Spatial Exclusivity Combined with Positive and Negative Selection of Phosphorylation Motifs Is the Basis for Context-Dependent Mitotic Signaling

Sci. Signal., 28 June 2011
Vol. 4, Issue 179, p. ra42
DOI: 10.1126/scisignal.2001796

Spatial Exclusivity Combined with Positive and Negative Selection of Phosphorylation Motifs Is the Basis for Context-Dependent Mitotic Signaling

  1. Jes Alexander1,*,
  2. Daniel Lim1,
  3. Brian A. Joughin1,
  4. Björn Hegemann2,,
  5. James R. A. Hutchins2,
  6. Tobias Ehrenberger1,
  7. Frank Ivins3,
  8. Fabio Sessa4,
  9. Otto Hudecz2,
  10. Erich A. Nigg5,
  11. Andrew M. Fry6,
  12. Andrea Musacchio4,
  13. P. Todd Stukenberg7,
  14. Karl Mechtler2,
  15. Jan-Michael Peters2,
  16. Stephen J. Smerdon3, and
  17. Michael B. Yaffe1,8,
  1. 1Koch Institute for Integrative Cancer Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
  2. 2Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, A-1030 Vienna, Austria.
  3. 3Division of Molecular Structure, Medical Research Council National Institute for Medical Research, The Ridgeway, London NW7 1AA, UK.
  4. 4Department of Experimental Oncology, European Institute of Oncology, Via Adamello 16, I-20139 Milan, Italy.
  5. 5Biozentrum, University of Basel, Klingelbergstrasse 50/70, CH-4056 Basel, Switzerland.
  6. 6Department of Biochemistry, University of Leicester, Leicester LE1 9HN, UK.
  7. 7Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
  8. 8Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
  1. To whom correspondence should be addressed. E-mail: myaffe{at}mit.edu
  • * Present address: Department of Radiation Oncology, University of California, San Francisco, CA 94143, USA.

  • Present address: Institute of Biochemistry, Eidgenössische Technische Hochschule Zürich, Schafmattstrasse 18, CH-8093 Zürich, Switzerland.

Abstract

The timing and localization of events during mitosis are controlled by the regulated phosphorylation of proteins by the mitotic kinases, which include Aurora A, Aurora B, Nek2 (never in mitosis kinase 2), Plk1 (Polo-like kinase 1), and the cyclin-dependent kinase complex Cdk1/cyclin B. Although mitotic kinases can have overlapping subcellular localizations, each kinase appears to phosphorylate its substrates on distinct sites. To gain insight into the relative importance of local sequence context in kinase selectivity, identify previously unknown substrates of these five mitotic kinases, and explore potential mechanisms for substrate discrimination, we determined the optimal substrate motifs of these major mitotic kinases by positional scanning oriented peptide library screening (PS-OPLS). We verified individual motifs with in vitro peptide kinetic studies and used structural modeling to rationalize the kinase-specific selection of key motif-determining residues at the molecular level. Cross comparisons among the phosphorylation site selectivity motifs of these kinases revealed an evolutionarily conserved mutual exclusion mechanism in which the positively and negatively selected portions of the phosphorylation motifs of mitotic kinases, together with their subcellular localizations, result in proper substrate targeting in a coordinated manner during mitosis.

Citation:

J. Alexander, D. Lim, B. A. Joughin, B. Hegemann, J. R. Hutchins, T. Ehrenberger, F. Ivins, F. Sessa, O. Hudecz, E. A. Nigg, A. M. Fry, A. Musacchio, P. T. Stukenberg, K. Mechtler, J.-M. Peters, S. J. Smerdon, and M. B. Yaffe, Spatial Exclusivity Combined with Positive and Negative Selection of Phosphorylation Motifs Is the Basis for Context-Dependent Mitotic Signaling. Sci. Signal. 4, ra42 (2011).

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