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， 独特的功能。 该酶已被证明与 转录机制并参与定义起源 DNA复制。 RNase H最近被认为与 额外的DNA校对功能。 三维结构 和未配体的RNA酶H的骨架动力学已经被表征， 然而，没有详细的结构或动力学信息， RNA酶H与底物或抑制剂复合。 因此，具体 RNase H和核酸之间的相互作用是不可用的， RNase H和螺旋DNA/RNA底物的构象变化 尚未被确认。 该研究将使用一个 组合方法来鉴定对RNase H特异的抑制剂。 在 除了组合方法之外，定点诱变将 以产生稳定的底物/RNase H复合物。 三个- RNase H复合物的空间结构和动力学性质 将使用3D和4D三重共振NMR光谱 技术. 这项研究将确定动态和结构 参与蛋白质-核酸识别和酶促的决定簇 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.