Background: To function in diverse cellular processes, the dynamic behavior of microtubules (MTs) must be differentially regulated within the cell. In budding yeast, the spindle position checkpoint (SPOC) inhibits mitotic exit in response to mispositioned spindles. To maintain SPOC-mediated anaphase arrest, astral MTs must maintain persistent interactions with and/or extend through the bud neck. However, the molecular mechanisms that ensure the stability of these interactions are not known.
Results: The presence of an MT extending through and/or interacting with the bud neck is maintained by spatial control of catastrophe and rescue, which extends MT lifetime >25-fold and controls the length of dynamic MTs within the bud compartment. Moreover, the single kinesin-8 motor Kip3 alternately mediates both catastrophe and rescue of the bud MT. Kip3 accumulates in a length-dependent manner along the lattice of MTs within the bud, yet induces catastrophe spatially near the bud tip. Rather, this accumulation of Kip3 facilitates its association with depolymerizing MT plus ends, where Kip3 promotes rescue before MTs exit the bud. MT rescue within the bud requires the tail domain of Kip3, whereas the motor domain mediates catastrophe at the bud tip. In vitro, Kip3 exerts both stabilizing and destabilizing effects on reconstituted yeast MTs.
Conclusions: The kinesin-8 Kip3 is a multifunctional regulator that differentially stabilizes and destabilizes specific MTs. Control over MT catastrophe and rescue by Kip3 defines the length and lifetime of MTs within the bud compartment of cells with mispositioned spindles. This subcellular regulation of MT dynamics is critical to maintaining mitotic arrest in response to mispositioned spindles.
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