Structural Studies of the T7 DNA Replisome

描述：（由申请人提供）基因组的稳定性， 依赖于忠实的DNA复制，对于正常的细胞功能至关重要。 大多数人类癌症已被证明与某些形式的遗传有关。 不稳定性因此，了解DNA复制的机制是 理解DNA不稳定性的根本原因 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。 本研究的目的是研究全球组织的一个 复制叉，作为理解 在DNA合成过程中， 复制的噬菌体T7 DNA复制系统将被用作 模型系统在这项研究中。电子冷冻显微镜方法将是 用于确定复制体之间形成的复合物的结构， proteins.这项建议的具体目标是： 1.确定T7基因4解旋酶/引发酶和T7 DNA的结构 在前导链和滞后链的背景下的聚合酶复合物 合成，分别使用电子低温显微镜（cryoEM）方法。 2.使用cryoEM确定整个T7噬菌体复制体的结构 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.