Comprehensive Protein Interaction Mapping in Mammals

描述(由申请人提供)： 了解特定蛋白质之间的相互作用可以为它们的功能提供重要的联系。对模式生物、酵母和线虫在基因组范围内发生的相互作用的理解正在进行中。来自哺乳动物系统的类似信息是非常可取的。然而，哺乳动物基因组的复杂性大约增加了10倍，导致评估所有潜在相互作用的难度呈指数级增加。本提案将制定方法，以提高已成功地在模式生物中建立全基因组蛋白质相互作用图的技术可应用于哺乳动物系统的效率。这里要修改的具体技术是传统的酵母双杂交系统。一旦建立了这些方法，它们将足够强大，可以使用单个高通量毛细管测序仪在不到3个月的时间内为哺乳动物细胞中可能发生的所有1×10(10)潜在相互作用建立完整的蛋白质相互作用图。基于相似蛋白质相互作用图在模式生物中的应用，这项工作有望为哺乳动物细胞的功能提供巨大的见解。

DESCRIPTION (provided by applicant): Knowledge of the interactions occurring between specific proteins can provide important links to their functions. Attempts to develop an understanding of the interactions that occur on a genome wide scale for model organisms, yeast and nematodes, are ongoing. Similar information form mammalian systems is highly desirable. However, the approximately 10 fold greater complexity of the mammalian genome leads to an exponential increase in the difficulty of assessing all of the potential interactions. The present proposal will develop methodologies that enhance the efficiency with which techniques that have been successful in establishing genome wide protein interaction maps in model organisms can be applied to mammalian systems. The specific technique that will be modified here is the traditional yeast two-hybrid system. Once the methodologies are established, they will be sufficiently powerful to allow a complete protein interaction map to be established for all of the 1 x 10(10) potential interactions that are possible in a mammalian cell within less than 3 months using a single high throughput capillary sequencer. Based on the utility of similar protein interaction maps in model organisms, this work can be expected to provide an enormous wealth of insight into the function of mammalian cells.