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TitleSubtomogram averaging of COPII assemblies reveals how coat organization dictates membrane shape.
Journal, issue, pagesNat Commun, Vol. 9, Issue 1, Page 4154, Year 2018
Publish dateOct 8, 2018
AuthorsJoshua Hutchings / Viktoriya Stancheva / Elizabeth A Miller / Giulia Zanetti /
PubMed AbstractEukaryotic cells employ membrane-bound carriers to transport cargo between compartments in a process essential to cell functionality. Carriers are generated by coat complexes that couple cargo ...Eukaryotic cells employ membrane-bound carriers to transport cargo between compartments in a process essential to cell functionality. Carriers are generated by coat complexes that couple cargo capture to membrane deformation. The COPII coat mediates export from the endoplasmic reticulum by assembling in inner and outer layers, yielding carriers of variable shape and size that allow secretion of thousands of diverse cargo. Despite detailed understanding of COPII subunits, the molecular mechanisms of coat assembly and membrane deformation are unclear. Here we present a 4.9 Å cryo-tomography subtomogram averaging structure of in vitro-reconstituted membrane-bound inner coat. We show that the outer coat (Sec13-Sec31) bridges inner coat subunits (Sar1-Sec23-Sec24), promoting their assembly into a tight lattice. We directly visualize the membrane-embedded Sar1 amphipathic helix, revealing that lattice formation induces parallel helix insertions, yielding tubular curvature. We propose that regulators like the procollagen receptor TANGO1 modulate this mechanism to determine vesicle shape and size.
External linksNat Commun / PubMed:30297805 / PubMed Central
MethodsEM (subtomogram averaging) / EM (tomography)
Resolution4.9 Å
Structure data

0044

6gni

EMDB-0044: Cryo-tomography and subtomogram averaging of Sar1-Sec23-Sec24.
PDB-6gni: Cryo-tomography and subtomogram averaging of Sar1-Sec23-Sec24 - fitted model.
Method: EM (subtomogram averaging) / Resolution: 4.9 Å

EMDB-0307:
Representative tomogram of the COPII in vitro reconstitution used for subtomogram averaging
Method: EM (tomography)

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-GNP:
PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER / GppNHp, GMPPNP, energy-carrying molecule analogue*YM / 5'-Guanylyl imidodiphosphate

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • Saccharomyces cerevisiae (brewer's yeast)
KeywordsPROTEIN TRANSPORT / COPII coat / membrane trafficking

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