A Drosophila geneome-wide protein interaction map

描述（由申请人提供）：相互作用的蛋白质网络介导多种生物过程。对生物系统进行更深入了解的一个重要步骤是对构成每个基因组的这些蛋白质网络的所有二元相互作用进行编目。该项目的长期目标是为遗传易学的模式生物果蝇（Drosophila melanogaster）构建一个全面、高质量的蛋白质相互作用图谱。酵母双杂交系统是检测生物信息性蛋白质相互作用的成熟技术。在这个项目完成的阶段，酵母双杂交克隆阵列构建了超过86%的预测果蝇基因14000。这些阵列的初步筛选检测到数千种蛋白质相互作用，这对于理解蛋白质和途径功能是非常宝贵的。分析还显示，到目前为止，只有一小部分可以用这种技术检测到的相互作用被确定。该项目的第一个目标是通过持续的双杂交筛选来增加蛋白质相互作用数据的覆盖率，特别是目前相互作用图中缺失的蛋白质；这将使现有的交互数量至少增加一倍。在第二个目标中，将使用两种不同的分析系统来验证所有检测到的相互作用。使用这些结果和其他标准，每个相互作用将被分配一个置信度分数，表明它在体内发挥功能作用的概率。在第三个目标中，将执行共亲和纯化以进一步验证相互作用数据。在Aim 4中，二元相互作用将直接测试怀疑是功能相关的蛋白质组或计算预测相互作用的蛋白质之间的相互作用。与随机筛选相比，这种方法可能产生更完整和准确的这些蛋白质相互作用数据，并且应该为验证和进一步增强预测算法提供数据。在最终目标中，将克隆现有双杂交阵列中缺失的新预测基因并将其纳入筛选过程。该项目的所有数据将通过至少两个独立的基于网络的数据库以标准化格式公开提供。综合起来，这个项目的数据将为理解保守蛋白网络的功能和建模复杂的生物过程和疾病提供基础。

DESCRIPTION (provided by applicant): Networks of interacting proteins mediate a diverse range of biological processes. An essential step toward developing a more thorough understanding of biological systems is to catalogue all of the binary interactions that comprise these protein networks for each genome. The long-term objective of this project is to construct a comprehensive high-quality protein interaction map for the genetically tractable model organism, Drosophila melanogaster. A proven technology for detecting biologically informative protein interactions is the yeast two-hybrid system. In a completed phase of this project, yeast two-hybrid clone arrays were constructed for over 86% of the 14,000 predicted Drosophila genes. Initial screening of these arrays detected thousands of protein interactions, which are proving invaluable for understanding protein and pathway function. Analyses have also revealed that only a small fraction of the interactions detectable with this technology have been identified thus far. The first aim of this project is to increase the coverage of protein interaction data by continued two-hybrid screening, particularly with the proteins missing from current interaction maps; this will at least double the existing number of interactions. In the second aim, two different assay systems will be used to verify all detected interactions. Using these results and other criteria, each interaction will be assigned a confidence score indicating the probability that it plays a functional role in vivo. In the third aim, co-affinity purifications will be performed to further validate the interaction data. In Aim 4, binary interactions will be directly tested for groups of proteins suspected of being functionally related or between proteins computationally predicted to interact. Compared to random screening, this approach is likely to produce more complete and accurate interaction data for these proteins, and moreover, should provide data to help validate and further enhance prediction algorithms. In the final aim, newly predicted genes missing from the existing two-hybrid arrays will be cloned and incorporated into the screening process. All data from this project will be publicly available in standardized formats via at least two independent web-based databases. Combined, the data from this project will provide a foundation for understanding the functions of conserved protein networks and for modeling complex biological processes and diseases.