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Investigation of a soluble DNA translocase

可溶性 DNA 易位酶的研究

基本信息

批准号：
203277908
负责人：
Professor Dr. Peter Graumann
金额：
--
依托单位：
Zentrum für Synthetische Mikrobiologie (SYNMIKRO)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2014-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/203277908?language=en
关键词：
Investigation soluble DNA translocase

项目摘要

Cells must ensure that duplicated chromosomes are fully separated from each other before cell division occurs. While it is still unclear how eukaryotic cells prevent the entrapment of segregation-delayed chromosomes within the division septum, it has become clear that bacteria contain dedicated DNA translocases that transport DNA through closed membranes if cell division occurs before segregation has been completed. In the case of the model bacterium Bacillus subtilis, we have recently discovered that two DNA translocases perform this task at two different steps. The newly identified SftA DNA translocase moves DNA away during the closure of the division septum, while the membrane protein SpoIIIE pumps DNA through a channel it forms within the closed septum. Interestingly, we found that SftA is fully soluble, and is thus the first non-membrane bound DNA translocase described so far. We plan to investigate how the protein binds to DNA, a question that has not been answered for any DNA translocase. SftA forms hexamers in solution, which somehow must open up for DNA translocation. We will also investigate how the DNA translocation motor of SftA works at a molecular level. We will also analyse how SftA is recruited to the division machinery, which itself is membrane-anchored, through the identification of direct binding partners of SftA.

在细胞分裂发生之前，细胞必须确保复制的染色体彼此完全分离。虽然目前尚不清楚真核细胞是如何防止分离延迟染色体在分裂隔膜内的包裹，但已经清楚的是，如果细胞分裂发生在分离完成之前，细菌含有专用的DNA转座，通过封闭的膜运输DNA。在模式细菌枯草芽孢杆菌的情况下，我们最近发现两个DNA转位在两个不同的步骤执行这项任务。新发现的SftA DNA转位酶在分裂隔膜关闭期间将DNA移走，而膜蛋白SpoIIIE则通过封闭隔膜内形成的通道泵送DNA。有趣的是，我们发现SftA是完全可溶的，因此是迄今为止描述的第一个非膜结合DNA转位酶。我们计划研究这种蛋白质是如何与DNA结合的，这是任何DNA转位酶都无法回答的问题。SftA在溶液中形成六聚体，它必须以某种方式为DNA易位打开。我们还将研究SftA的DNA易位马达在分子水平上是如何工作的。我们还将通过鉴定SftA的直接结合伙伴，分析SftA如何被招募到分裂机制中，而分裂机制本身是膜锚定的。