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Science 299 (5610): 1228-1231

Copyright © 2003 by the American Association for the Advancement of Science

Proteomic Screen Finds pSer/pThr-Binding Domain Localizing Plk1 to Mitotic Substrates

Andrew E. H. Elia,12 Lewis C. Cantley,2 Michael B. Yaffe1*

We have developed a proteomic approach for identifying phosphopeptide binding domains that modulate kinase-dependent signaling pathways. An immobilized library of partially degenerate phosphopeptides biased toward a particular protein kinase phosphorylation motif is used to isolate phospho-binding domains that bind to proteins phosphorylated by that kinase. Applying this approach to cyclin-dependent kinases (Cdks), we identified the polo-box domain (PBD) of the mitotic kinase polo-like kinase 1 (Plk1) as a specific phosphoserine (pSer) or phosphothreonine (pThr) binding domain and determined its optimal binding motif. This motif is present in known Plk1 substrates such as Cdc25, and an optimal phosphopeptide containing the motif disrupted PBD-substrate binding and localization of the PBD to centrosomes. This finding reveals how Plk1 can localize to specific sites within cells in response to Cdk phosphorylation at those sites and provides a structural mechanism for targeting the Plk1 kinase domain to its substrates.

1 Center for Cancer Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
2 Division of Signal Transduction, Beth Israel Deaconess Hospital, and Department of Cell Biology, Harvard Medical School, Boston, MA 02215, USA.
*   To whom correspondence should be addressed. E-mail: myaffe{at}

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Polo-like kinase is required for Golgi and bilobe biogenesis in Trypanosoma brucei.
C. L. de Graffenried, H. H. Ho, and G. Warren (2008)
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Mol. Biol. Cell 19, 2696-2707
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Science 320, 1655-1658
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N. Koida, T. Ozaki, H. Yamamoto, S. Ono, T. Koda, K. Ando, R. Okoshi, T. Kamijo, K. Omura, and A. Nakagawara (2008)
J. Biol. Chem. 283, 8555-8563
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Polo kinases regulate C. elegans embryonic polarity via binding to DYRK2-primed MEX-5 and MEX-6.
Y. Nishi, E. Rogers, S. M. Robertson, and R. Lin (2008)
Development 135, 687-697
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Z.-Q. Wu and X. Liu (2008)
PNAS 105, 1919-1924
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Phosphorylation-dependent Binding of Cyclin B1 to a Cdc6-like Domain of Human Separase.
D. Boos, C. Kuffer, R. Lenobel, R. Korner, and O. Stemmann (2008)
J. Biol. Chem. 283, 816-823
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Nucleic Acids Res. 36, D679-D683
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Cell type dependent effects of Polo-like kinase 1 inhibition compared with targeted polo box interference in cancer cell lines.
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