Charloteaux Benoit, Zhong Quan, Dreze Matija, Cusick Michael E, Hill David E, Vidal Marc
Department of Cancer Biology, Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, MA, USA.
Methods Mol Biol. 2011;759:197-213. doi: 10.1007/978-1-61779-173-4_12.
Phenotypic variations of an organism may arise from alterations of cellular networks, ranging from the complete loss of a gene product to the specific perturbation of a single molecular interaction. In interactome networks that are modeled as nodes (macromolecules) connected by edges (interactions), these alterations can be thought of as node removal and edge-specific or "edgetic" perturbations, respectively. Here we present two complementary strategies, forward and reverse edgetics, to investigate the phenotypic outcomes of edgetic perturbations of binary protein-protein interaction networks. Both approaches are based on the yeast two-hybrid system (Y2H). The first allows the determination of the interaction profile of proteins encoded by alleles with known phenotypes to identify edgetic alleles. The second is used to directly isolate edgetic alleles for subsequent in vivo characterization.
生物体的表型变异可能源于细胞网络的改变，范围从基因产物的完全缺失到单个分子相互作用的特定扰动。在被建模为通过边（相互作用）连接的节点（大分子）的相互作用组网络中，这些改变可分别被视为节点去除和边特异性或“边能学”扰动。在这里，我们提出了两种互补策略，正向和反向边能学，以研究二元蛋白质-蛋白质相互作用网络的边能学扰动的表型结果。这两种方法都基于酵母双杂交系统（Y2H）。第一种方法允许确定具有已知表型的等位基因编码的蛋白质的相互作用谱，以识别边能学等位基因。第二种方法用于直接分离边能学等位基因，以便随后进行体内表征。
