Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from chromosome biorientation

Abstract

Listen

Translate article

Accurate chromosome segregation depends on biorientation, whereby sister chromatids attach to microtubules emanating from opposite spindle poles. The spindle assembly checkpoint is a conserved surveillance mechanism in eukaryotes that inhibits anaphase onset until all chromosomes are bioriented1, 2, 3. In current models, the recruitment of Mad2, via Mad1, to improperly attached kinetochores is a key step needed to stop cell cycle progression3, 4, 5, 6. However, it is not known if the localization of Mad1-Mad2 to kinetochores is sufficient to block anaphase. Furthermore, it is unclear if other signalling proteins (e.g. Aurora kinases7) that regulate chromosome biorientation have checkpoint functions downstream of Mad1-Mad2 recruitment to kinetochores or if they act upstream to merely quench the primary error signal8. Here, to address both these issues, we engineered a Mad1 construct which, unlike endogenous Mad1, localizes to kinetochores that are bioriented. We show that Mad1’s constitutive localization at kinetochores is sufficient for a metaphase arrest that depends on Mad1-Mad2 binding. By uncoupling the checkpoint from its primary error signal, we show that Aurora kinase, Mps1 and BubR1, but not Polo-like kinase, are needed to maintain the checkpoint arrest even when Mad1 is present on bi-oriented kinetochores. Together, our data suggest a model in which the biorientation errors, which recruit Mad1-Mad2 to kinetochores, may be signalled not only through Mad2’s templated activation dynamics, but also through the activity of widely-conserved kinases, to ensure the fidelity of cell division.