Interactions between N-terminal Modules in MPS1 Enable Spindle Checkpoint Silencing

Faithful chromosome segregation relies on the ability of the spindle assembly checkpoint (SAC) to delay anaphase onset until chromosomes are attached to the mitotic spindle via their kinetochores. MPS1 kinase is recruited to kinetochores to initiate SAC signaling and is removed from kinetochores once stable microtubule attachments have been formed to allow normal mitotic progression. Here, we show that a helical fragment within the kinetochore-targeting N-terminal extension (NTE) module of MPS1 is required for interactions with kinetochores and forms intramolecular interactions with its adjacent tetratricopeptide repeat (TPR) domain. Bypassing this NTE-TPR interaction results in high MPS1 levels at kinetochores due to loss of regulatory input into MPS1 localization, inefficient MPS1 delocalization upon microtubule attachment, and SAC silencing defects. These results show that SAC responsiveness to attachments relies on regulated intramolecular interactions in MPS1 and highlight the sensitivity of mitosis to perturbations in the dynamics of the MPS1-NDC80-C interactions. [Display omitted] •MPS1 kinase localizes to kinetochores via a helical fragment in its NTE module•The NTE can interact with the neighboring TPR domain•The NTE-TPR interaction regulates the binding of MPS1 to kinetochores•Disrupting this interaction perturbs SAC silencing without affecting MT attachments Pachis et al. show that proper binding dynamics of the spindle checkpoint kinase MPS1 to kinetochores are ensured by regulated intramolecular interactions. Perturbing these interactions can lead to inefficient displacement of MPS1 upon microtubule attachment and defects in inactivating the spindle assembly checkpoint, which is necessary for timely chromosome segregation.