Dynein inhibition (bottom) prevents removal of Mad2 (red) from kinetochores.
Previously, Howell et al. found that the Mad2 checkpoint protein is transported from kinetochores to the spindle poles along spindle microtubules. The authors have now found that the microtubule motors CENP-E and cytoplasmic dynein, as well as the checkpoint-related proteins BubR1 and the 3F3/2 antigen, are also depleted from kinetochores and accumulate at the spindle poles. Inhibiting dynein/dynactin activity stops this transport process and simultaneously blocks mitosis at metaphase.
The data suggest that proteins in the outer kinetochore domain maintain a dynamic equilibrium that shifts as the kinetochore interacts with microtubules. Once the association pathways are blocked, constitutive dissociation pathways, including dynein/dynactin transport, deplete proteins from the metaphase kinetochores and inactivate the checkpoint. Recent results in Drosophila melanogaster, in which mutations in cytoplasmic dynein cause checkpoint-related proteins to accumulate at metaphase kinetochores, underscore the importance of dynein in kinetochore disassembly (Wojcik, E., et al. 2001. Nat. Cell Biol. 3:1001–1007).▪