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The Journal of Cell Biology


Vacuole inheritance in yeast involves the for- mation of tubular and vesicular “segregation struc- tures” which migrate into the bud and fuse there to es- tablish the daughter cell vacuole. Vacuole fusion has been reconstituted in vitro and may be used as a model for an NSF-dependent reaction of priming, docking, and fusion. We have developed biochemical and micro- scopic assays for the docking step of in vitro vacuole fusion and characterized its requirements. The vacu- oles must be primed for docking by the action of Sec17p ( a -SNAP) and Sec18p (NSF). Priming is neces- sary for both fusion partners. It produces a labile state which requires rapid docking in order to lead produc- tively to fusion. In addition to Sec17p/Sec18p, docking requires the activity of the Ras-like GTPase Ypt7p. Un- like Sec17p/Sec18p, which must act before docking, Ypt7p is directly involved in the docking process itself.