Data from: RNAi screening reveals a large signaling network controlling the Golgi apparatus in human cells

The Golgi apparatus has many important physiological functions, including sorting of secretory cargo and biosynthesis of complex glycans. These functions depend on the intricate and compartmentalized organization of the Golgi apparatus. To investigate the mechanisms that regulate Golgi architecture, we developed a quantitative morphological assay using three different Golgi compartment markers and quantitative image analysis, and performed a kinome- and phosphatome-wide RNAi screen in HeLa cells. Depletion of 159 signaling genes, nearly 20% of genes assayed, induced strong and varied perturbations in Golgi morphology. Using bioinformatics data, a large regulatory network could be constructed. Specific subnetworks are involved in phosphoinositides regulation, acto-myosin dynamics and mitogen activated protein kinase signaling. Most gene depletion also affected Golgi functions, in particular glycan biosynthesis, suggesting that signaling cascades can control glycosylation directly at the Golgi level. Our results provide a genetic overview of the signaling pathways that control the Golgi apparatus in human cells.

高尔基体（Golgi apparatus）具备诸多重要生理功能，包括分泌货物分选与复杂聚糖生物合成。这些功能的发挥依赖于高尔基体精密且分区化的组织结构。为探究调控高尔基体结构的分子机制，本研究构建了一套定量形态学检测方法：采用三种不同的高尔基体区室标记物结合定量图像分析，并在海拉（HeLa）细胞中开展了全激酶组（kinome）与全磷酸酶组（phosphatome）范围的RNA干扰（RNAi）筛选实验。在被检测的基因中，有159个信号通路相关基因（占比近20%）的敲低会对高尔基体形态造成显著且多样的扰动。借助生物信息学数据，我们可构建出一个大型调控网络，其中特定的子网参与了磷酸肌醇调控、肌动蛋白-肌球蛋白动力学以及丝裂原活化蛋白激酶信号通路。多数基因敲低还会影响高尔基体的功能，尤其是聚糖生物合成过程，这表明信号级联反应可直接在高尔基体层面调控糖基化。本研究结果为人类细胞中调控高尔基体的信号通路提供了一份遗传学层面的全景概述。