Roles of the TRAPP-II Complex and the Exocyst in Membrane Deposition during Fission Yeast Cytokinesis

Abstract

The cleavage-furrow tip adjacent to the actomyosin contractile ring is believed to be the predominant site for plasma-membrane insertion through exocyst-tethered vesicles during cytokinesis. Here we found that most secretory vesicles are delivered by myosin-V on linear actin cables in fission yeast cytokinesis. Surprisingly, by tracking individual exocytic and endocytic events, we found that vesicles with new membrane are deposited to the cleavage furrow relatively evenly during contractile-ring constriction, but the rim of the cleavage furrow is the main site for endocytosis. Fusion of vesicles with the plasma membrane requires vesicle tethers. Our data suggest that the transport particle protein II (TRAPP-II) complex and Rab11 GTPase Ypt3 help to tether secretory vesicles or tubulovesicular structures along the cleavage furrow while the exocyst tethers vesicles at the rim of the division plane. We conclude that the exocyst and TRAPP-II complex have distinct localizations at the division site, but both are important for membrane expansion and exocytosis during cytokinesis.

Abstract

Two putative vesicle tethers—the exocyst and TRAPP-II complexes—localize differently at the division plane to ensure efficient plasma-membrane deposition along the whole cleavage furrow during cytokinesis in the fission yeast Schizosaccharomyces pombe.

Author Summary

Cytokinesis partitions a mother cell into two daughter cells at the end of each cell-division cycle. A significant amount of new plasma membrane is needed at the cleavage furrow during cytokinesis in many cell types. Membrane expansion is achieved through the balance of exocytosis and endocytosis. It is poorly understood where and when the membrane is deposited and retrieved during cytokinesis. By tracking individual vesicles with high spatiotemporal resolution and using electron microscopy, we found that new membrane is deposited relatively evenly along the cleavage furrow in fission yeast, while the rim of the division plane is the predominant site for endocytosis. The secretory vesicles/compartments carrying new membrane are mainly delivered along formin-nucleated actin cables by myosin-V motors. Surprisingly, we find that both exocytosis and endocytosis at the division site are ramped up before contractile-ring constriction and last until daughter-cell separation. We discovered that two putative vesicle tethers, the exocyst and TRAPP-II complexes, localize to different sites at the cleavage furrow to promote tethering of different, yet overlapping, classes of secretory vesicles/compartments for exocytosis and new membrane deposition.