Proteomic analysis reveals a PLK1-dependent G2/M degradation program and a role for AKAP2 in coordinating the mitotic cytoskeleton

Ubiquitination is an essential regulator of cell division. The kinase Polo-like kinase 1 (PLK1) promotes protein degradation at G2/M phase through the E3 ubiquitin ligase Skp1-Cul1-F box (SCF)βTrCP. However, the magnitude to which PLK1 shapes the mitotic proteome is uncharacterized. Combining quantitative proteomics with pharmacologic PLK1 inhibition revealed a widespread, PLK1-dependent program of protein breakdown at G2/M. We validated many PLK1-regulated proteins, including substrates of the cell-cycle E3 SCFCyclin F, demonstrating that PLK1 promotes proteolysis through at least two distinct E3 ligases. We show that the protein-kinase-A-anchoring protein A-kinase anchor protein 2 (AKAP2) is cell-cycle regulated and that its mitotic degradation is dependent on the PLK1/βTrCP signaling axis. Expression of a non-degradable AKAP2 mutant resulted in actin defects and aberrant mitotic spindles, suggesting that AKAP2 degradation coordinates cytoskeletal organization during mitosis. These findings uncover PLK1’s far-reaching role in shaping the mitotic proteome post-translationally and have potential implications in malignancies where PLK1 is upregulated. [Display omitted] •PLK1 promotes a widespread program of protein degradation at G2/M phase of the cell cycle•PLK1-mediated degradation is coordinated through at least two SCF-family E3 ligases•AKAP2, a PKA-anchoring protein, is regulated by the PLK1/βTrCP signaling axis•AKAP2 degradation coordinates cytoskeletal dynamics in mitosis Mouery et al. utilize deep, quantitative proteomics to reveal a PLK1-regulated program of G2/M protein degradation. They show that the PKA-anchoring protein AKAP2 is degraded in a manner dependent on PLK1 and the E3 ubiquitin ligase SCFβTrCP and that disruption of AKAP2 degradation results in defects of the mitotic cytoskeleton.