1. Genome-wide RNAi screening identifies human proteins with a regulatory function in the early secretory pathway
- Author
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Jean-Karim Hériché, Mariana G. Bexiga, Stefan Wiemann, Alvaro Mateos, Jonathon Blake, Holger Erfle, Jan Ellenberg, Rainer Pepperkok, Vladimir Benes, Brigitte Joggerst, Jeremy C. Simpson, Vibor Laketa, Stephanie Bechtel, Beate Neumann, Vasanth R. Singan, Fatima Verissimo, and Cihan Cetin
- Subjects
Signal peptide ,Time Factors ,Recombinant Fusion Proteins ,Golgi Apparatus ,Biology ,Transfection ,Actin cytoskeleton organization ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,Humans ,Secretion ,Gene Regulatory Networks ,Cloning, Molecular ,Transport Vesicles ,COPII ,Secretory pathway ,030304 developmental biology ,Monomeric GTP-Binding Proteins ,Platelet-Derived Growth Factor ,0303 health sciences ,Epidermal Growth Factor ,Secretory Vesicles ,Cell Biology ,COPI ,Golgi apparatus ,Endocytosis ,Cell biology ,Actin Cytoskeleton ,Protein Transport ,Secretory protein ,Gene Expression Regulation ,Microscopy, Fluorescence ,symbols ,RNA Interference ,030217 neurology & neurosurgery ,HeLa Cells ,Signal Transduction - 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.
- Published
- 2012