A Coupled Chemical-Genetic and Bioinformatic Approach to Polo-like Kinase Pathway Exploration

Jennifer L. Snead, Matthew Sullivan, Drew M. Lowery, Michael S. Cohen, Chao Zhang, David H. Randle, Jack Taunton, Michael B. Yaffe, David O. Morgan, Kevan M. Shokat

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

Protein phosphorylation is a ubiquitous mechanism for cellular signal propagation, and signaling network complexity presents a challenge to protein kinase substrate identification. Few targets of Polo-like kinases are known, despite their significant role in coordinating cell-cycle progression. Here, we combine chemical-genetic, bioinformatic, and proteomic tools for Polo-like kinase substrate identification. Specific pharmacological inhibition of budding yeast Polo-like kinase, Cdc5, resulted in a misaligned preanaphase spindle and subsequently delayed anaphase nuclear migration, revealing a Cdc5 function. A cellular screen for Cdc5 substrates identified Spc72, a spindle pole body (SPB) component and microtubule anchor required for nuclear positioning. Spc72 bound to the Cdc5 PBD in a mitosis-specific manner, was phosphorylated by Cdc5 in vitro, and demonstrated a loss of mitotic phosphorylation in vivo upon Cdc5 inhibition. Finally, an examination of Cdc5 binding by SPB-localized proteins expanded our knowledge of Cdc5 function at the SPB.

Original languageEnglish (US)
Pages (from-to)1261-1272
Number of pages12
JournalChemistry and Biology
Volume14
Issue number11
DOIs
StatePublished - Nov 26 2007

Keywords

  • CHEMBIO
  • SIGNALING

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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    Snead, J. L., Sullivan, M., Lowery, D. M., Cohen, M. S., Zhang, C., Randle, D. H., Taunton, J., Yaffe, M. B., Morgan, D. O., & Shokat, K. M. (2007). A Coupled Chemical-Genetic and Bioinformatic Approach to Polo-like Kinase Pathway Exploration. Chemistry and Biology, 14(11), 1261-1272. https://doi.org/10.1016/j.chembiol.2007.09.011