Examining the Role of Exo V in Repairing DNA Damage

描述（由申请人提供）：生命取决于遗传信息从一代到下一代的忠实传递。不幸的是，DNA损伤会发生，当它发生时，可能会导致重要遗传信息的丢失。我的工作重点是了解两组不同的细菌如何选择解决一种非常有害的DNA损伤类型所带来的问题：双链断裂。本提案的第一部分集中在RecBCD，这是一种来自E.杆菌RecBCD是一种有效的双链5 '-3'和3 '-5'核酸酶、解旋酶和ATP酶。它是高度进行性的，每次结合事件消化DNA的酶。然而，它也是促进同源重组的重组酶。这两个矛盾的作用是由8个核苷酸的chi序列（5 '-GCTGGTGG-3'）介导的。该提案的第二部分集中在AddAB上，AddAB是T.腾冲菌等革兰氏阳性菌。AddAB在细胞中具有与RecBCD相同的活性和作用，除了它响应于不同的chi序列。在这个项目中，我将使用各种技术，包括晶体学，遗传学和酶学，以确定结构和解剖这些加工核酸酶的调节。

DESCRIPTION (provided by applicant): Life depends upon the faithful transmission of genetic information from one generation to the next. Unfortunately DNA damage occurs and, when it does, it can lead to the loss of important genetic information. My work focuses on understanding how two different groups of bacteria have chosen to solve the problem presented by one very harmful type of DNA damage: double-strand breaks. The first part of this proposal focuses on RecBCD, which is a heterotrimeric protein from E. coli. RecBCD is a potent double strand 5'-3' and 3'-5' nuclease, helicase, and ATPase. It is highly processive, digesting kilobases of DNA per binding event. It is, however, also a recombinase that promotes homologous recombination. These two paradoxical roles are mediated by the 8-nucleotide chi sequence (5'-GCTGGTGG-3'). The second part of the proposal focuses on AddAB, the two-subunit enzyme responsible for Exo V activity in T. tengcongensis and other gram-positive bacteria. AddAB has the same activity and role in the cell as RecBCD except it responds to a different chi sequence. In this project I will use a variety of techniques including crystallography, genetics, and enzymology to determine the structure and dissect the regulation of these processive nucleases.