Journal article
A Genetic Toggle for Chemical Control of Individual Plk1 Substrates
Cell chemical biology, Vol.27(3), pp.350-362.e8
03/19/2020
DOI: 10.1016/j.chembiol.2020.01.007
PMCID: PMC7239509
PMID: 32017920
Abstract
Polo-like kinase 1 has hundreds of substrates and multiple functions that operate within the ∼60 min of mitosis. Herein, we describe a chemical-genetic system that allows particular substrates to be “toggled” into or out of chemical control using engineered phosphoacceptor selectivity. Biochemical assays and phosphoproteomic analysis of mitotic cell extracts showed that Plk1S (L197F) and Plk1T (L197S/L211A) selectively phosphorylate Ser and Thr, respectively. Plk1S but not Plk1T sustains mitotic progression to anaphase, affording the opportunity to toggle substrate residues between Ser and Thr to place them under chemical control. Using this system, we evaluated Kif2b, a known substrate of Plk1 that regulates chromosome alignment. Toggling Ser to Thr on Kif2b places these phosphorylation sites under reversible chemical control, as indicated by a sharp increase in the frequency of misaligned chromosomes and prometaphase arrest. Thus, we demonstrate the ability to chemically control a single substrate by a genetic Ser/Thr toggle.
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•A new method is provided for substrate-level chemical control of kinase signaling•Chemical control is rendered by a toggle between serine and threonine residues•The chemical switch rapidly and reversibly inhibits individual phosphorylations•Control of the Plk1 substrate Kif2b is demonstrated in early mitosis
Temporal and reversible control of individual phosphorylation events is essential for a complete understanding of kinase signaling in cellular processes. Here, a chemical-genetic approach, applicable to serine-threonine kinases, achieves such control over single substrates and uncovers novel properties of Plk1 signaling through Kif2b in mitosis.
Details
- Title: Subtitle
- A Genetic Toggle for Chemical Control of Individual Plk1 Substrates
- Creators
- James M. Johnson - Highland Community College - IllinoisAlexander S. Hebert - University of Wisconsin–MadisonQuentin H. Drane - Highland Community College - IllinoisRobert F. Lera - Highland Community College - IllinoisJun Wan - University of Wisconsin–MadisonBeth A. Weaver - University of Wisconsin Carbone Cancer CenterJoshua J. Coon - University of Wisconsin–MadisonMark E. Burkard - Highland Community College - Illinois
- Resource Type
- Journal article
- Publication Details
- Cell chemical biology, Vol.27(3), pp.350-362.e8
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/j.chembiol.2020.01.007
- PMID
- 32017920
- PMCID
- PMC7239509
- ISSN
- 2451-9456
- eISSN
- 2451-9448
- Language
- English
- Date published
- 03/19/2020
- Academic Unit
- Internal Medicine
- Record Identifier
- 9984701257502771
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