The RAS GTPase RIT1 compromises mitotic fidelity through spindle assembly checkpoint suppression

The spindle assembly checkpoint (SAC) functions as a sensor of unattached kinetochores that delays mitotic progression into anaphase until proper chromosome segregation is guaranteed.1,2 Disruptions to this safety mechanism lead to genomic instability and aneuploidy, which serve as the genetic cause of embryonic demise, congenital birth defects, intellectual disability, and cancer.3,4 However, despite the understanding of the fundamental mechanisms that control the SAC, it remains unknown how signaling pathways directly interact with and regulate the mitotic checkpoint activity. In response to extracellular stimuli, a diverse network of signaling pathways involved in cell growth, survival, and differentiation are activated, and this process is prominently regulated by the Ras family of small guanosine triphosphatases (GTPases).5 Here we show that RIT1, a Ras-related GTPase that regulates cell survival and stress response,6 is essential for timely progression through mitosis and proper chromosome segregation. RIT1 dissociates from the plasma membrane (PM) during mitosis and interacts directly with SAC proteins MAD2 and p31comet in a process that is regulated by cyclin-dependent kinase 1 (CDK1) activity. Furthermore, pathogenic levels of RIT1 silence the SAC and accelerate transit through mitosis by sequestering MAD2 from the mitotic checkpoint complex (MCC). Moreover, SAC suppression by pathogenic RIT1 promotes chromosome segregation errors and aneuploidy. Our results highlight a unique function of RIT1 compared to other Ras GTPases and elucidate a direct link between a signaling pathway and the SAC through a novel regulatory mechanism. [Display omitted] •RIT1 directly interacts with SAC core components MAD2 and p31comet•CDK1 phosphorylates RIT1 during mitosis and inhibits its interaction with the SAC•RIT1 is essential for timely progression through mitosis•Pathogenic levels of RIT1 promote chromosome segregation errors Cuevas-Navarro et al. characterize a novel function of the Ras GTPase RIT1 as a negative regulator of the spindle assembly checkpoint (SAC) through its direct interaction with MAD2 and p31comet. Moreover, disease-associated RIT1 mutations that evade protein turnover promote chromosome segregation errors and aneuploidy.