The Characterization of the Methanothermobacter Thermautotrophicus Origin of Replication and its Associated Proteins

嗜热甲烷嗜热杆菌复制起源及其相关蛋白的表征

• 批准号: 0237483
• 负责人: Zvi Kelman
• 金额: --
• 依托单位: University of Maryland Biotechnology Institute
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0237483&HistoricalAwards=false
• 关键词: Characterization; Methanothermobacter; Thermautotrophicus; Origin; Replication

Every proliferating cell must duplicate its genetic information accurately and only once per cell cycle. Any process that falls short of or exceeds these limits results in cell death or abnormal proliferation. The long-term goal of this research program is to understand the mechanisms governing DNA replication in eukarya, and this research project will focus specifically on the origin of replication, a specific location on the chromosome where replication initiates. Whereas DNA replication has been well characterized in bacteria, viruses and bacteriophages, many aspects of the process in eukarya, particularly in the initiation phase, are still unknown. The complex eukaryotic cell cycle entails numerous regulatory signals and factors from the first step of origin recognition to the final step of cell division, making it difficult to study individual components and events. An organism that may help to circumvent some of the complexity of eukaryotic replication is a prokaryote with eukaryotic-like replication processes,the archaeon Methanothermobacter thermautotrophicus (Mth). Archaeal DNA replication appears to be more similar to the eukaryal rather than the bacterial process. However, the archaeal DNA replication apparatus appears to require fewer proteins and complexes. Thus, archaea may provide a good model system for understanding the enzymology of eukaryotic replication. The research will focus on the mechanisms of the initiation of DNA replication in Mth.Both wild-type and mutant recombinant proteins will be used in in vitro experiments designed to:1) isolate and identify the origin binding proteins,2)understand how the origin is recognized and bound by the initiation proteins; and 3) identify the functional motifs in the archaeal origin. A long-term goal is to develop an in vitro replication system for Mth; the information gained from these biochemical studies will be directly applicable to such a system.

每个增殖的细胞都必须准确地复制其遗传信息，并且每个细胞周期只能复制一次。任何降低或超过这些限制的过程都会导致细胞死亡或异常增殖。该研究计划的长期目标是了解主持Eukarya中DNA复制的机制，该研究项目将专门关注复制的起源，复制的起源是复制启动的染色体上的特定位置。尽管DNA复制在细菌，病毒和噬菌体中都得到了很好的特征，但在Eukarya过程中，尤其是在开始阶段，该过程的许多方面仍然未知。复杂的真核细胞周期需要从原点识别的第一步到细胞分裂的最后一步，因此很难研究单个组件和事件。一种可能有助于规避真核复制的某些复杂性的生物是具有真核样复制过程的原核生物，即甲基甲基动物菌（MTH）。古细胞DNA复制似乎与真核生物而不是细菌过程更相似。但是，古细胞DNA复制设备似乎需要较少的蛋白质和复合物。因此，古细菌可以提供一个良好的模型系统，以理解真核复制的酶学。该研究将重点介绍MTH中DNA复制的机制。野生型和突变体重组蛋白将用于旨在：1）隔离和识别原点结合蛋白的体外实验，2）了解起源如何被起始蛋白识别和结合； 3）确定古细胞起源中的功能基序。一个长期目标是为MTH开发体外复制系统。从这些生化研究中获得的信息将直接适用于这种系统。