Muller Joyce M M, Shorter James, Newman Richard, Deinhardt Katrin, Sagiv Yuval, Elazar Zvulun, Warren Graham, Shima David T
Endothelial Cell Biology, Cancer Research UK, London WC2A 3PX, United Kingdom.
J Cell Biol. 2002 Jun 24;157(7):1161-73. doi: 10.1083/jcb.200202082. Epub 2002 Jun 17.
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.
对哺乳动物NSF（G274E）和果蝇NSF（昏迷）突变体的表征揭示了一种进化上保守的NSF活性，该活性不同于依赖ATP酶的SNARE拆解，是高尔基体膜融合所必需的。对有丝分裂高尔基体片段进行无细胞高尔基体潴泡组装过程中哺乳动物NSF功能的分析表明，NSF在有丝分裂高尔基体碎片化过程中拆解高尔基体SNARE。随后一种仅限于重新组装阶段的不依赖ATP酶的NSF活性对于膜融合至关重要。NSF/α-SNAP以一种需要ATP但不需要ATP水解的方式催化GATE-16与高尔基体v-SNARE GOS-28的结合。GATE-16对于NSF驱动的高尔基体重新组装至关重要，并阻止GOS-28与其同源t-SNARE syntaxin-5结合。我们认为这发生在高尔基体重新组装开始时，以保护v-SNARE并调节SNARE功能。
