Zinc Finger Protein-Protein Interactions

锌指蛋白-蛋白质相互作用

• 批准号: 7471666
• 负责人: J. KEITH JOUNG
• 金额: $ 1.2万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7471666
• 关键词: Affinity; Amino Acid Sequence; Binding; Biological Models; Biotechnology; Chemicals; Class; Complex; DNA; DNA Binding; DNA Sequence; Development; Dimerization; Disease; Engineering; Eukaryotic Cell; Family; Family member; Gene Expression; Gene Expression Regulation; Goals; Hematopoietic System; Homo; Homologous Gene; Human; Knowledge; Mammals; Mediating; Methods; Modeling; Molecular; Organism; Play; Process; Proteins; Range; Resolution; Role; Specificity; Structure; Testing; Therapeutic; X-Ray Crystallography; Yeasts; Zinc Fingers; design; genetic analysis; genetic selection; human disease; improved; insight; member; novel; promoter; protein protein interaction; tool; transcription factor

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.

转录因子之间的蛋白质 - 蛋白质相互作用在调节中起关键作用 正常和疾病状态中的基因表达。该应用建议研究蛋白质蛋白质 由Cys2His2锌指蛋白介导的相互作用，最丰富的转录类别 人类的因素。特别是，我们将研究重点放在二聚化锌指（DZF）上 在Ikaros家族的转录因子和相关同源物的成员中发现的域。 DZF结构域提供了一个模型系统，用于研究CYS2HIS2锌指介导的蛋白质蛋白 互动。该建议的长期目标是：获得详细的生物物理理解 cys2his2锌指的方式如何介导特定的相互作用与多种不同蛋白质的特定相互作用；到 利用这些知识来制定策略来增强，破坏或预测此类相互作用；然后 工程师合成锌指具有新型相互作用特异性的合成锌手指，可以用作有用 用于创建具有生物技术和潜在应用的人工监管电路的模块 疗法。该应用程序的具体目的是：1）定义和建模Ikaros，Pegasus， 使用突变分析，遗传选择方法和Hunchback DZF域界面 分子图形工具； 2）确定二聚体Ikaros，Pegasus和 驼背DZF域蛋白质蛋白质复合物使用X射线晶体学； 3）使用遗传 选择以识别合成，强性异二聚体DZF域并测试DZF域的使用 作为用于组装真核细胞中异源蛋白复合物的“相互作用模块”。