Coronavirus envelope (E) protein remains at the site of assembly.
Coronavirus envelope (E) protein remains at the site of assembly.
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DOI：
10.1016/j.virol.2015.02.005
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发表时间：
2015-04
期刊：
影响因子：
3.7
通讯作者：
Hogue BG
中科院分区：
文献类型：
--
作者：
Venkatagopalan P;Daskalova SM;Lopez LA;Dolezal KA;Hogue BG
冠状病毒 (CoV) 在内质网高尔基中间室 (ERGIC) 膜上组装，并从货物囊泡中的细胞中排出。只有少数包膜 (E) 蛋白分子被组装成病毒体。 E 在形态发生中的作用尚不完全清楚。研究人员对小鼠肝炎病毒 A59 (MHV) E 蛋白的细胞定位和动态进行了研究，以进一步了解其在感染过程中的作用。 E蛋白定位于ERGIC和高尔基体，氨基末端和羧基末端分别位于管腔和细胞质中。 E 蛋白不运输至细胞表面。 MHV 经过基因改造，在 E 的羧基末端添加了四半胱氨酸标签。光漂白后的荧光恢复 (FRAP) 显示 E 在 ERGIC/高尔基体膜中可移动。相关光电子显微镜 (CLEM) 证实高尔基池中存在 E。这些结果有力地支持了 E 蛋白在出芽/组装位点发挥其功能。小鼠肝炎冠状病毒 (MHV-CoV) E 蛋白定位于 ERGIC 和高尔基体。 MHV-CoV E 不会转运至细胞表面。 MHV-CoV 可以通过在 E 上附加四半胱氨酸标签进行基因工程。首个 FRAP 和 CoV E 蛋白的相关光学电子显微镜。活细胞成像显示 E 在 ERGIC/高尔基体膜中可移动。
Coronaviruses (CoVs) assemble at endoplasmic reticulum Golgi intermediate compartment (ERGIC) membranes and egress from cells in cargo vesicles. Only a few molecules of the envelope (E) protein are assembled into virions. The role of E in morphogenesis is not fully understood. The cellular localization and dynamics of mouse hepatitis CoV A59 (MHV) E protein were investigated to further understanding of its role during infection. E protein localized in the ERGIC and Golgi with the amino and carboxy termini in the lumen and cytoplasm, respectively. E protein does not traffic to the cell surface. MHV was genetically engineered with a tetracysteine tag at the carboxy end of E. Fluorescence recovery after photobleaching (FRAP) showed that E is mobile in ERGIC/Golgi membranes. Correlative light electron microscopy (CLEM) confirmed the presence of E in Golgi cisternae. The results provide strong support that E proteins carry out their function(s) at the site of budding/assembly. Mouse hepatitis coronavirus (MHV-CoV) E protein localizes in the ERGIC and Golgi. MHV-CoV E does not transport to the cell surface. MHV-CoV can be genetically engineered with a tetracysteine tag appended to E. First FRAP and correlative light electron microscopy of a CoV E protein. Live-cell imaging shows that E is mobile in ERGIC/Golgi membranes.