STRUCTURE-FUNCTION STUDIES OF DNA BINDING PROTEINS AND ENZYMES

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 本文描述的研究旨在更深入地了解几类 DNA 结合蛋白和酶的结构-功能关系。 目前，实验室的五个项目已生成需要同步加速器光束时间的晶体：（1）II型限制性内切酶间接读出的作用，（2）二价阳离子依赖性核酸酶切割DNA的立体化学切割机制，（3）四聚体II型限制性内切酶特异性的调节，（4）DNA修复蛋白对受损DNA的识别，（5）人工C2H2锌指的识别码。 许多涉及蛋白质和酶如何实现非常高的 DNA 序列辨别力的研究。 酶促机制研究模型系统中 DNA 切割的机制，该机制被认为与许多医学相关的人类核酸内切酶有很多共同点。 DNA 修复蛋白在细胞功能中非常重要，可以识别损伤以启动修复过程，并且对于细胞命运（可以是修复或程序性细胞死亡）的决策也很重要。 锌指项目是与加州大学戴维斯分校的 David Segal 教授和斯克里普斯研究所的 Carlos Barbas 教授的实验室合作的，该实验室正在努力开发新的疗法来识别和治疗癌症和艾滋病等疾病。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The studies described herein aim to gain a deeper understanding of structure-function relationships in several classes of DNA binding proteins and enzymes. Currently, five projects in the lab have generated crystals requiring synchrotron beamtime: (1) the role of indirect readout by type II restriction endonucleases, (2) the stereochemical cleavage mechanism of DNA cleavage by divalent cation dependent nucleases, (3) the modulation of specificity in a tetrameric type II restriction endonuclease, (4) the recognition of damaged DNA by DNA repair proteins, (5) the recognition code of artificial C2H2 zinc fingers. Many involve the investigation of how proteins and enzymes achieve very high DNA sequence discrimination. The enzymatic mechanisms study the mechanism of DNA cleavage in a model system, which is believed to have much in common with many medically relevant human endonucleases. The DNA repair proteins are important in cellular function in identifying damage for the initiation of the repair process, and also for decision making regarding the cell fate, which can be either repair or programmed cell death. The zinc finger project is in collaboration with the laboratories of Prof. David Segal, UC Davis, and Prof. Carlos Barbas, Scripps Institute, where efforts are under way to create new therapeutics for the identification and treatment of diseases such as cancer and AIDS.