Ser149 is Another Potential PKA Phosphorylation Target of Cdc25B in G2/M Transition of Fertilized Mouse Eggs

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2011 Jan 8

PMID 21212267

Citations 8

Authors

Jianying Xiao

Chao Liu

Junjie Hou

Cheng Cui

Didi Wu

Huiyu Fan

Xiaohan Sun

Jun Meng

Fuquan Yang

Enhua Wang

Bingzhi Yu

Affiliations

Soon will be listed here.

Abstract

It is well documented that protein kinase A (PKA) acts as a negative regulator of M phase promoting factor (MPF) by phosphorylating cell division cycle 25 homolog B (Cdc25B) in mammals. However, the molecular mechanism remains unclear. In this study, we identified PKA phosphorylation sites in vitro by LC-MS/MS analysis, including Ser(149), Ser(229), and Ser(321) of Cdc25B, and explored the role of Ser(149) in G(2)/M transition of fertilized mouse eggs. The results showed that the overexpressed Cdc25B-S149A mutant initiated efficient MPF activation by direct dephosphorylation of Cdc2-Tyr(15), resulting in triggering mitosis prior to Cdc25B-WT. Conversely, overexpression of the phosphomimic Cdc25B-S149D mutant showed no significant difference in comparison with the control groups. Furthermore, we found that Cdc25B-Ser(149) was phosphorylated at G(1) and S phases, whereas dephosphorylated at G(2) and M phases, and the phosphorylation of Cdc25B-Ser(149) was modulated by PKA in vivo. In addition, we examined endogenous and exogenous Cdc25B, which were expressed mostly in the cytoplasm at the G(1) and S phases and translocated to the nucleus at the G(2) phase. Collectively, our findings provide evidence that Ser(149) may be another potential PKA phosphorylation target of Cdc25B in G(2)/M transition of fertilized mouse eggs and Cdc25B as a direct downstream substrate of PKA in mammals, which plays important roles in the regulation of early development of mouse embryos.

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References

Giles N, Forrest A, Gabrielli B . 14-3-3 acts as an intramolecular bridge to regulate cdc25B localization and activity. J Biol Chem. 2003; 278(31):28580-7. DOI: 10.1074/jbc.M304027200. View

Alonso A, Sasin J, Bottini N, Friedberg I, Friedberg I, Osterman A . Protein tyrosine phosphatases in the human genome. Cell. 2004; 117(6):699-711. DOI: 10.1016/j.cell.2004.05.018. View

Boutros R, Lobjois V, Ducommun B . CDC25 phosphatases in cancer cells: key players? Good targets?. Nat Rev Cancer. 2007; 7(7):495-507. DOI: 10.1038/nrc2169. View

Nilsson I, Hoffmann I . Cell cycle regulation by the Cdc25 phosphatase family. Prog Cell Cycle Res. 2000; 4:107-14. DOI: 10.1007/978-1-4615-4253-7_10. View

Schultz R, Montgomery R, Belanoff J . Regulation of mouse oocyte meiotic maturation: implication of a decrease in oocyte cAMP and protein dephosphorylation in commitment to resume meiosis. Dev Biol. 1983; 97(2):264-73. DOI: 10.1016/0012-1606(83)90085-4. View

Chen J, Hudson E, Chi M, Chang A, Moley K, Hardie D . AMPK regulation of mouse oocyte meiotic resumption in vitro. Dev Biol. 2006; 291(2):227-38. DOI: 10.1016/j.ydbio.2005.11.039. View

Gallicano G, McGaughey R, Capco D . Activation of protein kinase C after fertilization is required for remodeling the mouse egg into the zygote. Mol Reprod Dev. 1997; 46(4):587-601. DOI: 10.1002/(SICI)1098-2795(199704)46:4<587::AID-MRD16>3.0.CO;2-T. View

Strausfeld U, Labbe J, Fesquet D, Cavadore J, Picard A, Sadhu K . Dephosphorylation and activation of a p34cdc2/cyclin B complex in vitro by human CDC25 protein. Nature. 1991; 351(6323):242-5. DOI: 10.1038/351242a0. View

Boutros R, Dozier C, Ducommun B . The when and wheres of CDC25 phosphatases. Curr Opin Cell Biol. 2006; 18(2):185-91. DOI: 10.1016/j.ceb.2006.02.003. View

10.

Studier F . Protein production by auto-induction in high density shaking cultures. Protein Expr Purif. 2005; 41(1):207-34. DOI: 10.1016/j.pep.2005.01.016. View

11.

Cui C, Zhao H, Zhang Z, Zong Z, Feng C, Zhang Y . CDC25B acts as a potential target of PRKACA in fertilized mouse eggs. Biol Reprod. 2008; 79(5):991-8. DOI: 10.1095/biolreprod.108.068205. View

12.

Jeannine Lincoln A, Wickramasinghe D, Stein P, Schultz R, Palko M, De Miguel M . Cdc25b phosphatase is required for resumption of meiosis during oocyte maturation. Nat Genet. 2002; 30(4):446-9. DOI: 10.1038/ng856. View

13.

Eyers P, Liu J, Hayashi N, Lewellyn A, Gautier J, Maller J . Regulation of the G(2)/M transition in Xenopus oocytes by the cAMP-dependent protein kinase. J Biol Chem. 2005; 280(26):24339-46. DOI: 10.1074/jbc.M412442200. View

14.

Lammer C, Wagerer S, Saffrich R, Mertens D, Ansorge W, Hoffmann I . The cdc25B phosphatase is essential for the G2/M phase transition in human cells. J Cell Sci. 1998; 111 ( Pt 16):2445-53. DOI: 10.1242/jcs.111.16.2445. View

15.

Lobjois V, Jullien D, Bouche J, Ducommun B . The polo-like kinase 1 regulates CDC25B-dependent mitosis entry. Biochim Biophys Acta. 2009; 1793(3):462-8. DOI: 10.1016/j.bbamcr.2008.12.015. View

16.

Li X, Gerber S, Rudner A, Beausoleil S, Haas W, Villen J . Large-scale phosphorylation analysis of alpha-factor-arrested Saccharomyces cerevisiae. J Proteome Res. 2007; 6(3):1190-7. DOI: 10.1021/pr060559j. View

17.

Eng J, McCormack A, Yates J . An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Soc Mass Spectrom. 2013; 5(11):976-89. DOI: 10.1016/1044-0305(94)80016-2. View

18.

Grieco D, Avvedimento E, GOTTESMAN M . A role for cAMP-dependent protein kinase in early embryonic divisions. Proc Natl Acad Sci U S A. 1994; 91(21):9896-900. PMC: 44924. DOI: 10.1073/pnas.91.21.9896. View

19.

Rudolph J . Cdc25 phosphatases: structure, specificity, and mechanism. Biochemistry. 2007; 46(12):3595-604. DOI: 10.1021/bi700026j. View

20.

Solc P, Saskova A, Baran V, Kubelka M, Schultz R, Motlik J . CDC25A phosphatase controls meiosis I progression in mouse oocytes. Dev Biol. 2008; 317(1):260-9. PMC: 2430978. DOI: 10.1016/j.ydbio.2008.02.028. View