Nichols B J, Ungermann C, Pelham H R, Wickner W T, Haas A
Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK.
Nature. 1997 May 8;387(6629):199-202. doi: 10.1038/387199a0.
Membrane fusion is necessary both in the eukaryotic secretory pathway and for the inheritance of organelles during the cell cycle. In the secretory pathway, heterotypic fusion takes place between small transport vesicles and organelles. It requires N-ethylmaleimide-sensitive fusion protein (NSF/Sec18p), soluble NSF attachment proteins (SNAPs/Sec17p) and SNAP receptors (SNAREs). SNAREs are integral membrane proteins (v-SNAREs on vesicles, t-SNAREs on the target organelles) and are thought to provide specificity to the fusion process. It has been suggested that Sec17p and Sec18p bind to v-SNARE/t-SNARE complexes and mediate the membrane fusion event. Homotypic fusion of yeast vacuoles also requires Sec17p and Sec18p (ref. 6), but in vitro they are needed only to 'prime' the vacuoles, not for subsequent docking or fusion. It has been unclear whether these reactions involve SNAREs that are similar to those previously identified in heterotypic fusion systems and, hence, whether the actions of Sec18p/NSF and Sec17p/alpha SNAP in these systems can be compared. Here we identify typical v- and t-SNAREs on the yeast vacuolar membrane. Although both are normally present, vacuoles containing only the v-SNARE can fuse with those containing only the t-SNARE. Vacuoles containing neither SNARE cannot fuse with those containing both, demonstrating that docking is mediated by cognate SNAREs on the two organelle membranes. Even when t- and v-SNAREs are on separate membranes, Sec17p and Sec18p act at the priming stage. Their action is not required at the point of assembly of the SNARE complex, nor for the fusion event itself.
膜融合在真核生物分泌途径以及细胞周期中细胞器的遗传过程中都是必需的。在分泌途径中,小运输囊泡与细胞器之间发生异型融合。这需要N - 乙基马来酰亚胺敏感融合蛋白(NSF / Sec18p)、可溶性NSF附着蛋白(SNAPs / Sec17p)和SNAP受体(SNAREs)。SNAREs是整合膜蛋白(囊泡上的v - SNAREs,靶细胞器上的t - SNAREs),被认为为融合过程提供特异性。有人提出Sec17p和Sec18p与v - SNARE / t - SNARE复合物结合并介导膜融合事件。酵母液泡的同型融合也需要Sec17p和Sec18p(参考文献6),但在体外它们仅用于“启动”液泡,而不是用于后续的对接或融合。目前尚不清楚这些反应是否涉及与先前在异型融合系统中鉴定的那些类似的SNAREs,因此,也不清楚Sec18p / NSF和Sec17p /αSNAP在这些系统中的作用是否可以进行比较。在这里,我们鉴定了酵母液泡膜上典型的v - 和t - SNAREs。虽然两者通常都存在,但仅含有v - SNARE的液泡可以与仅含有t - SNARE的液泡融合。既不含有SNARE的液泡不能与同时含有两者的液泡融合,这表明对接是由两个细胞器膜上的同源SNAREs介导的。即使t - 和v - SNAREs位于不同的膜上,Sec17p和Sec18p也在启动阶段起作用。在SNARE复合物组装点以及融合事件本身都不需要它们的作用。