A human genome-wide screen for regulators of clathrin-coated vesicle formation reveals an unexpected role for the V-ATPase View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-01

AUTHORS

Patrycja Kozik, Nicola A. Hodson, Daniela A. Sahlender, Nikol Simecek, Christina Soromani, Jiahua Wu, Lucy M. Collinson, Margaret S. Robinson

ABSTRACT

Clathrin-mediated endocytosis is essential for a wide range of cellular functions. We used a multi-step siRNA-based screening strategy to identify regulators of the first step in clathrin-mediated endocytosis, formation of clathrin-coated vesicles (CCVs) at the plasma membrane. A primary genome-wide screen identified 334 hits that caused accumulation of CCV cargo on the cell surface. A secondary screen identified 92 hits that inhibited cargo uptake and/or altered the morphology of clathrin-coated structures. The hits include components of four functional complexes: coat proteins, V-ATPase subunits, spliceosome-associated proteins and acetyltransferase subunits. Electron microscopy revealed that V-ATPase depletion caused the cell to form aberrant non-constricted clathrin-coated structures at the plasma membrane. The V-ATPase-knockdown phenotype was rescued by addition of exogenous cholesterol, indicating that the knockdown blocks clathrin-mediated endocytosis by preventing cholesterol from recycling from endosomes back to the plasma membrane. More... »

PAGES

50

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/ncb2652

DOI

http://dx.doi.org/10.1038/ncb2652

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1045128221

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23263279


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