Periodic changes in phosphorylation of the Xenopus cdc25 phosphatase regulate its activity.
The cdc25 tyrosine phosphatase is known to activate cdc2 kinase in the G2/M transition by dephosphorylation of tyrosine 15. To determine how entry into M-phase in eukaryotic cells is controlled, we have investigated the regulation of the cdc25 protein in Xenopus eggs and oocytes. Two closely related Xenopus cdc25 genes have been cloned and sequenced and specific antibodies generated. The cdc25 phosphatase activity oscillates in both meiotic and mitotic cell cycles, being low in interphase and high in M-phase. Increased activity of cdc25 at M-phase is accompanied by increased phosphorylation that retards electrophoretic mobility in gels from 76 to 92 kDa. Treatment of cdc25 with either phosphatase 1 or phosphatase 2A removes phosphate from cdc25, reverses the mobility shift, and decreases its ability to activate cdc2 kinase. Furthermore, the addition of okadaic acid to egg extracts arrested in S-phase by aphidicolin causes phosphorylation and activation of the cdc25 protein before cyclin B/cdc2 kinase activation. These results demonstrate that the activity of the cdc25 phosphatase at the G2/M transition is directly regulated through changes in its phosphorylation state.
Izumi
T
T
Howard Hughes Medical Institute, University of Colorado, School of Medicine, Denver 80262.
Walker
D H
DH
Maller
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics; D003001:Cloning, Molecular; D016213:Cyclins / Q000378:metabolism; D004271:DNA, Fungal; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006657:Histones; D006801:Humans; D008969:Molecular Sequence Data; D009865:Oocytes; D010766:Phosphorylation; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014981:Xenopus; D020687:cdc25 Phosphatases
D016213:Cyclins; D004271:DNA, Fungal; D006657:Histones; D011506:Proteins; D016203:CDC2 Protein Kinase; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; D017027:Protein Tyrosine Phosphatases; D020687:cdc25 Phosphatases