Abstract
Members of the Wee family of kinases negatively regulate the cell cycle via phosphorylation of CDK1 and are considered potential drug targets. Herein, we investigated the structure-function relationship of human Wee1, Wee2, and Myt1 (PKMYT1). Purified recombinant full-length proteins and kinase domain constructs differed substantially in phosphorylation states and catalytic competency, suggesting complex mechanisms of activation. A series of crystal structures reveal unique features that distinguish Wee1 and Wee2 from Myt1 and establish the structural basis of differential inhibition by the widely used Wee1 inhibitor MK-1775. Kinome profiling and cellular studies demonstrate that, in addition to Wee1 and Wee2, MK-1775 is an equally potent inhibitor of the polo-like kinase PLK1. Several previously unrecognized inhibitors of Wee kinases were discovered and characterized. Combined, the data provide a comprehensive view on the catalytic and structural properties of Wee kinases and a framework for the rational design of novel inhibitors thereof.
MeSH terms
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Cell Cycle Proteins / antagonists & inhibitors*
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Cell Cycle Proteins / chemistry
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Cell Cycle Proteins / metabolism
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Humans
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Membrane Proteins / antagonists & inhibitors*
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Membrane Proteins / chemistry
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Membrane Proteins / metabolism
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Molecular Docking Simulation
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Nuclear Proteins / antagonists & inhibitors*
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Nuclear Proteins / chemistry
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Nuclear Proteins / metabolism
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Protein Kinase Inhibitors / chemistry*
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Protein Kinase Inhibitors / pharmacology*
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Protein Serine-Threonine Kinases / antagonists & inhibitors*
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Protein Serine-Threonine Kinases / chemistry
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Protein Serine-Threonine Kinases / metabolism
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Protein-Tyrosine Kinases / antagonists & inhibitors*
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Protein-Tyrosine Kinases / chemistry
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Protein-Tyrosine Kinases / metabolism
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Pyrazoles / chemistry
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Pyrazoles / pharmacology
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Pyrimidines / chemistry
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Pyrimidines / pharmacology
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Pyrimidinones
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Small Molecule Libraries / chemistry*
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Small Molecule Libraries / pharmacology*
Substances
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Cell Cycle Proteins
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Membrane Proteins
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Nuclear Proteins
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Protein Kinase Inhibitors
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Pyrazoles
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Pyrimidines
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Pyrimidinones
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Small Molecule Libraries
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Wee2 protein, human
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Protein-Tyrosine Kinases
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WEE1 protein, human
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PKMYT1 protein, human
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Protein Serine-Threonine Kinases
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adavosertib