Important role for the transmembrane domain of severe acute respiratory syndrome coronavirus spike protein during entry
Important role for the transmembrane domain of severe acute respiratory syndrome coronavirus spike protein during entry
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DOI：
10.1128/jvi.80.3.1302-1310.2006
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发表时间：
2006-02-01
影响因子：
5.4
通讯作者：
Corver, J
中科院分区：
文献类型：
--
作者：
Broer, R;Boson, B;Corver, J
严重急性呼吸综合征冠状病毒（SARS-CoV）的刺突蛋白（S）负责受体结合和膜融合。它包含一个高度保守的跨膜结构域，该结构域由三部分组成：N末端富含色氨酸的结构域、中心结构域和富含半胱氨酸的C末端结构域。S的胞质尾区先前已被证明是组装所需的。在这里，S的跨膜和胞质结构域在SARS冠状病毒的感染性和膜融合活性的作用进行了研究。SARS-CoV S-假型逆转录病毒（SARSSpp）用于测量S介导的感染性。此外，通过基于海肾淀粉酶的细胞-细胞融合测定来监测S的细胞-细胞融合活性。S-VSV-（Cyt），一种具有来自水泡性口炎病毒G蛋白（VSV-G）的胞质尾的S嵌合体，和SMHV-TMDcyt，一种具有小鼠肝炎病毒胞质和跨膜结构域的S嵌合体，在两种试验中均显示野生型样活性。SVSV-TMDCyt是一种具有VSV-G胞质和跨膜结构域的嵌合体，在SARSpp和细胞-细胞融合试验中受损，与野生型相比显示出3 - 25%的活性，这取决于试验和使用的细胞。SARSpp中嵌合S蛋白的寡聚状态的检查显示SVSV-TMDCyt三聚体比野生型S三聚体稳定性差，这可能解释了融合性和感染性降低。
The spike protein (S) of severe acute respiratory syndrome coronavirus (SARS-CoV) is responsible for receptor binding and membrane fusion. It contains a highly conserved transmembrane domain that consists of three parts: an N-terminal tryptophan-rich domain, a central domain, and a cysteine-rich C-terminal domain. The cytoplasmic tail of S has previously been shown to be required for assembly. Here, the roles of the transmembrane and cytoplasmic domains of S in the infectivity and membrane fusion activity of SARS-CoV have been studied. SARS-CoV S-pseudotyped retrovirus (SARSpp) was used to measure S-mediated infectivity. In addition, the cell-cell fusion activity of S was monitored by a Renilla luciferase-based cell-cell fusion assay. S-VSV-(Cyt), an S chimera with a cytoplasmic tail derived from vesicular stomatitis virus G protein (VSV-G), and SMHV-TMDcyt, an S chimera with the cytoplasmic and transmembrane domains of mouse hepatitis virus, displayed wild-type-like activity in both assays. SVSV-TMDCyt, a chimera with the cytoplasmic and transmembrane domains of VSV-G, was impaired in the SARSpp and cell-cell fusion assays, showing 3 to 25% activity compared to the wild type, depending on the assay and the cells used. Examination of the oligomeric state of the chimeric S proteins in SARSpp revealed that SVSV-TMDCyt trimers were less stable than wild-type S trimers, possibly explaining the lowered fusogenicity and infectivity.