Use of herpes simplex virus and pseudorabies virus chimeric glycoprotein D molecules to identify regions critical for membrane fusion.
Use of herpes simplex virus and pseudorabies virus chimeric glycoprotein D molecules to identify regions critical for membrane fusion.
复制标题
使用单纯疱疹病毒和伪狂犬病病毒嵌合糖蛋白 D 分子来识别膜融合的关键区域。
DOI：
10.1073/pnas.0408186101
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Spear,PatriciaG
中科院分区：
文献类型：
--
作者：
Zago,Anna;Jogger,CherylR;Spear,PatriciaG
由单纯疱疹病毒（HSV）诱导的膜融合需要四种病毒膜糖蛋白（gB、gD、gH和gL）的作用以及gD与其受体之一（例如疱疹病毒进入介体或nectin-1）的结合。相关的动物疱疹病毒，伪狂犬病病毒（PRV），编码一组同源的糖蛋白，其gD也可以使用nectin-1作为进入受体。我们在这里表明，PRV gD，当与HSV gB，gH和gL共表达时，不能代替HSV gD诱导与表达nectin-1的靶细胞融合。由HSV和PRV序列组成的嵌合gD分子可以取代，条件是前285个氨基酸来自HSV gD。由于前261个氨基酸足以与受体结合，这表明氨基酸262-285含有细胞融合所需的区域，但不含受体结合所需的区域。氨基酸250-299的缺失未能鉴定出对细胞融合至关重要的特定亚区，可能除了氨基酸250-255，其也影响受体结合。相反，膜和受体结合结构域之间的柔性茎的存在似乎是必需的，也许是为了使受体结合和随后的相互作用的gD与膜融合所需的其他糖蛋白的不确定的区域的gD的构象变化。
Membrane fusion induced by herpes simplex virus (HSV) requires the action of four viral membrane glycoproteins (gB, gD, gH, and gL) and the binding of gD to one of its receptors, such as the herpesvirus entry mediator or nectin-1. The related animal herpesvirus, pseudorabies virus (PRV), encodes a homologous set of glycoproteins and its gD can also use nectin-1 as an entry receptor. We show here that PRV gD, when coexpressed with HSV gB, gH, and gL, cannot substitute for HSV gD in inducing fusion with target cells expressing nectin-1. Chimeric gD molecules composed of HSV and PRV sequences can substitute, provided the first 285 aa are from HSV gD. Because the first 261 aa were sufficient for receptor binding, this suggested that amino acids 262–285 contain a region required for cell fusion but not for receptor binding. Deletions from amino acids 250–299 failed to identify a specific subregion critical for cell fusion, except possibly for amino acids 250–255, which also influenced receptor binding. Instead, presence of a flexible stalk between the membrane and receptor-binding domain appears to be required, perhaps to enable conformational changes in gD on receptor binding and subsequent interactions of undefined regions of gD with the other glycoproteins required for membrane fusion.