Characterization and disruption of host protein interations required for budding

出芽所需的宿主蛋白质相互作用的表征和破坏

• 批准号: 8375155
• 负责人: ANTHONY P SCHMITT
• 金额: $ 26.63万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8375155
• 关键词: Affinity Chromatography; Animal Diseases; Antiviral Agents; Binding; Binding Proteins; Development; Drug Delivery Systems; Drug Design; Filovirus; Hendra Virus; Henipavirus; Henipavirus Infections; Human; Integration Host Factors; Maps; Nipah Virus; Paramyxovirus; Play; Probability; Process; Production; Protein Binding; Proteins; Recruitment Activity; Retroviridae; Rhabdoviridae; Role; Screening procedure; System; Testing; Viral; Viral Proteins; Virus; Virus-like particle; Yeasts; biodefense; drug development; human disease; parainfluenza virus; particle; research study; yeast two hybrid system

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.

有包膜病毒是通过出芽过程形成的，通常需要宿主蛋白的参与。 逆转录病毒、弹状病毒和丝状病毒都使用病毒蛋白内相似的晚期结构域序列来 招募出芽宿主因子。尽管副粘病毒通常缺乏相同的晚期结构域序列 这些病毒所使用的，我们已经获得证据表明副粘病毒与其他包膜病毒类似， 可能会招募宿主因子来出芽。病毒蛋白：病毒过程中使用的宿主蛋白结合界面 budding 有潜力作为抗病毒药物设计的有效靶标。 最近出现的亨德拉病毒和尼帕病毒（亨尼帕病毒）是致命的人畜共患副粘病毒 对人类。尽管在描述这些病毒的进入机制方面已经取得了进展， 关于亨尼帕病毒出芽的机制知之甚少。我们建议利用我们的优势 通过进行以下目的的实验，获得副粘病毒领域的专业知识： 1. 明确亨尼帕病毒颗粒生产的要求。我们已经建立了病毒样颗粒 (VLP) 亨德拉病毒和尼帕病毒组装系统。我们将定义哪些亨尼帕病毒蛋白 对于 VLP 的产生很重要，因此哪些蛋白质可能在招募宿主因子中发挥作用 正在萌芽。 2. 确定与亨尼帕病毒出芽有关的宿主因素。将采用两种独立的策略： 亨尼帕病毒 VLP 宿主蛋白的共亲和纯化，以及酵母双杂交筛选。基于 siRNA 的 二次筛选将确定哪些候选结合蛋白对于出芽实际上很重要。 3. 确定抗病毒药物开发的宿主和病毒靶标。病毒与病毒之间的结合界面 宿主蛋白将被映射。将测试宿主蛋白的最小结合片段的阻断能力 病毒出芽，类似于类似片段对逆转录病毒和副流感病毒 5 (PIV5) 的影响 正在萌芽。