Coronavirus-induced membrane fusion requires the cysteine-rich domain in the spike protein.
Coronavirus-induced membrane fusion requires the cysteine-rich domain in the spike protein.
复制标题
DOI：
10.1006/viro.2000.0219
复制
发表时间：
2000-03-30
期刊：
影响因子：
3.7
通讯作者：
Gombold JL
中科院分区：
文献类型：
--
作者：
Chang KW;Sheng Y;Gombold JL
小鼠肝炎病毒A59株的刺突糖蛋白介导了导致细胞感染的早期事件，包括病毒和细胞膜的融合。Spike是一种I型膜糖蛋白，具有一个保守的跨膜锚定和一个罕见的富含半胱氨酸(Cys)的结构域，该结构域连接了锚定和细胞质尾巴的假定连接。在这项研究中，我们研究了这些羧基末端结构域在尖峰信号转导的膜融合中的作用。我们发现，细胞质尾巴不是融合所必需的，但具有增强膜融合活性的能力。含有整个跨膜锚点和CyS结构域与单纯疱疹病毒1型糖蛋白D(GD-1)锚点替换的嵌合Spike蛋白突变体表明，融合活性需要A59跨膜结构域和位于细胞质尾部上游的CyS结构域的存在。Cys结构域是必需的元件，因为它从野生型Spike蛋白中删除会取消融合活性。然而，将半胱氨酸结构域添加到融合缺陷嵌合蛋白中不能恢复融合活性。因此，CyS结构域是必要的，但不足以补充GD-1锚点并允许膜融合。半胱氨酸区保守半胱氨酸残基的定点突变显著减少了膜融合，这进一步支持了该区域对棘波功能至关重要的结论。结果表明，钉状跨膜锚定的羧基末端至少含有两个不同的结构域，这两个结构域都是完全膜融合所必需的。
The spike glycoprotein of mouse hepatitis virus strain A59 mediates the early events leading to infection of cells, including fusion of the viral and cellular membranes. The spike is a type I membrane glycoprotein that possesses a conserved transmembrane anchor and an unusual cysteine-rich (cys) domain that bridges the putative junction of the anchor and the cytoplasmic tail. In this study, we examined the role of these carboxyl-terminal domains in spike-mediated membrane fusion. We show that the cytoplasmic tail is not required for fusion but has the capacity to enhance membrane fusion activity. Chimeric spike protein mutants containing substitutions of the entire transmembrane anchor and cys domain with the herpes simplex virus type 1 glycoprotein D (gD-1) anchor demonstrated that fusion activity requires the presence of the A59 membrane-spanning domain and the portion of the cys domain that lies upstream of the cytoplasmic tail. The cys domain is a required element since its deletion from the wild-type spike protein abrogates fusion activity. However, addition of the cys domain to fusion-defective chimeric proteins was unable to restore fusion activity. Thus, the cys domain is necessary but is not sufficient to complement the gD-1 anchor and allow for membrane fusion. Site-specific mutations of conserved cysteine residues in the cys domain markedly reduce membrane fusion, which further supports the conclusion that this region is crucial for spike function. The results indicate that the carboxyl-terminus of the spike transmembrane anchor contains at least two distinct domains, both of which are necessary for full membrane fusion.
登录
查看更多内容
影响因子：
5.4
作者：
BRODY, BA;RHEE, SS;HUNTER, E
通讯作者：
HUNTER, E
影响因子：
5.4
作者：
Berlioz-Torrent, C;Shacklett, BL;Benarous, R
通讯作者：
Benarous, R
影响因子：
3.8
作者：
GOMBOLD, JL;SUTHERLAND, RM;WEISS, SR
通讯作者：
WEISS, SR
影响因子：
5.4
作者：
GALLAGHER, TM;ESCARMIS, C;BUCHMEIER, MJ
通讯作者：
BUCHMEIER, MJ
影响因子：
7.8
作者：
Bagai, S;Lamb, RA
通讯作者：
Lamb, RA