Dynamics of eukaryotic junction-resolving enzyme GEN1 - DNA junction interactions

细胞DNA损伤的修复对人体健康至关重要，其中一个重要的修复过程就是重组。这包括在细胞DNA中形成分支结构，然后由酶处理。其中一种酶被称为GEN1。我们最近已经解决了GEN1蛋白的原子结构，这种结构为可能的作用机制提供了相当大的见解。我们现在将通过观察酶的单分子来扩展我们的知识，因为它们使用高度敏感的荧光方法进行连接的处理。我们将能够解剖蛋白质与DNA结合并在分支连接处组装的过程，以及它如何改变DNA的结构。然后，我们可以继续研究酶在特定点切割连接DNA的过程，以及这是如何与结构变化相结合的。我们的目标是充分了解这种酶如何实现良好调节的连接过程。我们的最终目标是用小分子干预这个过程，最终产生治疗剂。

Repair of damage to cellular DNA is extremely important to human health, and one important repair process is recombination. This involves the formation of branched structures in cellular DNA, that are then processed by enzymes. One such enzyme is called GEN1. We have recently solved the atomic structure of the GEN1 protein, and this structure has provided considerable insight into the probable mechanism of action. We now shall extend our knowledge by observation of single molecules of the enzyme as they carry out the processing of junctions, using highly sensitive fluorescence methods. We will be able to dissect the processes by which the protein binds to the DNA and assembles on the branched junction, and how it alters the structure of the DNA. We can then proceed to study the process by which the enzyme cuts the DNA of the junction at specific points, and how this is coupled to changes in structure. Our goal is a full understanding of how this enzyme achieves a well regulated processing of the junction. Our ultimate goal is to intervene in the process using small molecules, which could ultimately result in therapeutic agents.