Biochemical, Biophysical and Structural Characterization of Phage Lambda Capsid Assembly and Maturation

疱疹病毒和许多噬菌体，感染细菌的病毒，有着相似的发育途径。所有这些病毒的一个重要步骤是组装一个保护环境中病毒基因组的衣壳。衣壳由多个拷贝的主要衣壳蛋白和支架“伴侣”蛋白自组装。壳的组装和随后的稳定性所需的相互作用对病毒复制至关重要。该项目研究并定义了这些相互作用，这将促进我们对病毒如何在环境中复制和保持感染性的理解。了解这些力量对于合理开发抗病毒疗法和调整新型纳米材料的结构以用于科学和潜在的健康相关应用也是必要的。在这个项目中进行的工作将由有前途的年轻科学研究人员从本科生到研究生到博士后研究员进行，并将用于培养美国劳动力中的下一代科学家。具体来说，该项目将定义外壳组装和稳定所需的衣壳蛋白之间的结构和热力学相互作用。这些研究将以协调的方式采用生物化学（诱变）、生物物理（分析超离心）、计算（分子动力学）和结构（核磁共振、晶体学、冷冻电子显微镜）方法。还询问了高保真壳组装所需的支架和衣壳蛋白之间的基本相互作用。研究结果将揭示原核和真核大双链DNA病毒中壳组装和稳定性所必需的分子相互作用。这些数据在抗病毒治疗、噬菌体治疗和纳米技术应用方面具有重要意义。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The herpesviruses and many bacteriophages, viruses that infect bacteria, share similar developmental pathways. An essential step for all of them is the assembly of a capsid shell that protects the viral genome in the environment. The capsid self-assembles from multiple copies of a major capsid protein and a scaffolding “chaperone” protein. The interactions required for assembly and subsequent stability of the shell are of fundamental importance to virus replication. This project studies and defines these interactions which will advance our understanding of how the viruses replicate and maintain infectivity in the environment. An understanding of these forces is also necessary for the rational development of anti-viral therapeutics and to adapt the structures for novel nanomaterials for scientific and potentially health-related applications. The work performed in this project will be conducted by promising young scientific investigators from undergraduate to graduate to post-doctoral fellows and will serve to train the next generation of scientists in the American work force.Specifically, the project will define the structural and thermodynamic interactions between the capsid proteins required for shell assembly and stability. The studies will employ biochemical (mutagenesis), biophysical (analytical ultracentrifugation), computational (molecular dynamics) and structural (Nuclear Magnetic Resonance, crystallography, cryo-Electron Microscopy) approaches in a coordinated manner. Fundamental interactions between the scaffolding and capsid proteins required for high-fidelity shell assembly are also interrogated. The results of the studies will reveal molecular interactions essential to shell assembly and stability in the large double-stranded DNA viruses, both prokaryotic and eukaryotic. These data have significant implications in anti-viral therapeutics, phage therapy and nanotechnology applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.