Tn7 Transpostion into DNA Replication/Repair Complexes

Tn7 转座到 DNA 复制/修复复合物中

• 批准号: 7073909
• 负责人: Joseph E Peters
• 金额: $ 22.1万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7073909
• 关键词: DNA binding protein; DNA repair; DNA replication; alanine; bacterial genetics; bacterial proteins; biological signal transduction; mutant; nucleic acid metabolism; plasmids; protein structure function; transposon /insertion element

DESCRIPTION (provided by applicant): The long-term goal of the lab is to understand how organisms maintain their genomic information at the molecular level. We are investigating one of the ways genomes can be altered in a process called transposition. Transposons are mobile DNA elements that can move within a cell which are found in nearly every organism and are abundant in the human genome. To better understand transposition at the molecular level we are using the bacterial transposon Tn7. Tn7 transposition is of particular interest in that it also allows us to understand DNA replication and repair because Tn7 recognizes complexes involved in these processes as insertion targets. This work is related to human health because it will lead to a better understanding of cellular DNA repair functions found in all living organisms that are important in preventing cancer. Tn7 is able to recognize a structure or complex associated with multiple forms of DNA metabolism using the Tn7-encoded protein TnsE. TnsE-mediated transposition preferentially occurs into plasmids capable of moving between cells called conjugal plasmids. The TnsE-mediated pathway also recognizes structures associated with the repair of DNA double-strand breaks and allows Tn7 to preferentially direct transposition where DNA replication terminates. We will determine if TnsE interacts directly with the beta-clamp processivity factor to recognize lagging- strand DNA replication. We will determine how the TnsE protein interacts with a target DNA and core machinery to recruit the rest of the transposition apparatus. The transposition pathway will be established in a reconstituted in vitro reaction to understand the mechanism of target site selection at the molecular level. Relevance to Public Health: We will use a highly tractable bacterial system to get a better understanding of how DNA replication and repair function. This will provide much needed insight into very similar systems in humans which when non-functional can lead to cancer. Public health will be served because a better understanding of DNA repair and replication provides insight into the genesis of cancer.

描述（由申请人提供）：实验室的长期目标是了解生物体如何在分子水平上保持其基因组信息。我们正在研究基因组在一个叫做转座的过程中被改变的方法之一。转座子是可在细胞内移动的移动的DNA元件，其几乎存在于每种生物体中并且在人类基因组中丰富。为了更好地理解分子水平上的转座，我们使用细菌转座子Tn 7。Tn 7转座特别令人感兴趣，因为它还使我们能够理解DNA复制和修复，因为Tn 7识别参与这些过程的复合物作为插入靶点。这项工作与人类健康有关，因为它将导致更好地了解所有生物体中发现的细胞DNA修复功能，这些功能对预防癌症很重要。Tn 7能够使用Tn 7编码的蛋白质TnsE识别与多种形式的DNA代谢相关的结构或复合物。TnsE介导的转座优先发生在能够在细胞之间移动的质粒中，称为接合质粒。TnSE介导的途径还识别与DNA双链断裂修复相关的结构，并允许Tn 7优先指导DNA复制终止的转座。我们将确定TnsE是否直接与β-钳持续合成因子相互作用以识别滞后链DNA复制。我们将确定TnsE蛋白如何与靶DNA和核心机制相互作用，以招募其余的转座装置。转座途径将建立在一个重建的体外反应，以了解在分子水平上的靶位点选择的机制。与公共卫生的相关性：我们将使用一个高度易处理的细菌系统来更好地了解DNA复制和修复功能。这将为人类非常相似的系统提供急需的洞察力，这些系统在无功能时可能导致癌症。公共卫生将得到服务，因为更好地了解DNA修复和复制提供了深入了解癌症的起源。