Function and Structure of Bacterial RNA Ligase

描述（由申请人提供）：我们的实验室最近在耐辐射异常球菌中鉴定了一种新型的功能性RNA连接酶。由于同源蛋白质可以在其他几种生物中找到，我们预计这项研究的发现将转化为更广泛的生物现象。此外，本研究关注的编码异常球菌RNA连接酶的基因位于其他假定的RNA修复途径成员附近。在我提出的研究过程中，我将仔细研究这种新酶的底物要求，确定一个新的结构域的作用，这是跨属保守，但未知的功能，并确定有助于催化的残基，以及表征其贡献的性质。除了这些体外实验，我还将研究这种RNA连接酶在体内的作用，并使用X射线晶体学来确定天然结构。

DESCRIPTION (provided by applicant): Our lab has recently identified a novel and functional RNA ligase in the bacterium Deinococcus radiodurans. Because homologous proteins can be found in several other organisms, we anticipate the finding of this research to translate to a broader biological phenomenon. Additionally, the gene encoding the Deinococcus RNA ligase on which this study focuses is located adjacent to other putative RNA repair pathway members. In the course of my proposed research I will carefull yexamine the substrate requirements for this new enzyme, determine the role of a novel domain that is conserved across genera but of unknown function, and identify residues that contribute to catalysis, as well as characterize the nature of their contributions. In addition to these in vitro experiments, I will also examine the role of this RNA ligase in vivo and use X-ray crystallography to determine the native structure.