Watanabe S, Takada A, Watanabe T, Ito H, Kida H, Kawaoka Y
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
J Virol. 2000 Nov;74(21):10194-201. doi: 10.1128/jvi.74.21.10194-10201.2000.
Ebola virus contains a single glycoprotein (GP) that is responsible for receptor binding and membrane fusion and is proteolytically cleaved into disulfide-linked GP1 and GP2 subunits. The GP2 subunit possesses a coiled-coil motif, which plays an important role in the oligomerization and fusion activity of other viral GPs. To determine the functional significance of the coiled-coil motif of GP2, we examined the effects of peptides corresponding to the coiled-coil motif of GP2 on the infectivity of a mutant vesicular stomatitis virus (lacking the receptor-binding/fusion protein) pseudotyped with the Ebola virus GP. A peptide corresponding to the C-terminal helix reduced the infectivity of the pseudotyped virus. We next introduced alanine substitutions into hydrophobic residues in the coiled-coil motif to identify residues important for GP function. None of the substitutions affected GP oligomerization, but some mutations, two in the N-terminal helix and all in the C-terminal helix, reduced the ability of GP to confer infectivity to the mutant vesicular stomatitis virus without affecting the transport of GP to the cell surface, its incorporation into virions, and the production of virus particles. These results indicate that the coiled-coil motif of GP2 plays an important role in facilitating the entry of Ebola virus into host cells and that peptides corresponding to this region could act as efficient antiviral agents.
埃博拉病毒含有一种单一糖蛋白(GP),该糖蛋白负责受体结合和膜融合,并被蛋白水解切割成二硫键连接的GP1和GP2亚基。GP2亚基具有卷曲螺旋基序,这在其他病毒糖蛋白的寡聚化和融合活性中起重要作用。为了确定GP2卷曲螺旋基序的功能意义,我们研究了与GP2卷曲螺旋基序对应的肽对用埃博拉病毒GP假型化的突变水泡性口炎病毒(缺乏受体结合/融合蛋白)感染性的影响。对应于C末端螺旋的肽降低了假型病毒的感染性。接下来,我们将丙氨酸取代引入卷曲螺旋基序中的疏水残基,以鉴定对GP功能重要的残基。没有一个取代影响GP寡聚化,但一些突变,N末端螺旋中的两个突变以及C末端螺旋中的所有突变,降低了GP赋予突变水泡性口炎病毒感染性的能力,而不影响GP向细胞表面的转运、其整合到病毒颗粒中以及病毒颗粒的产生。这些结果表明,GP2的卷曲螺旋基序在促进埃博拉病毒进入宿主细胞中起重要作用,并且对应于该区域的肽可以作为有效的抗病毒剂。