Sequential SNARE disassembly and GATE-16-GOS-28 complex assembly mediated by distinct NSF activities drives Golgi membrane fusion.
Characterization of mammalian NSF (G274E) and Drosophila NSF (comatose) mutants revealed an evolutionarily conserved NSF activity distinct from ATPase-dependent SNARE disassembly that was essential for Golgi membrane fusion. Analysis of mammalian NSF function during cell-free assembly of Golgi cisternae from mitotic Golgi fragments revealed that NSF disassembles Golgi SNAREs during mitotic Golgi fragmentation. A subsequent ATPase-independent NSF activity restricted to the reassembly phase is essential for membrane fusion. NSF/alpha-SNAP catalyze the binding of GATE-16 to GOS-28, a Golgi v-SNARE, in a manner that requires ATP but not ATP hydrolysis. GATE-16 is essential for NSF-driven Golgi reassembly and precludes GOS-28 from binding to its cognate t-SNARE, syntaxin-5. We suggest that this occurs at the inception of Golgi reassembly to protect the v-SNARE and regulate SNARE function.
Muller
Joyce M M
JM
Endothelial Cell Biology, Cancer Research UK, London WC2A 3PX, United Kingdom.
Shorter
James
J
Newman
Richard
R
Deinhardt
Katrin
K
Sagiv
Yuval
Y
Elazar
Zvulun
Z
Warren
Graham
G
Shima
David T
DT
D000251:Adenosine Triphosphatases / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D001693:Biological Transport, Active; D016466:CHO Cells; D002352:Carrier Proteins / Q000502:physiology*; D002478:Cells, Cultured; D006224:Cricetinae; D004330:Drosophila; D019143:Evolution, Molecular; D006056:Golgi Apparatus / Q000378:metabolism / Q000502:physiology*; D007425:Intracellular Membranes / Q000502:physiology*; D008561:Membrane Fusion / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis; D009154:Mutation; D052066:N-Ethylmaleimide-Sensitive Proteins; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins; C498732:comt protein, Drosophila; D000251:Adenosine Triphosphatases; D052066:N-Ethylmaleimide-Sensitive Proteins