Assembly and Dynamics of the DNA Replication Machine

DNA 复制机的组装和动力学

• 批准号: 435521-2013
• 负责人: ReyesLamothe, Rodrigo
• 金额: $ 3.5万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=629838
• 关键词: Assembly; Dynamics; DNA; Replication; Machine

Cells in our body carry the same genetic information as that of the embryo we once were. Coded in DNA, the information is reliably transferred from mother to daughter cells thousands of times as we grow. Molecular machines work inside the cells to copy and maintain this genetic information. The study of these machines, and others working in different cellular processes, has been hindered by a major limitation of biochemistry: the impossibility of reconstructing intracellular conditions in test tubes.Outlined in this proposal is an alternative strategy to study DNA replication, one that involves moving away from attempts to reconstruct intracellular condition in vitro and instead obtaining relevant biochemical information directly from living cells. The long-term objective of this laboratory is to extract detailed mechanistic insights on the processes of DNA metabolism directly from living cells by capitalizing on recent advances in the field of microscopy. While in the current proposal the candidate will investigate two specific aspects of the DNA replication machine: its assembly early during the cell cycle and the mechanisms that prevent its premature disassembly prior to the completion of DNA replication.This work will help furthering a novel approach for the study of molecular events happening within cells. Use of this approach to study the assembly and stability of the replication machinery will provide unprecedented insight towards their understanding. Increased comprehension on the DNA replication machine in bacteria will help understanding how similar machines copy and maintain the chromosome in all other organisms.

我们体内的细胞携带着与我们曾经的胚胎相同的遗传信息。这些信息被编码在DNA中，随着我们的成长，这些信息会可靠地从母体细胞传给子代细胞数千次。分子机器在细胞内工作，复制和维护这些遗传信息。对这些机器以及其他在不同细胞过程中工作的机器的研究，一直受到生物化学的一个主要限制：不可能在试管中重建细胞内条件。这项建议概述了研究DNA复制的另一种策略，即放弃在体外重建细胞内条件的尝试，而是直接从活细胞中获得相关的生化信息。这个实验室的长期目标是通过利用显微镜领域的最新进展，直接从活细胞中提取关于DNA新陈代谢过程的详细机制见解。而在目前的提案中，候选人将研究DNA复制机器的两个具体方面：它在细胞周期的早期组装，以及在DNA复制完成之前防止其过早分解的机制。这项工作将有助于进一步发展一种新的方法来研究细胞内发生的分子事件。使用这种方法来研究复制机械的组装和稳定性将为他们的理解提供前所未有的洞察力。增加对细菌DNA复制机器的理解将有助于理解类似的机器是如何复制和维护所有其他生物的染色体的。