This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zeng, Y.
Right arrow Articles by Piwnica-Worms, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zeng, Y.
Right arrow Articles by Piwnica-Worms, H.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, November 1999, p. 7410-7419, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

DNA Damage and Replication Checkpoints in Fission Yeast Require Nuclear Exclusion of the Cdc25 Phosphatase via 14-3-3 Binding

Yan Zeng1 and Helen Piwnica-Worms1,2,*

Department of Cell Biology and Physiology1 and Howard Hughes Medical Institute,2 Washington University School of Medicine, St. Louis, Missouri 63110-1093

Received 10 May 1999/Returned for modification 23 June 1999/Accepted 29 July 1999

In fission yeast as well as in higher eukaryotic organisms, entry into mitosis is delayed in cells containing damaged or unreplicated DNA. This is accomplished in part by maintaining the Cdc25 phosphatase in a phosphorylated form that binds 14-3-3 proteins. In this study, we generated a mutant of fission yeast Cdc25 that is severely impaired in its ability to bind 14-3-3 proteins. Loss of both the DNA damage and replication checkpoints was observed in fission yeast cells expressing the 14-3-3 binding mutant. These findings indicate that 14-3-3 binding to Cdc25 is required for fission yeast cells to arrest their cell cycle in response to DNA damage and replication blocks. Furthermore, the 14-3-3 binding mutant localized almost exclusively to the nucleus, unlike wild-type Cdc25, which localized to both the cytoplasm and the nucleus. Nuclear accumulation of wild-type Cdc25 was observed when fission yeast cells were treated with leptomycin B, indicating that Cdc25 is actively exported from the nucleus. Nuclear exclusion of wild-type Cdc25 was observed upon overproduction of Rad 24, one of the two fission yeast 14-3-3 proteins, indicating that one function of Rad 24 is to keep Cdc25 out of the nucleus. In support of this conclusion, Rad 24 overproduction did not alter the nuclear location of the 14-3-3 binding mutant. These results indicate that 14-3-3 binding contributes to the nuclear exclusion of Cdc25 and that the nuclear exclusion of Cdc25 is required for a normal checkpoint response to both damaged and unreplicated DNA.

* Corresponding author. Mailing address: Department of Cell Biology and Physiology and Howard Hughes Medical Institute, Washington University School of Medicine, Box 8228, 660 South Euclid Ave., St. Louis, MO 63110. Phone: (314) 362-6812. Fax: (314) 362-3709. E-mail: hpiwnica{at}cellbio.wustl.edu.

