Choreography of the assembly of proteins in DNA double strand break repair centres in bacteria

细菌 DNA 双链断裂修复中心蛋白质组装的编排

• 批准号: 37966473
• 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: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/37966473?language=en
• 关键词: Choreography; assembly; proteins; DNA; double

Many proteins involved in the repair of double strand breaks (DSBs) in DNA have been identified, and their biochemical properties have been studied to various degrees. However, the exact pathway of DSB repair is still unclear. We have recently been able to prove the existence of repair centres (RCs) that are transiently induced in response to DSBs in Bacillus subtilis cells, and to visualize the temporal and ordered recruitment of several proteins to RCs. These experiments have already provided important novel insight into DSB repair, and are an excellent system to dissect the function of DSB repair proteins in live cells. This proposal deals with repair proteins RecJ, RecX, YhaN/O, SbcC/D, RadC and Smf, which perform important functions, but in very different ways. The RecJ exonuclease functions at an early time point during DSB repair, but, like RadC, also at the replication fork in non-stressed cells. RecX has been shown to interact with the major repair protein RecA, and may modulate RecA activity. YhaN and SbcC are SMC-like proteins whose function is yet unclear. We plan to dissect the function of the genes genetically, by combining the deletions with all other known deletions in DSB genes, by protein purification and biochemical analysis, and by visualization of GFP fusions, to put the proteins into the correct context of appearance and assembly during repair.

许多参与DNA双链断裂（DSBs）修复的蛋白质已经被鉴定出来，它们的生化特性也得到了不同程度的研究。然而，DSB修复的确切途径尚不清楚。我们最近已经能够证明枯草芽孢杆菌细胞中瞬时诱导的修复中心（RCs）的存在，并可视化几种蛋白质向RCs的时间和有序募集。这些实验已经为DSB修复提供了重要的新见解，并且是一个很好的系统来解剖DSB修复蛋白在活细胞中的功能。本研究涉及修复蛋白RecJ、RecX、YhaN/O、SbcC/D、RadC和Smf，它们发挥着重要的功能，但方式却截然不同。RecJ外切酶在DSB修复的早期时间点起作用，但与RadC一样，在非应激细胞中也在复制叉上起作用。RecX已被证明与主要修复蛋白RecA相互作用，并可能调节RecA的活性。YhaN和SbcC是smc样蛋白，其功能尚不清楚。我们计划通过将缺失与DSB基因中所有其他已知缺失结合起来，通过蛋白质纯化和生化分析，以及GFP融合的可视化，从遗传学上剖析这些基因的功能，将蛋白质置于修复过程中外观和组装的正确背景中。