Phosphorylation controls spatial and temporal activities of motor-PRC1 complexes to complete mitosis

Abstract

During mitosis, spindle architecture alters as chromosomes segregate to daughter cells. The microtubule crosslinker Protein Required for Cytokinesis 1 (PRC1) is essential for spindle stability, chromosome segregation and completion of cytokinesis, but how it recruits motors to the central spindle to coordinate the segregation of chromosomes is unknown. Here, we combine structural and cell biology approaches to show that the human CENP-E motor, which is essential for chromosome capture and alignment by microtubules, binds to PRC1 through a conserved hydrophobic motif. This binding mechanism is also used by Kinesin-4 Kif4A:PRC1. Using in vitro reconstitution, we demonstrate that CENP-E slides antiparallel PRC1-crosslinked microtubules. We find that the regulation of CENP-E -PRC1 interaction is spatially and temporally coupled with relocalization to overlapping microtubules in anaphase. Finally, we demonstrate that the PRC1:microtubule motor interaction is essential in anaphase to control chromosome partitioning, retain central spindle integrity and ensure cytokinesis. Taken together our findings reveal the molecular basis for the cell cycle regulation of motor-PRC1 complexes to couple chromosome segregation and cytokinesis.

Kinesin-7 CENP-E in tumorigenesis: Chromosome instability, spindle assembly checkpoint, and applications

“…The combination of CENP-E and kinetochore protein Ndc80 supports lateral transport and microtubule wall-to-end transition at stabilized microtubules (Chakraborty et al, 2019). CENP-E binds to PRC1 through a conserved hydrophobic motif and promotes the antiparallel PRC1-crosslinked microtubules (Gluszek-Kustusz et al, 2023). During spindle assembly, PRC1-crosslinked microtubules undergo a network-to-bundles transition, and CENP-E promotes further microtubule bundling and kinetochore-mediated overlap formation (Matković et al, 2022).…”