Mapping the Human Binary Interactome Network

描述（由申请人提供）：生物体的相互作用组是由一系列生理相关浓度的完整物理相互作用形成的网络，包括蛋白质-蛋白质、dna -蛋白质和rna -蛋白质相互作用。生成具有高特异性和高灵敏度的蛋白质组相互作用组网络图是在整个细胞范围内生成预测性大分子模型的必要方面，尽管不是充分方面。此外，这些图谱可作为机制和定量研究的基础，以研究特征不佳的预测基因产物和疾病相关蛋白。有两种主要的方法可以通过实验绘制任何感兴趣的生物体的蛋白质-蛋白质相互作用组网络：i)可以通过实验测试预测的蛋白质对的所有成对组合，以获得所有可能的二元物理相互作用（二元相互作用组）；ii)所有蛋白质都可以通过测试来询问它们属于哪个蛋白质复合物（共复合物相互作用组）。二元和共复方法在提供越来越精确和完整的相互作用组模型方面是互补的，这是公认的。我们建议继续努力系统地绘制人类二元相互作用组，目标是生成和分析具有高质量和高灵敏度的新版本的HT-Y2H相互作用，用于我们至少有一个网关克隆ORF可用的所有预测基因产物的成对组合。这对应于一个约16000 X 16000个蛋白质的矩阵，这代表了整个矩阵的约50%，因为我们假设总共约22000个蛋白质编码基因，并将分析限制在每个基因的单个剪接变体。

DESCRIPTION (provided by applicant): The interactome of an organism is the network formed by the complete set of physical interactions that can occur in a range of physiologically relevant concentrations, including protein-protein, DNA-protein, and RNA-protein interactions. The generation of proteome-wide interactome network maps with high specificity and high sensitivity is a necessary, although not sufficient, aspect in the quest of generating predictive macromolecular models at the scale of the whole cell. Moreover, these maps serve as a foundation for mechanistic and quantitative studies of poorly characterized predicted gene products and disease-associated proteins. There are two major approaches to experimentally map the protein-protein interactome network of any organism of interest: i) all pairwise combinations of pairs of predicted proteins can be experimentally tested to derive all possible binary physical interactions (binary interactome); and ii) all proteins can be tested to interrogate in which protein complex(es) they belong (co-complex interactome). It is well established that binary and co-complex approaches are complementary in providing increasingly accurate and complete interactome models. We propose to continue our efforts at systematically mapping the human binary interactome with the goal of generating and analyzing a new version of HT-Y2H interactions with high quality and high sensitivity for all pairwise combinations of predicted gene products for which we have at least one Gateway-cloned ORF available. This corresponds to a matrix of ~16,000 X 16,000 proteins, which represents ~50% of the complete matrix since we assume a total of ~22,000 protein-coding genes and limit the analysis to a single splice variant per gene. Our modified specific aims are to: i) Provide an expanded high-confidence/high-coverage version of the human binary interactome map ii) Validate human binary interaction data iii) Analyze the expanded human interactome model