Molecular and Cellular Biology, November 1999, p. 7410-7419, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Wei, S., Chen, X., Rocha, K., Epling-Burnette, P. K., Djeu, J. Y., Liu, Q., Byrd, J., Sokol, L., Lawrence, N., Pireddu, R., Dewald, G., Williams, A., Maciejewski, J., List, A. (2009). A critical role for phosphatase haplodeficiency in the selective suppression of deletion 5q MDS by lenalidomide. Proc. Natl. Acad. Sci. USA 106: 12974-12979 [Abstract] [Full Text]  
  • Jin, J., Ang, X. L., Ye, X., Livingstone, M., Harper, J. W. (2008). Differential Roles for Checkpoint Kinases in DNA Damage-dependent Degradation of the Cdc25A Protein Phosphatase. J. Biol. Chem. 283: 19322-19328 [Abstract] [Full Text]  
  • Huard, S., Elder, R. T., Liang, D., Li, G., Zhao, R. Y. (2008). Human Immunodeficiency Virus Type 1 Vpr Induces Cell Cycle G2 Arrest through Srk1/MK2-Mediated Phosphorylation of Cdc25. J. Virol. 82: 2904-2917 [Abstract] [Full Text]  
  • George, V. T., Brooks, G., Humphrey, T. C. (2007). Regulation of Cell Cycle and Stress Responses to Hydrostatic Pressure in Fission Yeast. Mol. Biol. Cell 18: 4168-4179 [Abstract] [Full Text]  
  • Dial, J. M., Petrotchenko, E. V., Borchers, C. H. (2007). Inhibition of APCCdh1 Activity by Cdh1/Acm1/Bmh1 Ternary Complex Formation. J. Biol. Chem. 282: 5237-5248 [Abstract] [Full Text]  
  • Stoica, C., Carmichael, J. B., Parker, H., Pare, J., Hobman, T. C. (2006). Interactions between the RNA Interference Effector Protein Ago1 and 14-3-3 Proteins: CONSEQUENCES FOR CELL CYCLE PROGRESSION. J. Biol. Chem. 281: 37646-37651 [Abstract] [Full Text]  
  • Pereg, Y., Lam, S., Teunisse, A., Biton, S., Meulmeester, E., Mittelman, L., Buscemi, G., Okamoto, K., Taya, Y., Shiloh, Y., Jochemsen, A. G. (2006). Differential Roles of ATM- and Chk2-Mediated Phosphorylations of Hdmx in Response to DNA Damage.. Mol. Cell. Biol. 26: 6819-6831 [Abstract] [Full Text]  
  • Gershon, E., Galiani, D., Dekel, N. (2006). Cytoplasmic polyadenylation controls cdc25B mRNA translation in rat oocytes resuming meiosis. Reproduction 132: 21-31 [Abstract] [Full Text]  
  • Kim, Y. S., Jang, S.-W., Sung, H. J., Lee, H. J., Kim, I. S., Na, D. S., Ko, J. (2005). Role of 14-3-3{eta} as a Positive Regulator of the Glucocorticoid Receptor Transcriptional Activation. Endocrinology 146: 3133-3140 [Abstract] [Full Text]  
  • Paul, A.-L., Sehnke, P. C., Ferl, R. J. (2005). Isoform-specific Subcellular Localization among 14-3-3 Proteins in Arabidopsis Seems to be Driven by Client Interactions. Mol. Biol. Cell 16: 1735-1743 [Abstract] [Full Text]  
  • Wei, J.-H., Chou, Y.-F., Ou, Y.-H., Yeh, Y.-H., Tyan, S.-W., Sun, T.-P., Shen, C.-Y., Shieh, S.-Y. (2005). TTK/hMps1 Participates in the Regulation of DNA Damage Checkpoint Response by Phosphorylating CHK2 on Threonine 68. J. Biol. Chem. 280: 7748-7757 [Abstract] [Full Text]  
  • Kino, T., Gragerov, A., Valentin, A., Tsopanomihalou, M., Ilyina-Gragerova, G., Erwin-Cohen, R., Chrousos, G. P., Pavlakis, G. N. (2005). Vpr Protein of Human Immunodeficiency Virus Type 1 Binds to 14-3-3 Proteins and Facilitates Complex Formation with Cdc25C: Implications for Cell Cycle Arrest. J. Virol. 79: 2780-2787 [Abstract] [Full Text]  
  • Dunaway, S., Liu, H.-Y., Walworth, N. C. (2005). Interaction of 14-3-3 protein with Chk1 affects localization and checkpoint function. J. Cell Sci. 118: 39-50 [Abstract] [Full Text]  
  • Jin, J., Shirogane, T., Xu, L., Nalepa, G., Qin, J., Elledge, S. J., Harper, J. W. (2003). SCF{beta}-TRCP links Chk1 signaling to degradation of the Cdc25A protein phosphatase. Genes Dev. 17: 3062-3074 [Abstract] [Full Text]  
