Cryptococcus neoformans employs both PP1 and PP2A phosphatases to silence the spindle checkpoint

The spindle checkpoint preserves genomic integrity by delaying anaphase onset until all kinetochores achieve bi-polar microtubule attachment. While checkpoint activation is well-characterised across eukaryotes, the mechanisms governing its silencing remain less defined and are more variable in different systems. In this study, we reveal that the fungal pathogen Cryptococcus neoformans employs a dual phosphatase-mediated mechanism to silence the spindle checkpoint. We show that PP1 is recruited to kinetochores by Spc105KNL1, via conserved SILK and RVSF motifs, while PP2A-B56 is recruited to kinetochores through direct Bub1 binding via an LxxIxE motif. Disruption of either recruitment pathway leads to prolonged checkpoint signalling and mitotic defects, underscoring the critical roles of these phosphatases in checkpoint silencing and mitotic exit. These findings further establish C. neoformans as a powerful model for studying mitotic regulation in the basidiomycete lineage and identify phosphatase recruitment interfaces as potential targets for antifungal intervention.