title: RNAi screening reveals a large signaling network controlling the Golgi apparatus in human cells.
abstract: 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.
authors.affiliation: Laboratory Regulation of Membrane Traffic, Institute of Molecular and Cell Biology, Singapore, Singapore.
authors.forename: Joanne
authors.initials: J
authors.lastname: Chia
authors.forename: Germaine
authors.initials: G
authors.lastname: Goh
authors.forename: Victor
authors.initials: V
authors.lastname: Racine
authors.forename: Susanne
authors.initials: S
authors.lastname: Ng
authors.forename: Pankaj
authors.initials: P
authors.lastname: Kumar
authors.forename: Frederic
authors.initials: F
authors.lastname: Bard
mesh_terms: D000205:Actomyosin / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D019295:Computational Biology; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D037102:Lectins / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence; D010641:Phenotype; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D010865:Pilot Projects; D011134:Polysaccharides / Q000096:biosynthesis; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D015203:Reproducibility of Results; D015398:Signal Transduction
chemical_list: D037102:Lectins; D011134:Polysaccharides; D000205:Actomyosin; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases