TRIP13 Functions in the Establishment of the Spindle Assembly Checkpoint by Replenishing O-MAD2

Abstract

The spindle assembly checkpoint (SAC) prevents premature segregation of chromosomes during mitosis. This process requires structural remodeling of MAD2 from O-MAD2 to C-MAD2 conformation. After the checkpoint is satisfied, C-MAD2 is reverted to O-MAD2 to allow anaphase-promoting complex/cyclosome (APC/C) to trigger anaphase. Recently, the AAA + -ATPase TRIP13 was shown to act in concert with p31 comet to catalyze C- to O-MAD2. Paradoxically, although C-MAD2 is present in TRIP13-deficient cells, the SAC cannot be activated. Using a degron-mediated system to uncouple TRIP13 from O- and C-MAD2 equilibrium, we demonstrated that the loss of TRIP13 did not immediately abolish the SAC, but the resulting C-MAD2-only environment was insufficient to enable the SAC. These results favor a model in which MAD2-CDC20 interaction is coupled directly to the conversion of O- to C-MAD2 instead of one that involves unliganded C-MAD2. TRIP13 replenishes the O-MAD2 pool for activation by unattached kinetochores. C-MAD2 is required for the activation of the spindle assembly checkpoint. Ma and Poon show that C-MAD2 alone is insufficient to support the spindle assembly checkpoint. TRIP13 is involved in replenishing the O-MAD2 pool for conversion into C-MAD2 and the activation of the spindle assembly checkpoint.