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Localization of human Cdc25C is regulated both by nuclear export and 14-3-3 protein binding

Oncogene volume 20, pages 1839–1851 (2001)Cite this article

Abstract

Entry into mitosis requires activation of the Cdc2 protein kinase by the Cdc25C protein phosphatase. The interactions between Cdc2 and Cdc25C are negatively regulated throughout interphase and in response to G2 checkpoint activation. This is accomplished in part by maintaining the Cdc25 phosphatase in a phosphorylated form that binds 14-3-3 proteins. Here we report that 14-3-3 binding regulates the intracellular trafficking of Cdc25C. Although primarily cytoplasmic, Cdc25C accumulated in the nuclei of leptomycin B (LMB)-treated cells, indicating that Cdc25C is actively exported out of the nucleus. A mutant of Cdc25C that is unable to bind 14-3-3 was partially nuclear in the absence of LMB and its nuclear accumulation was greatly enhanced by LMB-treatment. A nuclear export signal (NES) was identified within the amino terminus of Cdc25C. Although mutation of the NES did not effect 14-3-3 binding, it did cause nuclear accumulation of Cdc25C. These results demonstrate that 14-3-3 binding is dispensable for the nuclear export of Cdc25C. However, complete nuclear accumulation of Cdc25C required loss of both NES function and 14-3-3 binding and this was accomplished both pharmacologically and by mutation. These findings suggest that the nuclear export of Cdc25C is mediated by an intrinsic NES and that 14-3-3 binding negatively regulates nuclear import.

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Acknowledgements

We thank M Yoshida for leptomycin B and S Wente for the use of her microscope. We are grateful to C Ryan for assistance with generating recombinant adenoviruses and to J Schwarz for comments on the manuscript. J Cooper is thanked for his advice on quantitating GFP fluorescence. This work was supported in part by grants from the American Heart Association, Heartland Affiliate (to PR Graves), from the Howard Hughes Medical Institute (to GL Uy) and from the National Institutes of Health (to H Piwnica-Worms). H Piwnica-Worms is an Investigator of the Howard Hughes Medical Institute.

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  1. Paul R Graves

    Present address: Department of Pharmacology and Cancer Biology, Duke University Medical Centre, C155 LSRC Research Drive, Box 3813, Durham, 27710, NC, USA

Authors and Affiliations

  1. Department of Cell Biology and Physiology, Washington University, School of Medicine, Box 8228, 660 S Euclid Ave, St Louis, Missouri MO 63110, USA

    Paul R Graves, Christine M Lovly, Geoffrey L Uy & Helen Piwnica-Worms

  2. Howard Hughes Medical Institute, Washington University School of Medicine, St Louis, 63110, Missouri, MO, USA

    Helen Piwnica-Worms

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Graves, P., Lovly, C., Uy, G. et al. Localization of human Cdc25C is regulated both by nuclear export and 14-3-3 protein binding. Oncogene 20, 1839–1851 (2001). https://doi.org/10.1038/sj.onc.1204259

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