Lipid packing sensed by ArfGAP1 couples COPI coat disassembly to membrane bilayer curvature.
Protein coats deform flat lipid membranes into buds and capture membrane proteins to form transport vesicles. The assembly/disassembly cycle of the COPI coat on Golgi membranes is coupled to the GTP/GDP cycle of the small G protein Arf1. At the heart of this coupling is the specific interaction of membrane-bound Arf1-GTP with coatomer, a complex of seven proteins that forms the building unit of the COPI coat. Although COPI coat disassembly requires the catalysis of GTP hydrolysis in Arf1 by a specific GTPase-activating protein (ArfGAP1), the precise timing of this reaction during COPI vesicle formation is not known. Using time-resolved assays for COPI dynamics on liposomes of controlled size, we show that the rate of ArfGAP1-catalysed GTP hydrolysis in Arf1 and the rate of COPI disassembly increase over two orders of magnitude as the curvature of the lipid bilayer increases and approaches that of a typical transport vesicle. This leads to a model for COPI dynamics in which GTP hydrolysis in Arf1 is organized temporally and spatially according to the changes in lipid packing induced by the coat.
Bigay
Joëlle
J
Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, 660 route des Lucioles, 06560 Valbonne-Sophia-Antipolis, France.
Gounon
Pierre
P
Robineau
Sylviane
S
Antonny
Bruno
B
D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D000818:Animals; D022181:COP-Coated Vesicles / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D020755:Coat Protein Complex I / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D007425:Intracellular Membranes / Q000737:chemistry; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D011817:Rabbits; D013997:Time Factors
D020755:Coat Protein Complex I; D020690:GTPase-Activating Proteins; D008051:Lipid Bilayers; D008055:Lipids; D008081:Liposomes; D006160:Guanosine Triphosphate; D020727:ADP-Ribosylation Factors