Author: ["Jeremy C. Simpson","Brigitte Joggerst","Vibor Laketa","Fatima Verissimo","Cihan Cetin","Holger Erfle","Mariana G. Bexiga","Vasanth R. Singan","Jean-Karim Hériché","Beate Neumann","Alvaro Mateos","Jonathon Blake","Stephanie Bechtel","Vladimir Benes","Stefan Wiemann","Jan Ellenberg","Rainer Pepperkok"]
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Abstract
The secretory pathway in mammalian cells has evolved to facilitate the transfer of cargo molecules to internal and cell surface membranes. Use of automated microscopy-based genome-wide RNA interference screens in cultured human cells allowed us to identify 554 proteins influencing secretion. Cloning, fluorescent-tagging and subcellular localization analysis of 179 of these proteins revealed that more than two-thirds localize to either the cytoplasm or membranes of the secretory and endocytic pathways. The depletion of 143 of them resulted in perturbations in the organization of the COPII and/or COPI vesicular coat complexes of the early secretory pathway, or the morphology of the Golgi complex. Network analyses revealed a so far unappreciated link between early secretory pathway function, small GTP-binding protein regulation, actin cytoskeleton organization and EGF-receptor-mediated signalling. This work provides an important resource for an integrative understanding of global cellular organization and regulation of the secretory pathway in mammalian cells. Pepperkok, Simpson and colleagues performed genome-wide RNAi screens in human cells to uncover regulators of the secretory pathway. They also identify protein networks with previously unappreciated roles in secretory pathway regulation.Cite this article
Simpson, J., Joggerst, B., Laketa, V. et al. Genome-wide RNAi screening identifies human proteins with a regulatory function in the early secretory pathway. Nat Cell Biol 14, 764–774 (2012). https://doi.org/10.1038/ncb2510 