Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 6
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  • Aitken A . 1996 Trends Cell Biol. 6: 341–347

  • Atherton-Fessler S, Liu F, Gabrielli B, Lee M S, Peng C-Y, Piwnica-Worms H . 1994 Mol. Biol. Cell 5: 989–1001

  • Bogerd HP, Fridell RA, Benson RE, Hua J, Cullen BR . 1996 Mol. Cell. Biol. 16: 4207–4214

  • Booher RN, Holman PS, Fattaey A . 1997 J. Biol. Chem. 272: 22300–22306

  • Bunch RT, Eastman A . 1996 Clin. Cancer Res. 2: 791–797

  • Busby EC, Leistritz DF, Abraham RT, Karnitz LM, Sarkaria JN . 2000 Cancer Res. 60: 2108–2112

  • Dalal SN, Schweitzer CM, Gan J, DeCaprio J . 1999 Mol. Cell. Biol. 19: 4465–4479

  • Davezac N, Baldin V, Gabrielli B, Forrest A, Theis-Febvre N, Yashida M, Ducommun B . 2000 Oncogene 19: 2179–2185

  • Draetta G, Beach D . 1988 Cell 54: 17–26

  • Draetta G, Piwnica-Worms H, Morrison D, Druker B, Roberts T, Beach D . 1988 Nature 336: 738–744

  • Ducommun B, Draetta G, Young P, Beach D . 1990 Biochem. Biophys. Res. Commun. 167: 301–309

  • Dunphy WG, Kumagai A . 1991 Cell 67: 189–196

  • Fisher U, Huber J, Boelens WC, Mattaj IW, Luhrmann R . 1995 Cell 82: 475–483

  • Fornerod M, Ohno M, Yoshida M, Mattaj IW . 1997 Cell 90: 1051–1060

  • Fukuda M, Asano S, Nakamura T, Adachi M, Yoshida M, Yanagida M, Nishida E . 1997 Nature 390: 308–311

  • Gabrielli BG, Clark JM, McCormack AK, Ellem KA . 1997 J. Biol. Chem. 272: 28607–28614

  • Gautier J, Solomon MJ, Booher RN, Bazan JF, Kirschner MW . 1991 Cell 67: 197–211

  • Gould KL, Nurse P . 1989 Nature 342: 39–45

  • Graves PR, Yu L, Schwarz JK, Gales J, Sausville EA, O'Connor PM, Piwnica-Worms H . 2000 J. Biol. Chem. 275: 5600–5605

  • Hagting A, Karlsson C, Clute P, Jackman M, Pines J . 1998 EMBO J. 17: 4127–4138

  • He T-C, Zhou S, DaCosta LT, Yu J, Kinzler KW, Vogelstein B . 1998 Proc. Natl. Acad. Sci. 95: 2509–2514

  • Heald R, McLoughlin M, McKeon F . 1993 Cell 74: 463–474

  • Hoffmann I, Clarke PR, Marcote MJ, Karsenti E, Draetta G . 1993 EMBO J. 12: 53–63

  • Honda R, Ohba Y, Yasuda H . 1992 Biochem. Biophys. Res. Commun. 186: 1333–1338

  • Igarashi M, Nagata A, Jinno S, Suto K, Okayama H . 1991 Nature 353: 80–83

  • Izumi T, Maller JL . 1993 Mol. Biol. Cell 4: 1337–1350

  • Izumi T, Walker DH, Maller JL . 1992 Mol. Biol. Cell 3: 927–939

  • Krek W, Nigg EA . 1991 EMBO J. 10: 305–316

  • Kuang J, Ashorn CL, Gonzalez-Kuyvenhoven M, Penkala JE . 1994 Mol. Biol. Cell 5: 135–145

  • Kudo N, Khochbin S, Nishi K, Kitano K, Yanagida M, Yoshida M, Horinouchi S . 1997 J. Biol. Chem. 272: 29742–29751

  • Kumagai A, Dunphy WG . 1992 Cell 70: 139–151

  • Kumagai A, Dunphy WG . 1996 Science 273: 1377–1380

  • Kumagai A, Dunphy WG . 1999 Genes Dev. 13: 1067–1072

  • Kumagai A, Yakowec PS, Dunphy WG . 1998 Mol. Biol. Cell 9: 345–354

  • Lee MS, Ogg S, Xu M, Parker LL, Donoghue DJ, Maller JL, Piwnica-Worms H . 1992 Mol. Biol. Cell 3: 73–84

