A PPxY motif within the VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: Implications for filovirus budding
A PPxY motif within the VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: Implications for filovirus budding
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埃博拉病毒 VP40 蛋白中的 PPxY 矩阵与泛素连接酶存在物理和功能上的相互作用： 丝状病毒出芽的意义
DOI：
10.1073/pnas.250277297
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发表时间：
2000-12-05
影响因子：
11.1
通讯作者：
Hayes, FP
中科院分区：
文献类型：
--
作者：
Harty, RN;Brown, ME;Hayes, FP
VP 40是埃博拉病毒和马尔堡病毒的推定基质蛋白，在其N端具有保守的富含脯氨酸基序（PY基序）。我们证明，VP 40蛋白可以介导其自身从哺乳动物细胞中释放，并且PY基序对于这种自我胞吐（出芽）功能是重要的。此外，我们使用蛋白质配体印迹证明，VP 40的PY基序可以介导与具有I型WW-domain的特定细胞蛋白的相互作用。包括哺乳动物泛素连接酶。Nedd4.破坏VP 40的PY基序的单点突变消除了PY/WW结构域的相互作用。值得注意的是，全长VP 40蛋白显示出与全长Rsp 5（酵母的泛素连接酶和Nedd 4的同源物）在物理和功能上相互作用。在体外泛素化测定中，VP 40蛋白以PY依赖的方式被Rsp 5多泛素化。这些数据表明，埃博拉病毒的VP 40蛋白具有PY基序，该基序在功能上类似于先前分别针对特定逆转录病毒和弹状病毒的Gag和M蛋白所描述的那些基序。最后，这些研究暗示VP 40可能在丝状病毒出芽中起重要作用，并且逆转录病毒出芽。弹状病毒和丝状病毒可以通过类似的机制进行。
VP40, the putative matrix protein of both Ebola and Marburg viruses, possesses a conserved proline-rich motif(PY motif) at its N terminus. We demonstrate that the VP40 protein can mediate its own release from mammalian cells, and that the PY motif is important for this self-exocytosis (budding) function. In addition, we used Western-ligand blotting to demonstrate that the PY motif of VP40 can mediate interactions with specific cellular proteins that have type I WW-domains. including the mammalian ubiquitin ligase. Nedd4. Single point mutations that disrupted the PY motif of VP40 abolished the PY/WW-domain interactions. Significantly, the full-length VP40 protein was shown to interact both physically and functionally with full-length Rsp5, a ubiquitin ligase of yeast and homolog of Nedd4 The VP40 protein was multiubiquitinated by Rsp5 in a PY-dependent manner in an in vitro ubiquitination assay. These data demonstrate that the VP40 protein of Ebola Virus possesses a PY motif that is functionally similar to those described previously for Gag and M proteins of specific retroviruses and rhabdoviruses, respectively. Last, these studies imply that VP40 likely plays an important role in filovirus budding, and that budding of retroviruses. rhabdoviruses, and filoviruses may proceed via analogous mechanisms.