  • Chen, M.-S., Ryan, C. E., Piwnica-Worms, H. (2003). Chk1 Kinase Negatively Regulates Mitotic Function of Cdc25A Phosphatase through 14-3-3 Binding. Mol. Cell. Biol. 23: 7488-7497 [Abstract] [Full Text]  
  • Giles, N., Forrest, A., Gabrielli, B. (2003). 14-3-3 Acts as an Intramolecular Bridge to Regulate cdc25B Localization and Activity. J. Biol. Chem. 278: 28580-28587 [Abstract] [Full Text]  
  • Perez-Hidalgo, L., Moreno, S., San-Segundo, P. A. (2003). Regulation of meiotic progression by the meiosis-specific checkpoint kinase Mek1 in fission yeast. J. Cell Sci. 116: 259-271 [Abstract] [Full Text]  
  • Zhao, H., Watkins, J. L., Piwnica-Worms, H. (2002). Disruption of the checkpoint kinase 1/cell division cycle 25A pathway abrogates ionizing radiation-induced S and G2 checkpoints. Proc. Natl. Acad. Sci. USA 99: 14795-14800 [Abstract] [Full Text]  
  • Ozoe, F., Kurokawa, R., Kobayashi, Y., Jeong, H. T., Tanaka, K., Sen, K., Nakagawa, T., Matsuda, H., Kawamukai, M. (2002). The 14-3-3 Proteins Rad24 and Rad25 Negatively Regulate Byr2 by Affecting Its Localization in Schizosaccharomyces pombe. Mol. Cell. Biol. 22: 7105-7119 [Abstract] [Full Text]  
  • Chua, G., Lingner, C., Frazer, C., Young, P. G. (2002). The sal3+ Gene Encodes an Importin-{beta} Implicated in the Nuclear Import of Cdc25 in Schizosaccharomyces pombe. Genetics 162: 689-703 [Abstract] [Full Text]  
  • Johnson, B. A., Stehn, J. R., Yaffe, M. B., Blackwell, T. K. (2002). Cytoplasmic Localization of Tristetraprolin Involves 14-3-3-dependent and -independent Mechanisms. J. Biol. Chem. 277: 18029-18036 [Abstract] [Full Text]  
  • O'Neill, T., Giarratani, L., Chen, P., Iyer, L., Lee, C.-H., Bobiak, M., Kanai, F., Zhou, B.-B., Chung, J. H., Rathbun, G. A. (2002). Determination of Substrate Motifs for Human Chk1 and hCds1/Chk2 by the Oriented Peptide Library Approach. J. Biol. Chem. 277: 16102-16115 [Abstract] [Full Text]  
  • Cueille, N., Salimova, E., Esteban, V., Blanco, M., Moreno, S., Bueno, A., Simanis, V. (2002). Flp1, a fission yeast orthologue of the S. cerevisiae CDC14 gene, is not required for cyclin degradation or rum1p stabilisation at the end of mitosis. J. Cell Sci. 114: 2649-2664 [Abstract] [Full Text]  
  • Brunet, A., Kanai, F., Stehn, J., Xu, J., Sarbassova, D., Frangioni, J. V., Dalal, S. N., DeCaprio, J. A., Greenberg, M. E., Yaffe, M. B. (2002). 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. JCB 156: 817-828 [Abstract] [Full Text]  
  • Konishi, H., Nakagawa, T., Harano, T., Mizuno, K., Saito, H., Masuda, A., Matsuda, H., Osada, H., Takahashi, T. (2002). Identification of Frequent G2 Checkpoint Impairment and a Homozygous Deletion of 14-3-3{epsilon} at 17p13.3 in Small Cell Lung Cancers. Cancer Res. 62: 271-276 [Abstract] [Full Text]  
  • Ando, T., Kawabe, T., Ohara, H., Ducommun, B., Itoh, M., Okamoto, T. (2001). Involvement of the Interaction between p21 and Proliferating Cell Nuclear Antigen for the Maintenance of G2/M Arrest after DNA Damage. J. Biol. Chem. 276: 42971-42977 [Abstract] [Full Text]  
  • Zhao, H., Piwnica-Worms, H. (2001). ATR-Mediated Checkpoint Pathways Regulate Phosphorylation and Activation of Human Chk1. Mol. Cell. Biol. 21: 4129-4139 [Abstract] [Full Text]  
  • Chen, M.-S., Hurov, J., White, L. S., Woodford-Thomas, T., Piwnica-Worms, H. (2001). Absence of Apparent Phenotype in Mice Lacking Cdc25C Protein Phosphatase. Mol. Cell. Biol. 21: 3853-3861 [Abstract] [Full Text]  
