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BOR - Papers in Press, published online ahead of print July 16, 2008.
Biol Reprod 2008, 10.1095/biolreprod.108.068205
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BIOLOGY OF REPRODUCTION 79, 991–998 (2008)
DOI: 10.1095/biolreprod.108.068205
© 2008 by the Society for the Study of Reproduction, Inc.

CDC25B Acts as a Potential Target of PRKACA in Fertilized Mouse Eggs1

Cheng Cui 3, Hongmei Zhao 4, Zhe Zhang 5, Zhihong Zong 5, Chen Feng 5, Yang Zhang 6, Xin Deng 5, Xiaoyan Xu 5, and Bingzhi Yu 2 5

Department of Physiology,3 China Medical University, Shenyang 110001, China Clinical Laboratory,4 Liao Ning Province People's Hospital, Shenyang 110003, China Department of Biochemical and Molecular Biology,5 Research Center of Medical Genome,6 Ministry of Health (MOH) of Cell Biology, China Medical University, Shenyang 110001, China

ABSTRACT

Protein kinase A (PRKACA) has been documented as a pivotal regulator in meiosis and mitosis arrest. Although our previous work has established that PRKACA regulates cell cycle progression of mouse fertilized eggs by inhibiting M-phase promoting factor (MPF), little is known about the intermediate factor between PRKACA and MPF in the mitotic cell cycle. In this study, we investigated the role of the PRKACA/CDC25B pathway on the early development of mouse fertilized eggs. Overexpression of unphosphorylatable CDC25B mutant (Cdc25b-S321A or Cdc25b-S229A/S321A) rapidly caused G2-phase eggs to enter mitosis. Microinjection of either Cdc25b-WT or Cdc25b-S229A mRNA also promoted G2/M transition, but much less efficiently than Cdc25b-S321A and Cdc25b-S229A/S321A. Moreover, mouse fertilized eggs overrode the G2 arrest by microinjection of either Cdc25b-S321A or Cdc25b-S229A/S321A mRNA, which efficiently resulted in MPF activation by directly dephosphorylating CDC2A-Tyr15, despite culture under conditions that maintained exogenous dibutyryl cAMP. Using a highly specific antibody against phospho-Ser321 of CDC25B in Western blotting, we showed that CDC25B-Ser321 was phosphorylated at the G1 and S phases, whereas Ser321 was dephosphorylated at the G2 and M phases in vivo. Our findings identify CDC25B as a potential target of PRKACA and show that PRKACA regulates G2/M transition by phosphorylating CDC25B-Ser321 but not CDC25B-Ser229 on the first mitotic division of mouse fertilized eggs..

CDC25B, M-phase promoting factor, mouse fertilized eggs, PRKACA

FOOTNOTES

1Supported by the National Nature Science Foundation of China (30570945).

Correspondence: 2Bingzhi Yu, Department of Biochemical and Molecular Biology, China Medical University, Beier Road, Heping District, Shenyang 110001, Liaoning Province, China. FAX: 0086 24 23261253; e-mail: ybzbiochem{at}yeah.net






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Copyright © 2008 by the Society for the Study of Reproduction.