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

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...

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Main Authors: Simpson, Jeremy (Author) , Joggerst, Brigitte (Author) , Laketa, Vibor (Author) , Erfle, Holger (Author) , Bechtel, Stephanie (Author) , Wiemann, Stefan (Author) , Ellenberg, Jan (Author) , Pepperkok, Rainer (Author)
Format: Article (Journal)
Language:English
Published: 3 June 2012
In: Nature cell biology
Year: 2012, Volume: 14, Issue: 7, Pages: 764-774
ISSN:1476-4679
DOI:10.1038/ncb2510
Online Access:Verlag, Volltext: http://dx.doi.org/10.1038/ncb2510
Verlag, Volltext: https://www.nature.com/articles/ncb2510
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Author Notes: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 and Rainer Pepperkok
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Summary: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.
Item Description:Gesehen am 18.09.2018
Physical Description:Online Resource
ISSN:1476-4679
DOI:10.1038/ncb2510

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