Coupling between clathrin-dependent endocytic budding and F-BAR-dependent tubulation in a cell-free
Author: ["Min Wu","Bo Huang","Morven Graham","Andrea Raimondi","John E. Heuser","Xiaowei Zhuang","Pietro De Camilli"]
Publication: Nature Cell Biology
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Abstract
A cell-free system has been developed to image vesicle budding and fission events. This method reveals an important role for the F-BAR protein FBP17 in regulating tubulation and clathrin-dependent budding. Cell-free reconstitution of membrane traffic reactions and the morphological characterization of membrane intermediates that accumulate under these conditions have helped to elucidate the physical and molecular mechanisms involved in membrane transport1,2,3. To gain a better understanding of endocytosis, we have reconstituted vesicle budding and fission from isolated plasma membrane sheets and imaged these events. Electron and fluorescence microscopy, including subdiffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)4,5,6, revealed F-BAR (FBP17) domain coated tubules nucleated by clathrin-coated buds when fission was blocked by GTPγS. Triggering fission by replacing GTPγS with GTP led not only to separation of clathrin-coated buds, but also to vesicle formation by fragmentation of the tubules. These results suggest a functional link between FBP17-dependent membrane tubulation and clathrin-dependent budding. They also show that clathrin spatially directs plasma membrane invaginations that lead to the generation of endocytic vesicles larger than those enclosed by the coat.
Cite this article
Wu, M., Huang, B., Graham, M. et al. Coupling between clathrin-dependent endocytic budding and F-BAR-dependent tubulation in a cell-free system. Nat Cell Biol 12, 902–908 (2010). https://doi.org/10.1038/ncb2094
