Cell cycle regulation of the p34cdc2/p33cdk2-activating kinase p40MO15.
A key component of Cdc2/Cdk2-activating kinase (CAK) is p40MO15, a protein kinase subunit that phosphorylates the T161/T160 residues of p34cdc2/p33cdk2. The level and activity of p40MO15 were essentially constant during cleavage of fertilised Xenopus eggs and in growing mouse 3T3 cells, but serum starvation of these cells reduced both the level and activity of p40MO15. Although the level and activity of endogenous p40MO15 did not vary in the cell cycle, we found that bacterially expressed p40MO15 was activated more rapidly by M-phase cell extracts than by interphase cell extracts. Bacterially expressed p40MO15 was phosphorylated mainly on serine 170 (a p34cdc2 phosphorylation site) by mitotic cell extracts, but mutation of S170 to alanine did not affect the activation of p40MO15, whereas mutation of T176 (the equivalent site to T161/T160 in p34cdc2/p33cdk2) abolished the activation of P40MO15. These studies suggest that the level and activity of p40MO15 is probably not a major determinant of p34cdc2/p33cdk2 activity in the cell cycle, and that the activation of p40MO15 may require phosphorylation on T176.
Poon
R Y
RY
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, UK.
Yamashita
K
K
Howell
M
M
Ershler
M A
MA
Belyavsky
A
A
Hunt
T
T
D016475:3T3 Cells; D000409:Alanine / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology*; D002457:Cell Extracts; D018844:Cyclin-Dependent Kinases; D005982:Glutathione Transferase / Q000737:chemistry*; D007399:Interphase / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology*; D012694:Serine / Q000235:genetics; D014981:Xenopus; D000095043:Cyclin-Dependent Kinase-Activating Kinase
D002457:Cell Extracts; D011993:Recombinant Fusion Proteins; D012694:Serine; D005982:Glutathione Transferase; D017346:Protein Serine-Threonine Kinases; D018844:Cyclin-Dependent Kinases; D000409:Alanine; D000095043:Cyclin-Dependent Kinase-Activating Kinase