Two distinct mechanisms for negative regulation of the Wee1 protein kinase.
The Wee1 protein kinase negatively regulates the entry into mitosis by catalyzing the inhibitory tyrosine phosphorylation of the Cdc2 protein. To examine the potential mechanisms for Wee1 regulation during the cell cycle, we have introduced a recombinant form of the fission yeast Wee1 protein kinase into Xenopus egg extracts. We find that the Wee1 protein undergoes dramatic changes in its phosphorylation state and kinase activity during the cell cycle. The Wee1 protein oscillates between an underphosphorylated 107 kDa form during interphase and a hyperphosphorylated 170 kDa version at mitosis. The mitosis-specific hyperphosphorylation of the Wee1 protein results in a substantial reduction in its activity as a Cdc2-specific tyrosine kinase. This phosphorylation occurs in the N-terminal region of the protein that lies outside the C-terminal catalytic domain, which was recently shown to be a substrate for the fission yeast Nim1 protein kinase. These experiments demonstrate the existence of a Wee1 regulatory system, consisting of both a Wee1-inhibitory kinase and a Wee1-stimulatory phosphatase, which controls the phosphorylation of the N-terminal region of the Wee1 protein. Moreover, these findings indicate that there are apparently two potential mechanisms for negative regulation of the Wee1 protein, one involving phosphorylation of its C-terminal domain by the Nim1 protein and the other involving phosphorylation of its N-terminal region by a different kinase.
Tang
Z
Z
Division of Biology, California Institute of Technology, Pasadena 91125.
Coleman
T R
TR
Dunphy
W G
WG
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D018797:Cell Cycle Proteins; D002460:Cell Line; D016213:Cyclins / Q000235:genetics / Q000378:metabolism; D005260:Female; D006706:Homeostasis; D006801:Humans; D007313:Insecta; D008938:Mitosis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D010063:Ovum / Q000502:physiology; D010766:Phosphorylation; D019000:Phosphotyrosine; D011494:Protein Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases; D015183:Restriction Mapping; D012568:Schizosaccharomyces / Q000201:enzymology; D029702:Schizosaccharomyces pombe Proteins; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D014981:Xenopus; D029867:Xenopus Proteins
D018797:Cell Cycle Proteins; D016213:Cyclins; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D029702:Schizosaccharomyces pombe Proteins; D029867:Xenopus Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D011494:Protein Kinases; C530005:WEE1 protein, Xenopus; C068689:wee1 protein, S pombe; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; D016203:CDC2 Protein Kinase