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Volume 271, Number 44, Issue of November 1, 1996 pp. 27847-27854
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Sequential Dephosphorylation of p34^cdc2 on Thr-14 and Tyr-15 at the Prophase/Metaphase Transition

(Received for publication, March 28, 1996, and in revised form, August 2, 1996)

From the Centre National de la Recherche Scientifique, Station Biologique, BP 74, 29682 Roscoff cedex, France

The G₂-M transition of the cell cycle is triggered by the p34^cdc2/cyclin B kinase. During the prophase/metaphase transition, the inactive, Thr-14/Tyr-15 phosphorylated form of p34^cdc2 (T^P-Y^P) is modified to an active, Thr-14/Tyr-15 dephosphorylated form (T-Y) by the cdc25 dual-specificity phosphatase. Using highly synchronized starfish oocytes as a cellular model, we show that dephosphorylation in vivo and in vitro occurs in two steps: Thr-14 dephosphorylation precedes Tyr-15 dephosphorylation. The transient intermediate form (T-Y^P), which can be obtained in vitro by treatment of T^P-Y^P by protein phosphatase 2A, displays low but significant kinase activity. These results raise the possibility that the intermediate form T-Y^P may be involved in the autocatalytic amplification of the p34^cdc2/cyclin B complex through phosphorylation/activation of the cdc25 phosphatase and phosphorylation/inactivation of the wee1 kinase.

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