A Genetic Toggle for Chemical Control of Individual Plk1 Substrates. Issue 3 (19th March 2020)

• Record Type: Journal Article
• Authors: Johnson, James M.; Hebert, Alexander S.; Drane, Quentin H.; Lera, Robert F.; Wan, Jun; Weaver, Beth A.; Coon, Joshua J.; Burkard, Mark E.
• Is Part Of: Cell chemical biology. Volume 27:Issue 3(2020)
• Page Start: 350
• Page End: 362.e8
• Publication Date: 2020-03-19
• Subjects: chemical biology -- kinase -- genetic toggle -- phosphoacceptor selectivity -- chemical-genetics -- mitosis -- phosphorylation -- Kif2b; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2020.01.007
• Languages: English
• ISSNs: 2451-9456

Abstract

Summary: 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 Plk1 S (L197F) and Plk1 T (L197S/L211A) selectively phosphorylate Ser and Thr, respectively. Plk1 S but not Plk1 T 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. Graphical Abstract: Highlights: 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 Abstract : 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.