14-3-3 binding regulates catalytic activity of human Wee1 kinase

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C J Rothblum-Oviatt1, C E Ryan, H Piwnica-Worms

  • 1Department of Cell Biology and Physiology, Howard Hughes Medical Institute, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110, USA.

Cell Growth & Differentiation : the Molecular Biology Journal of the American Association for Cancer Research
|December 26, 2001
PubMed

Insights

14-3-3 proteins positively regulate human Wee1, a key kinase in cell cycle control. Reduced binding during mitosis and impaired function of non-binding Wee1 mutants highlight 14-3-3's crucial role.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The cell cycle is tightly regulated by protein kinases, including Cdc2.
  • Wee1 kinase phosphorylates Cdc2 at tyrosine 15, inhibiting its activity and inducing cell cycle arrest.
  • Human Wee1 is regulated by phosphorylation, proteolysis, and protein interactions.

Purpose of the Study:

  • To investigate the role of 14-3-3 protein interactions in regulating human Wee1 kinase activity and function.
  • To determine how 14-3-3 binding to Wee1 changes during the cell cycle.

Main Methods:

  • Studied 14-3-3 binding to human Wee1 using biochemical assays.
  • Utilized a Wee1 mutant unable to bind 14-3-3 proteins.
  • Assessed enzymatic activity and cell cycle delay (G2) in vivo.

Main Results:

  • 14-3-3 binding to human Wee1 decreases during mitosis.
  • 14-3-3 binding remains stable when treated with the protein kinase inhibitor UCN-01.
  • Wee1 mutants lacking 14-3-3 binding exhibit reduced enzymatic activity and potency in inducing G2 delay.

Conclusions:

  • 14-3-3 proteins act as positive regulators of human Wee1 kinase activity.
  • 14-3-3 binding is essential for Wee1's function in cell cycle progression.

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