Phosphoproteomic Analysis of Aurora Kinase Inhibition in Monopolar Cytokinesis

Cytokinesis
is the last step of the cell cycle that requires coordinated
activities of the microtubule cytoskeleton, actin cytoskeleton, and
membrane compartments. Aurora B kinase is one of the master regulatory
kinases that orchestrate multiple events during cytokinesis. To reveal
targets of the Aurora B kinase, we combined quantitative mass spectrometry
with chemical genetics. Using the quantitative proteomic approach,
SILAC (stable isotope labeling with amino acids in cell culture),
we analyzed the phosphoproteome of monopolar cytokinesis upon VX680-
or AZD1152-mediated aurora kinase inhibition. In total, our analysis
quantified over 20 000 phosphopeptides in response to the Aurora-B
kinase inhibition; 246 unique phosphopeptides were significantly down-regulated
and 74 were up-regulated. Our data provide a broad analysis of downstream
effectors of Aurora kinase and offer insights into how Aurora kinase
regulates cytokinesis.