  • Li J, Meyer AN, Donoghue DJ . 1995 Mol. Biol. Cell 6: 1111–1124

  • Li J, Meyer AN, Donoghue DJ . 1997 Proc. Natl. Acad. Sci. USA 94: 502–507

  • Liu F, Stanton JJ, Wu Z, Piwnica-Worms H . 1997 Mol. Cell. Biol. 17: 571–583

  • Lopez-Girona A, Furnari B, Mondesert O, Russell P . 1999 Nature 397: 172–175

  • McGowan CH, Russell P . 1993 EMBO J. 12: 75–85

  • McGowan CH, Russell P . 1995 EMBO J. 14: 2166–2175

  • Morgan DO . 1997 Annu. Rev. Cell Dev. Biol. 13: 261–291

  • Morla AO, Draetta G, Beach D, Wang JYJ . 1989 Cell 58: 193–203

  • Mueller PR, Coleman TR, Dunphy WG . 1995a Mol. Biol. Cell 6: 119–134

  • Mueller PR, Coleman TR, Kumagai A, Dunphy WG . 1995b Science 270: 86–90

  • Muslin AJ, Tanner JW, Allen PM, Shaw AS . 1996 Cell 84: 889–897

  • Nishi K, Yoshida M, Fujiwara D, Nishikawa M, Horinouchi S, Beppu T . 1994 J. Biol. Chem. 9: 6320–6324

  • Norbury C, Blow J, Nurse P . 1991 EMBO J. 10: 3321–3329

  • Ogg S, Gabrielli B, Piwnica-Worms H . 1994 J. Biol. Chem. 269: 30461–30469

  • Ouyang B, Pan H, Lu L, Li J, Stambrook P, Li B, Dai W . 1997 J. Biol. Chem 28646–28651

  • Parker LL, Piwnica-Worms H . 1992 Science 257: 1955–1957

  • Parker LL, Sylvestre PJ, Byrnes III MJ, Liu F, Piwnica-Worms H . 1995 Proc. Natl. Acad. Sci. USA 92: 9638–9642

  • Peng C-Y, Graves PR, Thoma RS, Wu Z, Shaw A, Piwnica-Worms H . 1997 Science 277: 1501–1505

  • Pines J, Hunter T . 1994 EMBO J. 13: 3772–3781

  • Qian YW, Erickson E, Li C, Maller JL . 1998 Mol. Cell. Biol. 18: 4262–4271

  • Rittinger K, Budman J, Xu J, Volinia S, Cantley LC, Smerdon SJ, Gamblin SJ, Yaffe MB . 1999 Mol. Cell 4: 153–166

  • Sebastian B, Kakizuka A, Hunter T . 1993 Proc. Natl. Acad. Sci. USA 90: 3521–3524

  • Solomon MJ, Lee T, Kirschner MW . 1992 Mol. Biol. Cell 3: 13–27

  • Strausfeld U, Fernandez A, Capony J-P, Girard F, Lautredou N, Derancourt J, Labbe J-C, Lamb NJC . 1994 J. Biol. Chem. 269: 5989–6000

  • Strausfeld U, Labbe JC, Fesquet D, Cavadore JC, Picard A, Sadhu K, Russell P, Doree M . 1991 Nature 351: 242–245

  • Toyoshima F, Moriguchi T, Wada A, Fukuda M, Nishida E . 1998 EMBO J. 17: 2728–2735

  • Wang H, Zhang L, Liddington R, Fu H . 1998 J. Biol. Chem. 273: 16297–16304

  • Wang O, Fan S, Eastman A, Worland PJ, Sausville EA, O'Conner PM . 1996 J. Nat. Cancer Inst. 88: 956–965

  • Watanabe N, Broome M, Hunter T . 1995 EMBO J. 14: 1878–1891

  • Wen W, Meinkoth JL, Tsien RY, Taylor SS . 1995 Cell 82: 463–473

  • Yaffe MB, Rittinger K, Volinia S, Caron PR, Aikten A, Leffers H, Gamblin SJ, Smerdon SJ, Cantley LC . 1997 Cell 91: 961–971

  • Yang J, Bardes ESG, Moore JD, Brennan J, Powers MA, Kornbluth S . 1998 Genes Dev. 12: 2131–2143

  • Yang J, Kornbluth S . 1999 Trends Cell Biol. 9: 201–210

  • Yang J, Wonkler K, Yoshida M, Kornbluth S . 1999 EMBO J. 18: 2174–2183

  • Zeng Y, Forbes KC, Wu Z, Moreno S, Piwnica-Worms H, Enoch T . 1998 Nature 395: 507–510

  • Zeng Y, Piwnica-Worms H . 1999 Mol. Cell. Biol. 19: 7410–7419

Download references

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.

Author information

Author notes

  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

Authors
  1. Paul R Graves
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine M Lovly
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Geoffrey L Uy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Helen Piwnica-Worms
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204259

Keywords

Search

Advanced search

Quick links