Ebola virus VP40 late domains are not essential for viral replication in cell culture
Ebola virus VP40 late domains are not essential for viral replication in cell culture
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DOI：
10.1128/jvi.79.16.10300-10307.2005
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发表时间：
2005-08-01
影响因子：
5.4
通讯作者：
Kawaoka, Y
中科院分区：
文献类型：
--
作者：
Neumann, G;Ebihara, H;Kawaoka, Y
埃博拉病毒颗粒的形成和出芽是由 VP40 蛋白介导的，该蛋白具有重叠的 PTAP 和 PPXY 晚期结构域基序 (7-PTAPPXY-13)。这些晚期结构域基序也在逆转录病毒的 Gag 蛋白以及弹状病毒和沙粒病毒的基质蛋白中发现。虽然体外研究表明晚期结构域基序在包括埃博拉病毒在内的这些病毒的萌芽中发挥着关键作用，但目前尚不清楚 VP40 晚期结构域是否在埃博拉病毒复制中发挥作用。两个晚期结构域基序的改变显着减少了体外 VP40 颗粒的形成。然而，利用反向遗传学，我们能够生成在一个或两个晚期结构域中含有突变的重组埃博拉病毒。在其一个或两个晚期结构域基序中含有突变的病毒被减毒1个对数单位。透射和扫描电子显微镜没有显示出从受感染细胞释放的突变型病毒和野生型病毒之间存在明显差异。这些发现表明，埃博拉 VP40 晚期结构域基序增强了病毒复制，但并不是细胞培养中病毒复制所必需的。
Ebola virus particle formation and budding are mediated by the VP40 protein, which possesses overlapping PTAP and PPXY late domain motifs (7-PTAPPXY-13). These late domain motifs have also been found in the Gag proteins of retroviruses and the matrix proteins of rhabdo- and arenaviruses. While in vitro studies suggest a critical role for late domain motifs in the budding of these viruses, including Ebola virus, it remains unclear as to whether the VP40 late domains play a role in Ebola virus replication. Alteration of both late domain motifs drastically reduced VP40 particle formation in vitro. However, using reverse genetics, we were able to generate recombinant Ebola virus containing mutations in either or both of the late domains. Viruses containing mutations in one or both of their late domain motifs were attenuated by one log unit. Transmission and scanning electron microscopy did not reveal appreciable differences between the mutant and wild-type viruses released from infected cells. These findings indicate that the Ebola VP40 late domain motifs enhance virus replication but are not absolutely required for virus replication in cell culture.