STRUCTURE AND DYNAMICS OF AN RNASE H/LIGAND COMPLEX

核糖核酸酶H(RNase H)是一种参与DNA复制的酶， 独一无二的功能。这种酶已被证明与 转录机制，并参与定义起源 DNA复制。RNaseH最近被牵连到有一种 额外的DNA校对功能。立体结构 和未连接核糖核酸酶H的骨架动力学已被表征， 然而，目前还没有详细的结构或动力学信息 核糖核酸酶H与底物或抑制剂络合。因此，特定的 核糖核酸酶H与核酸之间的相互作用不可用， 核糖核酸酶H和螺旋DNA/RNA底物的构象变化 还没有确定身份。拟议的研究将使用一种 用组合方法鉴定核糖核酸酶H。 除了组合方法外，定点突变还将 得到稳定的底物/核糖核酸酶H复合体。三个人- 核糖核酸酶H络合物的空间结构和动力学性质 将使用3D和4D三重共振核磁共振波谱进行表征 技巧。这项研究将确定动力和结构 参与蛋白质-核酸识别和酶促反应的决定因素 催化作用。

Ribonuclease H (RNase H) is an enzyme involved in DNA replication with unique functionality. The enzyme has been shown to be associated with the transcription machinery and to be involved in defining the origin of DNA replication. RNase H has recently been implicated in having an additional DNA proofreading function. The three-dimensional structure and backbone dynamics for unliganded RNase H have been characterized, yet no detailed structural or dynamical information is available for RNase H complexed with substrate or inhibitor. Thus, specific interactions between RNase H and nucleic acids are unavailable and conformational changes in both RNase H and helical DNA/RNA substrate have not been identified. The proposed research will use a combinatorial approach to identify inhibitors specific for RNase H. In addition to the combinatorial approach, site directed mutagenesis will be performed to generate stable substrate/RNase H complexes. The three- dimensional structure and dynamical properties of the RNase H complexes will be characterized using 3D and 4D triple resonance NMR spectroscopic techniques. This research will identify dynamic and structural determinants involved in protein-nucleic acid recognition and enzymatic catalysis.