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Structure paper

Title9Å structure of the COPI coat reveals that the Arf1 GTPase occupies two contrasting molecular environments.
Journal, issue, pagesElife, Vol. 6, Year 2017
Publish dateJun 16, 2017
AuthorsSvetlana O Dodonova / Patrick Aderhold / Juergen Kopp / Iva Ganeva / Simone Röhling / Wim J H Hagen / Irmgard Sinning / Felix Wieland / John A G Briggs /
PubMed AbstractCOPI 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 ...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.
External linksElife / PubMed:28621666 / PubMed Central
MethodsEM (subtomogram averaging) / X-ray diffraction
Resolution2.57 - 17.3 Å
Structure data

3720

5nzr

EMDB-3720, PDB-5nzr:
The structure of the COPI coat leaf
Method: EM (subtomogram averaging) / Resolution: 9.2 Å

3721

5nzs

EMDB-3721, PDB-5nzs:
The structure of the COPI coat leaf in complex with the ArfGAP2 uncoating factor
Method: EM (subtomogram averaging) / Resolution: 10.1 Å

3722

5nzt

EMDB-3722, PDB-5nzt:
The structure of the COPI coat linkage I
Method: EM (subtomogram averaging) / Resolution: 17.0 Å

3723

5nzu

EMDB-3723, PDB-5nzu:
The structure of the COPI coat linkage II
Method: EM (subtomogram averaging) / Resolution: 15.0 Å

3724

5nzv

EMDB-3724, PDB-5nzv:
The structure of the COPI coat linkage IV
Method: EM (subtomogram averaging) / Resolution: 17.3 Å

PDB-5mu7:
Crystal Structure of the beta/delta-COPI Core Complex
Method: X-RAY DIFFRACTION / Resolution: 2.57 Å

Chemicals

ChemComp-HOH:
WATER / Water

Source
  • mus musculus (house mouse)
  • saccharomyces cerevisiae (brewer's yeast)
  • rattus norvegicus (Norway rat)
  • chaetomium thermophilum var. thermophilum dsm 1495 (fungus)
KeywordsPROTEIN TRANSPORT / coatomer / COPI / beta COP / delta COP / TRANSPORT PROTEIN / coated vesicles / ArfGAP2

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