9Å Structure of the COPI Coat Reveals That the Arf1 GTPase Occupies Two Contrasting Molecular Environments

Overview

Journal Elife

Specialty Biology

Date 2017 Jun 17

PMID 28621666

Citations 53

Authors

Svetlana O Dodonova

Patrick Aderhold

Juergen Kopp

Iva Ganeva

Simone Rohling

Wim J H Hagen

Irmgard Sinning

Felix Wieland

John A G Briggs

Affiliations

Soon will be listed here.

Abstract

COPI coated vesicles mediate trafficking within the Golgi apparatus and between the Golgi and the endoplasmic reticulum. Assembly of a COPI coated vesicle is initiated by the small GTPase Arf1 that recruits the coatomer complex to the membrane, triggering polymerization and budding. The vesicle uncoats before fusion with a target membrane. Coat components are structurally conserved between COPI and clathrin/adaptor proteins. Using cryo-electron tomography and subtomogram averaging, we determined the structure of the COPI coat assembled on membranes in vitro at 9 Å resolution. We also obtained a 2.57 Å resolution crystal structure of βδ-COP. By combining these structures we built a molecular model of the coat. We additionally determined the coat structure in the presence of ArfGAP proteins that regulate coat dissociation. We found that Arf1 occupies contrasting molecular environments within the coat, leading us to hypothesize that some Arf1 molecules may regulate vesicle assembly while others regulate coat disassembly.

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