Physical and Biochemical Characterization of the Portal Complex of Bacteriophage Lambda

DNA包装是许多双链DNA病毒（包括痘病毒、疱疹病毒和许多噬菌体）组装的关键步骤。在所有情况下，病毒基因组都被终止酶插入预先形成的衣壳中。DNA通过入口进入衣壳，这是一种在所有这些病毒衣壳中发现的甜甜圈形状的结构。这个实验室的长期目标是详细了解病毒基因组包装的过程。本项目检查DNA包装在噬菌体lambda作为一个模型系统，了解病毒组装的基本机制。在病毒衣壳成熟过程中，lambda门户被广泛修饰，本项目的一个主要目标是表征lambda门户的化学和结构修饰，并定义每种修饰的生物学作用。此外，负责衣壳成熟的病毒蛋白酶将被识别和表征。最近出现的“新”病毒祸害，病毒包装系统在纳米科学和工程中的潜力，以及生物武器日益增加的威胁，都强调需要在最基本的层面上了解病毒的发展。这项研究的结果将有助于建立这一重要的知识库，并使对新的和正在出现的生物威胁作出快速反应成为可能。更广泛的影响：博士前和博士后学生，包括在科学领域代表性不足的少数群体成员，将参与这项工作。因此，该项目将有助于培养一批多样化的未来科学家。

DNA packaging is a critical step in the assembly of many double-stranded DNA viruses, including poxvirus, the herpesviruses, and many bacteriophages. In all cases, the viral genome is inserted into a pre-formed capsid by a terminase enzyme. DNA passes into the capsid through the portal, a doughnut-shaped structure found in the capsids of all of these viruses. The long-range goal of this lab is to understand, in detail, the process of viral genome packaging. The present project examines DNA packaging in bacteriophage lambda as a model system to understand the basic mechanisms of virus assembly. The lambda portal is extensively modified during the maturation of the viral capsid, and a major goal of this project is to characterize the chemical and structural modifications of the lambda portal, and to define the biological role of each modification. In addition, the viral protease responsible for capsid maturation will be identified and characterized. The recent emergence of "new" viral scourges, the potential of viral packaging systems in nanosciences and engineering, and the increasing threat of biological weapons underscore the need to understand virus development at the most fundamental level. The results of this research will contribute to this critical knowledge base, and make possible a rapid response to new and emerging biological threats. Broader Impacts: Pre-doctoral and post-doctoral students, including members of minority groups that have been underrepresented in science, will participate in this work. The project will thus serve to train a diverse group of future scientists.