Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Cell. 2014 Jun 5;157(6):1473-1487. doi: 10.1016/j.cell.2014.04.029.
Endocytosis is critical for cellular physiology and thus is highly regulated. To identify regulatory interactions controlling the endocytic membrane system, we conducted 13 RNAi screens on multiple endocytic activities and their downstream organelles. Combined with image analysis of thousands of single cells per perturbation and their cell-to-cell variability, this created a high-quality and cross-comparable quantitative data set. Unbiased analysis revealed emergent properties of the endocytic membrane system and how its complexity evolved and distinct programs of regulatory control that coregulate specific subsets of endocytic uptake routes and organelle abundances. We show that these subset effects allow the mapping of functional regulatory interactions and their interaction motifs between kinases, membrane-trafficking machinery, and the cytoskeleton at a large scale, some of which we further characterize. Our work presents a powerful approach to identify regulatory interactions in complex cellular systems from parallel single-gene or double-gene perturbation screens in human cells and yeast.
内吞作用对于细胞生理学至关重要，因此受到高度调控。为了确定控制内吞膜系统的调节相互作用，我们对多种内吞作用及其下游细胞器进行了 13 次 RNAi 筛选。结合对每个扰动的数千个单细胞的图像分析及其细胞间变异性，这创建了一个高质量且可交叉比较的定量数据集。无偏分析揭示了内吞膜系统的涌现特性，以及其复杂性如何演变，以及不同的调节控制程序，这些程序共同调节特定子集的内吞摄取途径和细胞器丰度。我们表明，这些子集效应允许在大规模上对激酶、膜运输机制和细胞骨架之间的功能调节相互作用及其相互作用模式进行映射，其中一些我们进一步进行了表征。我们的工作提出了一种从人类细胞和酵母中的平行单基因或双基因扰动筛选中识别复杂细胞系统中的调节相互作用的强大方法。
