Quantitative phosphoproteomics identifies substrates and functional modules of Aurora and Polo-like kinase activities in mitotic cells

Sci Signal. 2011 Jun 28;4(179):rs5. doi: 10.1126/scisignal.2001497.

Abstract

Mitosis is a process involving a complex series of events that require careful coordination. Protein phosphorylation by a small number of kinases, in particular Aurora A, Aurora B, the cyclin-dependent kinase-cyclin complex Cdk1/cyclinB, and Polo-like kinase 1 (Plk1), orchestrates almost every step of cell division, from entry into mitosis to cytokinesis. To discover more about the functions of Aurora A, Aurora B, and kinases of the Plk family, we mapped mitotic phosphorylation sites to these kinases through the combined use of quantitative phosphoproteomics and selective targeting of kinase activities by small-molecule inhibitors. Using this integrated approach, we connected 778 phosphorylation sites on 562 proteins with these enzymes in cells arrested in mitosis. By connecting the kinases to protein complexes, we associated these kinases with functional modules. In addition to predicting previously unknown functions, this work establishes additional substrate-recognition motifs for these kinases and provides an analytical template for further use in dissecting kinase signaling events in other areas of cellular signaling and systems biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Aurora Kinase B
  • Aurora Kinases
  • CDC2 Protein Kinase / metabolism
  • Cell Cycle Proteins / metabolism*
  • Cyclin B / metabolism
  • HeLa Cells
  • Humans
  • Mitosis / physiology*
  • Phosphorylation / physiology
  • Protein-Serine-Threonine Kinases / metabolism*
  • Proteome / metabolism*
  • Proteomics
  • Proto-Oncogene Proteins / metabolism*
  • Signal Transduction / physiology*

Substances

  • Cell Cycle Proteins
  • Cyclin B
  • Proteome
  • Proto-Oncogene Proteins
  • AURKB protein, human
  • Aurora Kinase B
  • Aurora Kinases
  • Protein-Serine-Threonine Kinases
  • polo-like kinase 1
  • CDC2 Protein Kinase