The missing link in coronavirus assembly. Retention of the avian coronavirus infectious bronchitis virus envelope protein in the pre-Golgi compartments and physical interaction between the envelope and membrane proteins.
The missing link in coronavirus assembly. Retention of the avian coronavirus infectious bronchitis virus envelope protein in the pre-Golgi compartments and physical interaction between the envelope and membrane proteins.
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DOI：
10.1074/jbc.m009731200
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发表时间：
2001-05-18
期刊：
影响因子：
--
通讯作者：
Liu DX
中科院分区：
文献类型：
--
作者：
Lim KP;Liu DX
冠状病毒装配中缺少的一个环节是两个关键结构蛋白（膜蛋白（M）和包膜蛋白（E））之间的物理相互作用。在这项研究中，我们证明了冠状病毒传染性支气管炎病毒E可以物理相互作用，通过一个假定的外周结构域，与M。删除该域导致M掺入病毒样颗粒中的急剧减少。共表达M和E的细胞的免疫荧光染色支持E与M相互作用并将M重新定位到E所在的相同亚细胞区室。E被保留在前高尔基体膜中，然后被易位到高尔基体和分泌囊泡;当与E共表达时，观察到M表现出与E相似的定位和易位特征。缺失研究确定C-末端6个残基的RDKLYS是E的内质网滞留信号，并且将−4赖氨酸残基定点突变为谷氨酰胺导致E在高尔基体中积累。在病毒出芽中起关键作用的E的第三个结构域是存在于N-末端区域的推定的跨膜结构域，因为该结构域的缺失导致突变蛋白的自由分布和功能失调的病毒组装。
One missing link in the coronavirus assembly is the physical interaction between two crucial structural proteins, the membrane (M) and envelope (E) proteins. In this study, we demonstrate that the coronavirus infectious bronchitis virus E can physically interact, via a putative peripheral domain, with M. Deletion of this domain resulted in a drastic reduction in the incorporation of M into virus-like particles. Immunofluorescent staining of cells coexpressing M and E supports that E interacts with M and relocates M to the same subcellular compartments that E resides in. E was retained in the pre-Golgi membranes, prior to being translocated to the Golgi apparatus and the secretory vesicles; M was observed to exhibit similar localization and translocation profiles as E when coexpressed with E. Deletion studies identified the C-terminal 6-residue RDKLYS as the endoplasmic reticulum retention signal of E, and site-directed mutagenesis of the −4 lysine residue to glutamine resulted in the accumulation of E in the Golgi apparatus. The third domain of E that plays a crucial role in virus budding is a putative transmembrane domain present at the N-terminal region, because deletion of the domain resulted in a free distribution of the mutant protein and in dysfunctional viral assembly.