Careful examination of the morphological changes of the vacuole
during salt-induced fragmentation allowed us to dissect the process
into two distinct phases with nonoverlapping requirements for
the known fragmentation factors. This dissection and the fact that
vesiculation happens in an asymmetrical manner at sites that are
identifiable in the light microscope provides an important tool for
future identification of additional proteins involved in vacuole fragmentation
and for studying how they shape the membrane.