The biogenesis of a paramyxovirus

我的长期目标是了解病毒的生物发生，探索病毒疾病的基本机制，并确定抗病毒治疗的靶点。背景：病毒是引起公众健康和全球经济日益关注的重要病原体。病毒进入、组装和离开宿主细胞的基本机制对于理解病毒生物学和揭示宿主细胞必不可少的潜在过程至关重要。我已经建造了一个分辨率为10纳米的单分子定位显微镜(SMLM)，大约是蛋白质分子的大小。这款显微镜通过商业上不可用的稳定系统确保高分辨率成像，该系统可以纠正由意外的样品振动引起的成像伪影。通过用这个显微镜观察宿主细胞上的病毒组装位置，我发现副粘病毒Nipah病毒(Niv)的蛋白质以随机的顺序聚集在一起，而不是一个循序渐进的、受调节的过程。这与HIV等其他病毒不同。NIV可以作为不遵循HIV模式的病毒的模型系统。在这个五年计划中，我将研究新城疫的组装和释放机制。我的假设是，NIV的基质蛋白通过与宿主膜脂类和膜裂变机制的相互作用来驱动NIV的组装和释放。我的短期目标是：1)确定病毒在质膜上组装的空间和时间机制；2)通过分析病毒组装部位和脂筏标记之间的空间分布和相互作用动力学，确定膜脂结构域在病毒组装中的作用；3)通过绘制病毒组装部位膜裂变成分的空间组织和募集动力学图，研究膜裂变机制帮助病毒释放的分子机制。影响：分析病毒和细胞中的精确蛋白质组织可以推翻过去的假设，并在病毒研究方面取得突破性进展。通过这样做，我的工作可能会改变抗病毒策略，并揭示抗病毒疗法的新靶点。这项工作还将导致超分辨率显微镜的技术进步，并培训多学科领域的不同群体的研究生和本科生。

My long term objective is to understand virus biogenesis, explore the fundamental mechanisms of viral diseases and identify anti-viral therapeutic targets. Background: Viruses are important pathogens that cause increasing concerns on public health and global economy. Fundamental mechanisms by which viruses enter, assemble and leave host cells are crucial for understanding viral biology and reveal underlying processes essential for the host cell. I have built a single-molecule localization microscope (SMLM) with a resolution of 10 nm, about the size of a protein molecule. This microscope ensures high resolution imaging by a commercial unavailable stabilization system that can correct the imaging artifacts caused by unintended sample vibrations. By visualizing the virus assembly sites on host cells using this microscope, I found that proteins of a paramyxovirus, Nipah virus (NiV), come together in a random order, and not a stepwise, regulated process. This is different from other viruses, such as HIV. NiV can serve as a model system for viruses that do not follow the HIV pattern. In this five-year program, I will study the mechanisms for NiV assembly and release. My hypothesis is that the matrix protein of NiV drives the assembly and release of NiV by its interactions with host membrane lipids and the membrane fission machinery. My short term objectives are to: 1) determine the spatial and temporal mechanisms of the NiV assembly at the plasma membrane; 2) determine the role of membrane lipid domains in virus assembly by analyzing the spatial distribution and interaction dynamics between the virus assembly sites and lipid raft markers; 3) investigate the molecular mechanisms by which the membrane fission machinery helps virus release by mapping out the spatial organization and recruitment dynamics of the membrane fission components at the virus assembly sites. Impact: Analyzing the precise protein organizations in virus and cells can overturn past assumptions and make groundbreaking advances in virus research. In doing so my work may redirect the antivirus strategies and reveal new targets for the antivirus therapeutics. The work will also lead to technological advances in super-resolution microscopy and training of a diverse group of graduate (4) and undergraduate (5) students in multidisciplinary fields.