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| Title | Structural basis for activation of Arf1 at the Golgi complex. |
|---|---|
| Journal, issue, pages | Cell Rep, Vol. 40, Issue 9, Page 111282, Year 2022 |
| Publish date | Aug 30, 2022 |
 Authors | Arnold J Muccini / Margaret A Gustafson / J Christopher Fromme /  |
| PubMed Abstract | The Golgi complex is the central sorting station of the eukaryotic secretory pathway. Traffic through the Golgi requires activation of Arf guanosine triphosphatases that orchestrate cargo sorting and ...The Golgi complex is the central sorting station of the eukaryotic secretory pathway. Traffic through the Golgi requires activation of Arf guanosine triphosphatases that orchestrate cargo sorting and vesicle formation by recruiting an array of effector proteins. Arf activation and Golgi membrane association is controlled by large guanine nucleotide exchange factors (GEFs) possessing multiple conserved regulatory domains. Here we present cryoelectron microscopy (cryoEM) structures of full-length Gea2, the yeast paralog of the human Arf-GEF GBF1, that reveal the organization of these regulatory domains and explain how Gea2 binds to the Golgi membrane surface. We find that the GEF domain adopts two different conformations compatible with different stages of the Arf activation reaction. The structure of a Gea2-Arf1 activation intermediate suggests that the movement of the GEF domain primes Arf1 for membrane insertion upon guanosine triphosphate binding. We propose that conformational switching of Gea2 during the nucleotide exchange reaction promotes membrane insertion of Arf1. |
 External links | Cell Rep / PubMed:36044848 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.8 - 4.7 Å |
| Structure data | EMDB-26716, PDB-7uro: EMDB-26717, PDB-7urr:  EMDB-26749: Consensus map of Gea2-Arf1 activation intermediate complex  EMDB-26750: Focused map of DCB-HUS domains of the Gea2-Arf1 activation intermediate complex  EMDB-26751: Focused map of GEF domain-Arf1 in the Gea2-Arf1 activation intermediate complex  EMDB-26752: Focused map of HDS1,2,3 domains in the Gea2-Arf1 activation intermediate complex  EMDB-26753: Focused map of the dimer interface of the Gea2-Arf1 activation intermediate complex EMDB-26754, PDB-7ut4:  EMDB-26755: Consensus map of Gea2 in the closed conformation  EMDB-26765: Focused map of the N-terminal region of Gea2 in the closed conformation  EMDB-26766: Focused map of the GEF domain of Gea2 in the closed conformation  EMDB-26769: Focused map of the C-terminal region of Gea2 in the closed conformation EMDB-26770, PDB-7uth:  EMDB-26771: Consensus map of Gea2 in open conformation  EMDB-26773: Focused map of the N-terminal region of Gea2 in the open conformation  EMDB-26774: Focused map of the GEF domain of Gea2 in the open conformation  EMDB-26775: Focused map of the C-terminal region of Gea2 in the closed conformation  EMDB-26776: Focused map of the dimer interface of Gea2 in the open conformation  EMDB-26777: Focused map of the dimer interface of Gea2 in the closed conformation  EMDB-26778: Focused map of the dimer interface of Gea2 in closed/open conformation  EMDB-26779: Focused map of the closed state N-terminal region of Gea2 closed/open conformation  EMDB-26780: Focused map of the closed state GEF domain of Gea2 closed/open conformation  EMDB-26781: Focused map of the closed C-terminal region of Gea2 closed/open conformation  EMDB-26783: Focused map of the open state N-terminal region of Gea2 closed/open conformation  EMDB-26784: Focused map of the open state GEF domain of Gea2 closed/open conformation  EMDB-26785: Focused map of the open state C-terminal region Gea2 closed/open conformation  EMDB-26797: Consensus map of Gea2 closed/open conformation |
| Source |
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 Keywords | PROTEIN TRANSPORT / GEF |
saccharomyces cerevisiae (brewer's yeast)