Max Planck Institute for Molecular Cell Biology and Genetics, Germany.
Nature. 2010 Mar 11;464(7286):243-9. doi: 10.1038/nature08779. Epub 2010 Feb 28.
Endocytosis is a complex process fulfilling many cellular and developmental functions. Understanding how it is regulated and integrated with other cellular processes requires a comprehensive analysis of its molecular constituents and general design principles. Here, we developed a new strategy to phenotypically profile the human genome with respect to transferrin (TF) and epidermal growth factor (EGF) endocytosis by combining RNA interference, automated high-resolution confocal microscopy, quantitative multiparametric image analysis and high-performance computing. We identified several novel components of endocytic trafficking, including genes implicated in human diseases. We found that signalling pathways such as Wnt, integrin/cell adhesion, transforming growth factor (TGF)-beta and Notch regulate the endocytic system, and identified new genes involved in cargo sorting to a subset of signalling endosomes. A systems analysis by Bayesian networks further showed that the number, size, concentration of cargo and intracellular position of endosomes are not determined randomly but are subject to specific regulation, thus uncovering novel properties of the endocytic system.
内吞作用是一个复杂的过程，具有许多细胞和发育功能。要了解它是如何被调节的，以及如何与其他细胞过程相整合，就需要对其分子成分和一般设计原则进行全面分析。在这里，我们开发了一种新的策略，通过结合 RNA 干扰、自动化高分辨率共聚焦显微镜、定量多参数图像分析和高性能计算，来表型分析人类基因组与转铁蛋白（TF）和表皮生长因子（EGF）内吞作用有关的方面。我们鉴定了几个内吞运输的新成分，包括涉及人类疾病的基因。我们发现，信号通路，如 Wnt、整合素/细胞黏附、转化生长因子（TGF）-β和 Notch 调节内吞系统，并鉴定了参与货物分拣到信号内体亚群的新基因。贝叶斯网络的系统分析进一步表明，内体的数量、大小、货物浓度和细胞内位置不是随机决定的，而是受到特定的调节，从而揭示了内吞系统的新特性。
