Characterization and disruption of host protein interations required for budding

被包裹的病毒是由萌芽过程形成的，这通常需要宿主蛋白质的参与。 逆转录病毒、横纹病毒和丝状病毒都使用病毒蛋白中类似的晚期结构域序列来 为萌芽招募寄主因子。尽管副粘病毒通常缺乏相同的晚期结构域序列 被这些病毒使用，我们已经获得了证据表明，副粘病毒，类似于其他被囊膜的病毒， 可能会为萌芽招募寄主因素。病毒蛋白：病毒过程中使用的宿主蛋白结合界面 萌芽状态有可能成为抗病毒药物设计的有效靶点。 亨德拉和尼帕病毒(亨尼帕病毒)是最近出现的一种致命的人畜共患副粘病毒。 对人类来说。尽管在表征这些病毒的入侵机制方面取得了进展， 人们对亨尼帕病毒萌发的机制知之甚少。我们打算利用我们的 在副粘病毒萌发领域的专门知识，为此开展实验，目的如下： 1.定义亨尼帕病毒颗粒生产的要求。我们已经建立了类似病毒的颗粒 Hendra和Nipah病毒的(VLP)组装系统。我们将定义哪些亨尼帕病毒蛋白是 对VLP生产很重要，因此哪些蛋白质可能在招募宿主因子方面发挥作用 萌芽中。 2.确定与海尼帕病毒萌发有关的寄主因子。将采用两个独立的战略： 海尼帕病毒VLP宿主蛋白的共亲和纯化及酵母双杂交筛选。一个基于siRNA的基因 二次筛选将确定哪些候选结合蛋白对萌发实际上是重要的。 3.确定抗病毒药物开发的宿主和病毒靶点。病毒和病毒之间的结合界面 宿主蛋白质将被绘制成图谱。将测试宿主蛋白的最小结合片段的阻断能力 病毒萌发，类似于类似片段对逆转录病毒和副流感病毒5(PIV5)的影响 萌芽中。

Enveloped viruses are formed by a budding process that often requires participation of host proteins. Retroviruses, rhabdoviruses, and filoviruses all use similar late domain sequences within the viral proteins to recruit host factors for budding. Although paramyxoviruses generally lack the same late domain sequences used by these viruses, we have obtained evidence that paramyxoviruses, similar to other enveloped viruses, likely recruit host factors for budding. The viral protein:host protein binding interfaces used during virus budding have the potential to be effective as targets for antiviral drug design. Hendra and Nipah viruses (Henipaviruses) are recently emerged, zoonotic paramyxoviruses that are deadly to humans. Although progress has been made in characterizing the entry mechanisms for these viruses, very little is known about mechanisms of Henipavirus budding. We propose to take advantage of our expertise in the field of paramyxovirus budding by conducting experiments with the following aims: 1. Define requirements for Henipavirus particle production. We have established virus-like particle (VLP) assembly systems for Hendra and Nipah virus. We will define which Henipavirus proteins are important for VLP production, and hence which proteins may play roles in the recruitment of host factors for budding. 2. Identify host factors involved in Henipavirus budding. Two independent strategies will be employed: co-affinity purification of host proteins from Henipavirus VLPs, and yeast two-hybrid screening. An siRNAbased secondary screen will determine which candidate binding proteins are actually important for budding. 3. Define host and viral targets for antiviral drug development. Binding interfaces between viral and host proteins will be mapped. Minimal binding fragments of host proteins will be tested for the ability to block virus budding, similar to the effect analogous fragments have on retrovirus and parainfluenza virus 5 (PIV5) budding.