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[PKA-regulated phosphorylation status of S149 and S321 sites of CDC25B inhibits mitosis of fertilized mouse eggs.] [Article in Chinese]

XIAO Jian-Ying, LIU Chao, SUN Xiao-Han, YU Bing-Zhi^*

Department of Biochemical and Molecular Biology, China Medical University, Shenyang 110001, China； Department of Biochemical and Molecular Biology, Liaoning Medial University, Jinzhou 121001, China

Abstract

To further test whether protein kinase A (PKA) can affect the mitotic cell cycle, one-cell stage mouse embryos at S phase (22 h after hCG injection) were incubated in M16 medium containing various concentrations of H-89, a PKA inhibitor. With increasing concentrations of H-89 (0–50 μmol/L), the G2 phase of eggs was decreased and the cleavage rate was accelerated. A concentration of 40 μmol/L H-89 led to all of the mouse eggs entering the M phase of mitosis. Furthermore, to study the role of PKA in regulating the phosphorylation status of S149 and S321 sites of cell division cycle 25B (CDC25B) on one-cell stage fertilized mouse eggs, pBSK-CDC25B-WT, pBSK-CDC25B-S149A, pBSK-CDC25B-S321A and pBSK-CDC25B-S149A/S321A were transcribed into mRNAs in vitro, then mRNAs were microinjected into S phase of mouse fertilized eggs and cultured in M16 medium pretreated with H-89. Then, the cleavage of fertilized eggs, maturation promoting factor (MPF) activity and phosphorylation status of CDC2-Tyr15 were observed. In the presence of 40 μmol/L H-89, the cleavage rate of fertilized eggs in CDC25B-S/A-mRNAs and CDC25B-WT-mRNA injected groups was significantly higher than that in the control groups, and the peak of MPF activity appeared in the CDC25B-S/A-mRNAs and CDC25B-WT-mRNA injected groups earlier than that in the control groups. CDC2-Tyr15 phosphorylation state was consistent with MPF activity. In conclusion, the present study suggests that PKA regulates the early development of mouse embryos by phosphorylation of S149 and S321 of CDC25B, which plays an important role in the regulation of G2/M transition in the mitotic cell cycle of fertilized mouse eggs.

Key words: cell division cycle 25B; protein kinase A; maturation promoting factor; H-89; fertilized mouse eggs

Received: 2011-07-30　　Accepted: 2011-09-15

Corresponding author: 于秉治　　E-mail: ybzbiochem@yeah.net

Citing This Article：

XIAO Jian-Ying, LIU Chao, SUN Xiao-Han, YU Bing-Zhi. [PKA-regulated phosphorylation status of S149 and S321 sites of CDC25B inhibits mitosis of fertilized mouse eggs.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (1): 33-40 (in Chinese with English abstract).