14-3-3 binding regulates catalytic activity of human Wee1 kinase.
The mitotic inducer Cdc2 is negatively regulated, in part, by phosphorylation on tyrosine 15. Human Wee1 is a tyrosine-specific protein kinase that phosphorylates Cdc2 on tyrosine 15. Human Wee1 is subject to multiple levels of regulation including reversible phosphorylation, proteolysis, and protein-protein interactions. Here we have investigated the contributions made by 14-3-3 binding to human Wee1 regulation and function. We report that the interactions of 14-3-3 proteins with human Wee1 are reduced during mitosis and are stable in the presence of the protein kinase inhibitor UCN-01. A mutant of Wee1 that is incapable of binding to 14-3-3 proteins has lower enzymatic activity, and this likely accounts for its reduced potency relative to wild-type Wee1 in inducing a G(2) cell cycle delay when overproduced in vivo. These findings indicate that 14-3-3 proteins function as positive regulators of the human Wee1 protein kinase.
Rothblum-Oviatt
C J
CJ
Department of Cell Biology and Physiology, Howard Hughes Medical Institute, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110, USA.
Ryan
C E
CE
Piwnica-Worms
H
H
D048948:14-3-3 Proteins; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002384:Catalysis; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D004926:Escherichia coli / Q000378:metabolism; D016195:G2 Phase; D006367:HeLa Cells; D006801:Humans; D008938:Mitosis; D009687:Nuclear Proteins; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism* / Q000502:physiology*
D048948:14-3-3 Proteins; D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D014446:Tyrosine 3-Monooxygenase; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human