A Coupled Chemical-Genetic and Bioinformatic Approach to Polo-like Kinase Pathway Exploration
| Autor: | Michael S. Cohen, Kevan M. Shokat, Jennifer Snead, David H. Randle, David O. Morgan, Jack Taunton, Michael B. Yaffe, Matt Sullivan, Chao Zhang, Drew M. Lowery |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Saccharomyces cerevisiae Proteins
Clinical Biochemistry Cell Cycle Proteins macromolecular substances Mitogen-activated protein kinase kinase Protein Serine-Threonine Kinases Biochemistry Spindle pole body Article MAP2K7 Substrate Specificity 03 medical and health sciences 0302 clinical medicine Drug Discovery Integrin-linked kinase ASK1 Molecular Biology Alleles 030304 developmental biology Pharmacology 0303 health sciences biology MAP kinase kinase kinase Cyclin-dependent kinase 2 Computational Biology General Medicine Cell biology enzymes and coenzymes (carbohydrates) CHEMBIO SIGNALING biology.protein Cyclin-dependent kinase complex Molecular Medicine Protein Kinases 030217 neurology & neurosurgery |
| Zdroj: | Chemistry & Biology. (11):1261-1272 |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/j.chembiol.2007.09.011 |
| Popis: | 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. Monospecific pharmacological inhibition of budding yeast Polo-like kinase, Cdc5, resulted in a misaligned pre-anaphase spindle and subsequently delayed anaphase nuclear migration, revealing a novel 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. |
| Databáze: | OpenAIRE |
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