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Mutation of a CK2 phosphorylation site in cdc25C impairs importin α/β binding and results in cytoplasmic retention

Oncogene volume 23, pages 4155–4165 (2004)Cite this article

Abstract

cdc25C is a phosphatase, which activates the mitosis-promoting factor cyclin B1/cdc2 by dephosphorylation, and thus triggers G2/M transition. The activity of cdc25C itself is controlled by phosphorylation of certain amino-acid residues, which among other things determines the subcellular localization of the enzyme. Here, we describe a new phosphorylation site at threonine 236 of cdc25C, which is phosphorylated by protein kinase CK2. This phosphorylation site is located near the nuclear localization signal (amino acids 239–245). We demonstrate that cdc25C interacts with importin β and the importin α/β heterodimer but not with importin α. We further found that a cdc25C phosphorylation mutant where threonine 236 was replaced by aspartic acid as well as cdc25C phosphorylated by CK2 binds importin β or the importin α/β heterodimer less efficiently than wild type or the corresponding alanine mutant. Furthermore, the cdc25CT236D shows a retarded uptake into the nucleus in a cell import assay. Inhibition of protein kinase CK2 enzyme activity in vivo resulted in an enhanced nuclear localization of cdc25C. Thus, phosphorylation of cdc25C at threonine 236 is an important signal for the retention of cdc25C in the cytoplasm.

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Acknowledgements

We thank Jürgen Günther for technical assistance, Marianne Buchholz for help with the manuscript and Dr Andrea Krempler for careful reading of the manuscript and many helpful suggestions. We thank Dr David Jans for kindly providing importin α and importin β constructs, Dr Hans Stahl for purified SV40 large T antigen and Dr Wolfgang Nastainczyk for the synthesis of the peptide filters. This work was supported by a grant from Deutsche Forschungsgemeinschaft Mo 309/11-3.

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  1. Universität des Saarlandes, Medizinische Biochemie und Molekularbiologie, Gebäude 44, D-66421, Homburg, Germany

    Sandra L Schwindling, Andreas Noll, Mathias Montenarh & Claudia Götz

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  1. Sandra L Schwindling
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  2. Andreas Noll
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  3. Mathias Montenarh
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Correspondence to Claudia Götz.

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Schwindling, S., Noll, A., Montenarh, M. et al. Mutation of a CK2 phosphorylation site in cdc25C impairs importin α/β binding and results in cytoplasmic retention. Oncogene 23, 4155–4165 (2004). https://doi.org/10.1038/sj.onc.1207566

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