberg病毒，对全世界的人类和家畜构成日益严重的威胁。理想情况下，防止OBV传播需要针对感染的早期阶段，即病毒进入的方法。然而，obv进入并感染宿主细胞的分子和细胞机制尚不清楚。在这里，我们开发了准确、灵敏的工具和检测方法来研究GERV的渗透过程。我们的数据强调了GERV进入后期内体成熟的核心作用，提供了OBV感染早期阶段的全面概述。我们的研究还带来了一个完整的创新方法工具箱来研究OBV进入固定细胞和活细胞的每个步骤，从病毒结合和内吞作用到融合和渗透。本文获得的信息为开发旨在阻止OBV进入的抗病毒策略奠定了基础。
Orthobunyaviruses (OBVs), which include La Crosse, Oropouche, and Schmallenberg viruses, represent a growing threat to humans and domestic animals worldwide. Ideally, preventing OBV spread requires approaches that target early stages of infection, i.e., virus entry. ABSTRACT With more than 80 members worldwide, the Orthobunyavirus genus in the Peribunyaviridae family is a large genus of enveloped RNA viruses, many of which are emerging pathogens in humans and livestock. How orthobunyaviruses (OBVs) penetrate and infect mammalian host cells remains poorly characterized. Here, we investigated the entry mechanisms of the OBV Germiston (GERV). Viral particles were visualized by cryo-electron microscopy and appeared roughly spherical with an average diameter of 98 nm. Labeling of the virus with fluorescent dyes did not adversely affect its infectivity and allowed the monitoring of single particles in fixed and live cells. Using this approach, we found that endocytic internalization of bound viruses was asynchronous and occurred within 30 to 40 min. The virus entered Rab5a-positive (Rab5a+) early endosomes and, subsequently, late endosomal vacuoles containing Rab7a but not LAMP-1. Infectious entry did not require proteolytic cleavage, and endosomal acidification was sufficient and necessary for viral fusion. Acid-activated penetration began 15 to 25 min after initiation of virus internalization and relied on maturation of early endosomes to late endosomes. The optimal pH for viral membrane fusion was slightly below 6.0, and penetration was hampered when the potassium influx was abolished. Overall, our study provides real-time visualization of GERV entry into host cells and demonstrates the importance of late endosomal maturation in facilitating OBV penetration. IMPORTANCE Orthobunyaviruses (OBVs), which include La Crosse, Oropouche, and Schmallenberg viruses, represent a growing threat to humans and domestic animals worldwide. Ideally, preventing OBV spread requires approaches that target early stages of infection, i.e., virus entry. However, little is known about the molecular and cellular mechanisms by which OBVs enter and infect host cells. Here, we developed accurate, sensitive tools and assays to investigate the penetration process of GERV. Our data emphasize the central role of late endosomal maturation in GERV entry, providing a comprehensive overview of the early stages of an OBV infection. Our study also brings a complete toolbox of innovative methods to study each step of the OBV entry program in fixed and living cells, from virus binding and endocytosis to fusion and penetration. The information gained herein lays the foundation for the development of antiviral strategies aiming to block OBV entry.
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加州血清群 Gc (G1) 糖蛋白是 pH 依赖性细胞融合和进入的主要决定因素。
DOI：
10.1016/j.virol.2005.04.026
发表时间：
2005
期刊：
Virology.
影响因子：
--
作者：
Plassmeyer,MatthewL;Soldan,SamanthaS;Stachelek,KarenM;Martin-Garcia,Julio;Gonzalez-Scarano,Francisco
通讯作者：
Gonzalez-Scarano,Francisco
影响因子：
4.5
作者：
Leger, Psylvia;Tetard, Marilou;Lozach, Pierre-Yves
通讯作者：
Lozach, Pierre-Yves
拉克罗斯病毒 G1 的胞外结构域形成寡聚物并经历 pH 依赖性构象变化。
影响因子：
3.7
作者：
Pekosz,A;González-Scarano,F
通讯作者：
González-Scarano,F
影响因子：
5.4
作者：
Meier, Roger;Franceschini, Andrea;Lozach, Pierre-Yves
通讯作者：
Lozach, Pierre-Yves
拉克罗斯病毒 (LACV) Gc 融合肽突变体的生长和融合表型受损，但仍然具有神经毒性。
影响因子：
3.7
作者：
Soldan,SamanthaS;Hollidge,BradleyS;Wagner,Valentina;Weber,Friedemann;Gonzalez-Scarano,Francisco
通讯作者：
Gonzalez-Scarano,Francisco