Popis: |
Macropinocytosis is a conserved endocytic process where cells take up medium into micron-sized vesicles. InDictyostelium, macropinocytic cups form around domains of PIP3 in the plasma membrane and extend by actin polymerization. Using lattice light-sheet microscopy, we describe how cups originate, are supported by an F-actin scaffold and shaped by a ring of actin polymerization, created around PIP3 domains. How cups close is unknown. We find two ways: lip closure, where actin polymerization at the lip is re-directed inwards; and basal closure, where it stretches the cup, eventually causing membrane delamination and vesicle sealing. Cups grow as expanding waves of actin polymerization that travel across the cell surface, capturing new membrane. We propose that cups close when these waves stall. This ‘stalled wave’ hypothesis is tested through a conceptual model, where the interplay of forces from actin polymerization and membrane tension recreates many of our observations. |