Zinc Finger Protein-Protein Interactions

转录因子之间的蛋白质-蛋白质相互作用在转录调控中起着关键作用。 正常和疾病状态下的基因表达。本申请提出研究蛋白质-蛋白质 由Cys 2 His 2锌指蛋白介导的相互作用， 人类的因素。特别是，我们正在集中研究二聚锌指（DZF） 在转录因子的Ikaros家族成员和相关同源物中发现的结构域。 DZF结构域为研究Cys 2 His 2锌指介导的蛋白质-蛋白质提供了模型系统 交互.该提案的长期目标是：获得详细的生物物理理解 Cys 2 His 2锌指如何介导与各种不同蛋白质的特异性相互作用; 利用这些知识来设计策略，以增强，破坏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， 用于创建在生物技术中具有潜在应用的人工调节电路的模块， 治疗学本申请的具体目标是：1）定义和建模Ikaros，飞马座， 和Hunchback DZF结构域接口，使用突变分析，遗传选择方法， 分子图形工具; 2）确定二聚体Ikaros，Pegasus的高分辨率结构， Hunchback DZF结构域蛋白质-蛋白质复合物使用X射线晶体学; 3）使用遗传学方法， 选择以鉴定合成的专性异二聚体DZF结构域并测试DZF结构域的用途 作为在真核细胞中组装异源蛋白质复合物的“相互作用模块”。

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.