Comprehensive Protein Interaction Mapping in Mammals

描述（由申请人提供）： 了解特定蛋白质之间发生的相互作用可以为它们的功能提供重要的联系。目前正在尝试了解模式生物酵母和线虫在全基因组范围内发生的相互作用。从哺乳动物系统中获得类似的信息是非常可取的。然而，哺乳动物基因组的复杂性增加了约10倍，导致评估所有潜在相互作用的难度呈指数级增加。本提案将开发提高效率的方法，已成功地建立全基因组蛋白质相互作用图的模式生物的技术可以应用于哺乳动物系统。这里要改进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存在的所有1 × 10（10）种潜在相互作用建立完整的蛋白质相互作用图谱。基于类似的蛋白质相互作用图在模式生物中的效用，这项工作可以预期提供了大量的洞察哺乳动物细胞的功能。

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.