Genome-wide RNAi screening identifies human proteins with a regulatory function in the early secretory pathway

Jeremy C. Simpson,1 Brigitte Joggerst,2 Vibor Laketa,2 Fatima Verissimo,2 Cihan Cetin,2 Holger Erfle,2, 6 Mariana G. Bexiga,1 Vasanth R. Singan,1 Jean-Karim Hériché,3 Beate Neumann,3 Alvaro Mateos,2 Jonathon Blake,4 Stephanie Bechtel,5 Vladimir Benes,4 Stefan Wiemann,5 Jan Ellenberg2, 3 & Rainer Pepperkok2

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.

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