Structural Studies of the T7 DNA Replisome

描述：(申请人提供)基因组的稳定性，即 依赖于忠实的DNA复制，是正常细胞功能所必需的。 大多数人类癌症已被证明与某些形式的基因有关 不稳定因素。因此，了解DNA复制的机制是 了解与DNA不稳定相关的潜在原因的基础 人类疾病，以及开发潜在的诊断和治疗 这些疾病的治疗方法。 这项研究的目的是研究一个 复制分叉，作为理解 DNA过程中领先链和滞后链的协调合成 复制。噬菌体T7 DNA复制系统将被用作 本研究中的模型系统。电子冷冻显微镜的方法将是 用来确定复制体间形成的复合体的结构 蛋白质。这项建议的具体目标是： 1.确定T7基因4解旋酶/启动酶和T7 DNA的结构 领先链和滞后链背景中的聚合酶复合体 分别用电子冷冻显微镜(CryoEM)方法合成。 2.用低温电子显微镜测定整个T7噬菌体复制体的结构 方法：研究方法。

DESCRIPTION: (provided by applicant) The stability of the genome, which is dependent on faithful DNA replication, is essential to normal cell functions. Most human cancers have been shown to be related to some forms of genetic instabilities. Therefore, understanding the mechanism of DNA replication is fundamental in understanding the underlying causes of DNA instability related human diseases, as well as developing potential diagnostic and therapeutic approaches for these diseases. The objective of this research is to study the global organization of a replication fork, as the first step in understanding the structural basis for the coordinated synthesis of the leading and lagging strands during DNA replication. The bacteriophage T7 DNA replication system will be used as the model system in this study. Electron cryo-microscopic approaches will be employed to determine the structures of the complexes formed between replisomal proteins. The specific aims of this proposal are: 1. Determine the structures of the T7 gene 4 helicase/ primase and the T7 DNA polymerase complexes in the contexts of the leading and lagging strand synthesis, respectively, using electron cryomicroscopic (cryoEM) methods. 2. Determine the structure of the entire T7 phage replisome using cryoEM methods.