Function and Structure of Bacterial RNA Ligase

描述(申请人提供)：我们实验室最近在耐辐射球菌中发现了一种新的具有功能的RNA连接酶。由于在其他几种生物中也可以发现同源蛋白质，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，本研究关注的编码葡萄球菌RNA连接酶的基因位于其他假定的RNA修复途径成员附近。在我提议的研究过程中，我将仔细检查这种新酶的底物要求，确定一个跨属保守但功能未知的新结构域的作用，识别有助于催化的残基，以及表征它们贡献的性质。除了这些体外实验，我还将检测这种RNA连接酶在体内的作用，并使用X射线结晶学来确定天然结构。

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.