Nedd4 regulates egress of Ebola virus-like particles from host cells
Nedd4 regulates egress of Ebola virus-like particles from host cells
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DOI：
10.1128/jvi.77.18.9987-9992.2003
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发表时间：
2003-09-01
影响因子：
5.4
通讯作者：
Shida, H
中科院分区：
文献类型：
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作者：
Yasuda, J;Nakao, M;Shida, H
埃博拉病毒出芽由病毒基质蛋白VP 40内的两个富含脯氨酸的基序PPxY和PTAP介导。我们先前已经表明，Nedd 4样蛋白BUL 1，而不是Nedd 4，正调控D型逆转录病毒Mason-Pfizer猴病毒的出芽（J. Yasuda，E. Hunter，M. Nakao和H. Shida，EMBO代表3：636-640，2002年）。在这里，我们报告说，细胞E3泛素连接酶Nedd 4调节出芽的VP 40诱导的病毒样颗粒（VLP）通过与PPxY基序的相互作用。活性位点半胱氨酸（C894 A）的突变，导致泛素连接酶活性的废除，削弱了Nedd 4出芽的功能。此外，Nedd 4的WW结构域对于结合病毒PPxY基序是必需的，并且仅含有WW结构域的Nedd 4的小片段以显性负性方式显著抑制埃博拉病毒VLP出芽。我们的研究结果表明，含有PPxY作为L-结构域基序的病毒在病毒出芽过程中特异性地使用E3。我们还研究了Tsg 101及其突变体过表达的影响。正如预期的那样，Tsg 101增强了VP 40诱导的VLP释放，而TsgDeltaC，缺乏其C-末端的一半，抑制VLP释放。这些结果表明，Nedd 4与Tsg 101一起在埃博拉病毒出芽中发挥重要作用。
Ebola virus budding is mediated by two proline-rich motifs, PPxY and PTAP, within the viral matrix protein VP40. We have previously shown that a Nedd4-like protein BUL1, but not Nedd4, positively regulates budding of type D retrovirus Mason-Pfizer monkey virus (J. Yasuda, E. Hunter, M. Nakao, and H. Shida, EMBO Rep. 3:636-640, 2002). Here, we report that the cellular E3 ubiquitin ligase Nedd4 regulates budding of VP40-induced virus-like particles (VLPs) through interaction with the PPxY motif. Mutation of the active site cysteine (C894A), resulting in abrogation of ubiquitin ligase activity, impaired the function of Nedd4 on budding. In addition, the WW domains of Nedd4 are essential for binding to the viral PPxY motif, and a small fragment of Nedd4 containing only WW domains significantly inhibited Ebola VLP budding in a dominant-negative manner. Our findings suggest that the viruses containing PPxY as an L-domain motif specifically use E3 in the process of virus budding. We also examined the effects of overexpression of Tsg101 and its mutant. As expected, Tsg101 enhanced VP40-induced VLP release, and TsgDeltaC, which lacks its C-terminal half, inhibited VLP release. These results indicate that Nedd4, together with Tsg101, plays an important role in Ebola virus budding.