Genome-wide Investigation of PDZ Domain Specificity

PDZ 结构域特异性的全基因组研究

• 批准号: 7011218
• 负责人: GAVIN MACBEATH
• 金额: $ 30.43万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7011218
• 关键词: Internet; animal genetic material tag; animal population genetics; fluorescence polarization; gel filtration chromatography; gene expression; genetic screening; genome; intermolecular interaction; laboratory mouse; ligands; microarray technology; molecular biology information system; molecular cloning; protein protein interaction; protein purification; protein structure function; transcription factor

DESCRIPTION (provided by applicant): Most eukaryotic proteins that receive and process signals are constructed from a combination of interaction and catalytic domains. Among the many interaction domains identified in the past decade, PDZ's are one of the most frequently encountered. They are often found in combination with other interaction modules and play a role in directing the specificity of receptor tyrosine kinase-mediated signaling, in establishing cell polarity, in directing protein trafficking, and in coordinating synaptic signaling. Their importance is underscored by the severe neuronal and developmental phenotypes observed in PDZ knockout mice and by their implication in human congenital diseases like Usher syndrome and Dejerine-Sottas neuropathy. The enormous diversity of PDZ function is manifest in their abundance; there are over 250 PDZ's encoded in the mouse genome. To understand their individual roles, it is necessary first to define their recognition properties in a comprehensive and relevant fashion. Previous efforts to define PDZ selectivity have focused on only a few domains and have relied on collections of randomized peptides, rather than on physiological ligands. In order to provide a genome-wide understanding of PDZ function, the selectivity of every PDZ encoded in the mouse genome will be investigated relative to a large collection of genomically-encoded ligands using protein microarray technology. The aims of this proposal are: (1) to clone, express, and purify every mouse PDZ domain; (2) to screen every PDZ with every PDZ to reveal putative PDZ-PDZ interactions; (3) to screen every PDZ with 222 genomically-encoded peptide ligands to identify putative PDZ-protein interactions; (4) to investigate the physiological relevance of a subset of predicted interactions biochemically; and (5) to construct a relational database that integrates protein microarray data with published information on PDZ domains. Collectively, these efforts should provide the necessary foundation to understand PDZ function on a genome-wide level and aid future efforts to intervene appropriately when PDZ function goes awry.

描述（由申请人提供）： 大多数接收和过程信号的真核蛋白都是由相互作用和催化域的组合构建的。在过去十年中确定的许多交互域中，PDZ是最常遇到的一个。它们通常与其他相互作用模块结合使用，并在指导受体酪氨酸激酶介导的信号传导的特异性，在建立细胞极性，指导蛋白质运输和协调突触信号传导方面发挥作用。 在PDZ敲除小鼠中观察到的严重神经元和发育表型强调了它们的重要性，以及它们对人类先天性疾病（如Usher综合征和Dejerine-Sottas Neuropathy）的影响。 PDZ功能的巨大多样性在其丰度中表现出来。小鼠基因组中编码了250多个PDZ。要了解他们的个人角色，有必要首先以全面和相关的方式定义他们的认可属性。以前定义PDZ选择性的努力仅集中在一些领域，并且依赖于随机肽的集合，而不是生理配体。 为了提供对PDZ功能的全基因组的理解，将使用蛋白质微阵列技术研究小鼠基因组中每个PDZ的选择性。该提案的目的是：（1）克隆，表达和净化每个小鼠PDZ域； （2）用每个PDZ筛选每个PDZ，以揭示假定的PDZ-PDZ相互作用； （3）用222个基因组编码的肽配体筛选每个PDZ，以鉴定推定的PDZ蛋白质相互作用； （4）研究预测相互作用的生物化学相关性的生理相关性； （5）构建一个关系数据库，该数据库将蛋白质微阵列数据与已发布的PDZ域信息集成在一起。总的来说，这些努力应为在全基因组水平上了解PDZ功能的必要基础，并帮助未来的努力在PDZ功能出现时适当干预。