SM protein Sly1 and a SNARE Habc domain promote membrane fusion through multiple mechanisms.
Journal
The Journal of cell biology
ISSN: 1540-8140
Titre abrégé: J Cell Biol
Pays: United States
ID NLM: 0375356
Informations de publication
Date de publication:
03 Jun 2024
03 Jun 2024
Historique:
received:
17
01
2020
revised:
20
12
2023
accepted:
22
02
2024
pmc-release:
13
09
2024
medline:
15
3
2024
pubmed:
13
3
2024
entrez:
13
3
2024
Statut:
ppublish
Résumé
SM proteins including Sly1 are essential cofactors of SNARE-mediated membrane fusion. Using SNARE and Sly1 mutants and chemically defined in vitro assays, we separate and assess proposed mechanisms through which Sly1 augments fusion: (i) opening the closed conformation of the Qa-SNARE Sed5; (ii) close-range tethering of vesicles to target organelles, mediated by the Sly1-specific regulatory loop; and (iii) nucleation of productive trans-SNARE complexes. We show that all three mechanisms are important and operate in parallel, and that close-range tethering promotes trans-complex assembly when cis-SNARE assembly is a competing process. Further, we demonstrate that the autoinhibitory N-terminal Habc domain of Sed5 has at least two positive activities: it is needed for correct Sed5 localization, and it directly promotes Sly1-dependent fusion. "Split Sed5," with Habc presented solely as a soluble fragment, can function both in vitro and in vivo. Habc appears to facilitate events leading to lipid mixing rather than promoting opening or stability of the fusion pore.
Identifiants
pubmed: 38478017
pii: 276617
doi: 10.1083/jcb.202001034
pmc: PMC10943372
pii:
doi:
Substances chimiques
Munc18 Proteins
0
Qa-SNARE Proteins
0
Saccharomyces cerevisiae Proteins
0
SNARE Proteins
0
Vesicular Transport Proteins
0
Sed5 protein, S cerevisiae
0
SLY1 protein, S cerevisiae
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM077349
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM130644
Pays : United States
Organisme : NIH HHS
ID : R01 GM077349
Pays : United States
Informations de copyright
© 2024 Duan et al.
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