Zinc Finger Protein-Protein Interactions

描述(由申请人提供)：转录因子之间的蛋白质-蛋白质相互作用在正常和疾病状态下基因表达的调节中起着关键作用。这项应用建议研究由Cys2His2锌指蛋白介导的蛋白质-蛋白质相互作用，Cys2His2锌指蛋白是人类中含量最丰富的一类转录因子。特别是，我们的研究重点是在Ikaros转录因子家族成员及其相关同系物中发现的二聚化锌指(DZF)结构域。DZF结构域为研究Cys2His2锌指介导的蛋白质-蛋白质相互作用提供了一个模型系统。这项建议的长期目标是：详细了解Cys2His2锌指如何调节与各种不同蛋白质的特定相互作用；利用这些知识设计策略来增强、干扰或预测这种相互作用；以及设计具有新的相互作用特异性的合成锌指，作为有用的模块来创建人工调控电路，在生物技术和治疗学中具有潜在的应用。本应用程序的具体目标是：1)使用突变分析、遗传选择方法和分子图形学工具定义并模拟Ikaros、Pegasus和Hunchback DZF结构域的界面；2)使用X射线结晶学确定Ikaros、Pegasus和Hunchback DZF结构域的高分辨率结构；3)使用遗传选择来确定合成的专有异源二聚体DZF结构域，并测试DZF结构域作为“相互作用模块”在真核细胞中组装异源蛋白质复合体的作用。

DESCRIPTION (provided by applicant): Protein-protein interactions among transcription factors play critical roles in the regulation of gene expression in normal and disease states. This application proposes to study protein-protein interactions mediated by Cys2His2 zinc finger proteins, the most abundant class of transcription factors in humans. In particular, we are focusing our studies on dimerization zinc finger (DZF) domains found in members of the Ikaros family of transcription factors and related homologues. DZF domains provide a model system for studying Cys2His2 zinc finger-mediated protein-protein interactions. The long-term goals of this proposal are: to gain a detailed biophysical understanding of how Cys2His2 zinc fingers mediate specific interactions with a wide variety of different proteins; to utilize this knowledge to devise strategies to enhance, disrupt, or predict such interactions; and to engineer synthetic zinc fingers with novel interaction specificities that could serve as useful modules for creating artificial regulatory circuits with potential applications in biotechnology and therapeutics. The specific aims of this application are: 1) to define and model the Ikaros, Pegasus, and Hunchback DZF domain interfaces using mutational analysis, genetic selection methods, and molecular graphics tools; 2) to determine high-resolution structures of dimeric Ikaros, Pegasus, and Hunchback DZF domain protein-protein complexes using X-ray crystallography; 3) to use genetic selection to identify synthetic, obligate heterodimeric DZF domains and to test the use of DZF domains as "interaction modules" for assembling heterologous protein complexes in eukaryotic cells.