Assembly of intermediates for rapid membrane fusion.
J Biol Chem
; 293(4): 1346-1352, 2018 01 26.
Article
em En
| MEDLINE
| ID: mdl-29208657
ABSTRACT
Membrane fusion is essential for intracellular protein sorting, cell growth, hormone secretion, and neurotransmission. Rapid membrane fusion requires tethering and Sec1-Munc18 (SM) function to catalyze R-, Qa-, Qb-, and Qc-SNARE complex assembly in trans, as well as SNARE engagement by the SNARE-binding chaperone Sec17/αSNAP. The hexameric vacuolar HOPS (homotypic fusion and vacuole protein sorting) complex in the yeast Saccharomyces cerevisiae tethers membranes through its affinities for the membrane Rab GTPase Ypt7. HOPS also has specific affinities for the vacuolar SNAREs and catalyzes SNARE complex assembly, but the order of their assembly into a 4-SNARE complex is unclear. We now report defined assembly intermediates on the path to membrane fusion. We found that a prefusion intermediate will assemble with HOPS and the R, Qa, and Qc SNAREs, and that this assembly undergoes rapid fusion upon addition of Qb and Sec17. HOPS-tethered membranes and all four vacuolar SNAREs formed a complex that underwent an even more dramatic burst of fusion upon Sec17p addition. These findings provide initial insights into an ordered fusion pathway consisting of the following intermediates and events 1) Rab- and HOPS-tethered membranes, 2) a HOPSRQaQc trans-complex, 3) a HOPS4-SNARE trans-complex, 4) an engagement with Sec17, and 5) the rapid lipid rearrangements during fusion. In conclusion, our results indicate that the RQaQc complex forms in the context of membrane, Ypt7, HOPS, and trans-SNARE assembly and serves as a functional intermediate for rapid fusion after addition of the Qb-SNARE and Sec17 proteins.
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01-internacional
Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
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Membrana Celular
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Proteínas de Saccharomyces cerevisiae
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Proteínas SNARE
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Fusão de Membrana
Idioma:
En
Revista:
J Biol Chem
Ano de publicação:
2018
Tipo de documento:
Article