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- Title
Genome-wide RNAi screening identifies human proteins with a regulatory function in the early secretory pathway.
- Authors
Simpson, Jeremy C.; Joggerst, Brigitte; Laketa, Vibor; Verissimo, Fatima; Cetin, Cihan; Erfle, Holger; Bexiga, Mariana G.; Singan, Vasanth R.; Hériché, Jean-Karim; Neumann, Beate; Mateos, Alvaro; Blake, Jonathon; Bechtel, Stephanie; Benes, Vladimir; Wiemann, Stefan; Ellenberg, Jan; Pepperkok, Rainer
- 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.
- Subjects
MAMMALIAN cell cycle; RNA interference; CELL membranes; HUMAN cell culture; CYTOPLASM; G proteins; CLONING
- Publication
Nature Cell Biology, 2012, Vol 14, Issue 7, p764
- ISSN
1465-7392
- Publication type
Article
- DOI
10.1038/ncb2510