  • Zilliacus, J., Holter, E., Wakui, H., Tazawa, H., Treuter, E., Gustafsson, J.-A. (2001). Regulation of Glucocorticoid Receptor Activity by 14-3-3-Dependent Intracellular Relocalization of the Corepressor RIP140. Mol. Endocrinol. 15: 501-511 [Abstract] [Full Text]  
  • Rhind, N., Russell, P. (2001). Roles of the Mitotic Inhibitors Wee1 and Mik1 in the G2 DNA Damage and Replication Checkpoints. Mol. Cell. Biol. 21: 1499-1508 [Abstract] [Full Text]  
  • Lee, J., Kumagai, A., Dunphy, W. G. (2001). Positive Regulation of Wee1 by Chk1 and 14-3-3 Proteins. Mol. Biol. Cell 12: 551-563 [Abstract] [Full Text]  
  • Tan, S., Wang, T. S.-F. (2000). Analysis of Fission Yeast Primase Defines the Checkpoint Responses to Aberrant S Phase Initiation. Mol. Cell. Biol. 20: 7853-7866 [Abstract] [Full Text]  
  • Guo, Z., Kumagai, A., Wang, S. X., Dunphy, W. G. (2000). Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts. Genes Dev. 14: 2745-2756 [Abstract] [Full Text]  
  • Grozinger, C. M., Schreiber, S. L. (2000). Regulation of histone deacetylase 4 and 5 and transcriptional activity by 14-3- 3-dependent cellular localization. Proc. Natl. Acad. Sci. USA 10.1073/pnas.140199597v1 [Abstract] [Full Text]  
  • Pawson, T., Nash, P. (2000). Protein-protein interactions define specificity in signal transduction. Genes Dev. 14: 1027-1047 [Full Text]  
  • Graves, P. R., Yu, L., Schwarz, J. K., Gales, J., Sausville, E. A., O'Connor, P. M., Piwnica-Worms, H. (2000). The Chk1 Protein Kinase and the Cdc25C Regulatory Pathways Are Targets of the Anticancer Agent UCN-01. J. Biol. Chem. 275: 5600-5605 [Abstract] [Full Text]  
  • Rhind, N, Russell, P (2000). Chk1 and Cds1: linchpins of the DNA damage and replication checkpoint pathways. J. Cell Sci. 113: 3889-3896 [Abstract]  
  • Raleigh, J., O'Connell, M. (2000). The G(2) DNA damage checkpoint targets both Wee1 and Cdc25. J. Cell Sci. 113: 1727-1736 [Abstract]  
  • RHIND, N., BABER-FURNARI, B.A., LOPEZ-GIRONA, A., BODDY, M.N., BRONDELLO, J.-M., MOSER, B., SHANAHAN, P., BLASINA, A., MCGOWAN, C., RUSSELL, P. (2000). DNA Damage Checkpoint Control of Mitosis in Fission Yeast. Cold Spring Harb Symp Quant Biol 65: 353-360 [Abstract]  
  • Burtelow, M. A., Kaufmann, S. H., Karnitz, L. M. (2000). Retention of the Human Rad9 Checkpoint Complex in Extraction-resistant Nuclear Complexes after DNA Damage. J. Biol. Chem. 275: 26343-26348 [Abstract] [Full Text]  
  • Rauen, M., Burtelow, M. A., Dufault, V. M., Karnitz, L. M. (2000). The Human Checkpoint Protein hRad17 Interacts with the PCNA-like Proteins hRad1, hHus1, and hRad9. J. Biol. Chem. 275: 29767-29771 [Abstract] [Full Text]  
  • Gustafson, M. P., Thomas, C. F. Jr., Rusnak, F., Limper, A. H., Leof, E. B. (2001). Differential Regulation of Growth and Checkpoint Control Mediated by a Cdc25 Mitotic Phosphatase from Pneumocystis carinii. J. Biol. Chem. 276: 835-843 [Abstract] [Full Text]  
  • Curman, D., Cinel, B., Williams, D. E., Rundle, N., Block, W. D., Goodarzi, A. A., Hutchins, J. R., Clarke, P. R., Zhou, B.-B., Lees-Miller, S. P., Andersen, R. J., Roberge, M. (2001). Inhibition of the G2 DNA Damage Checkpoint and of Protein Kinases Chk1 and Chk2 by the Marine Sponge Alkaloid Debromohymenialdisine. J. Biol. Chem. 276: 17914-17919 [Abstract] [Full Text]  
  • Crawford, D. F., Piwnica-Worms, H. (2001). The G2 DNA Damage Checkpoint Delays Expression of Genes Encoding Mitotic Regulators. J. Biol. Chem. 276: 37166-37177 [Abstract] [Full Text]  
  • Grozinger, C. M., Schreiber, S. L. (2000). Regulation of histone deacetylase 4 and 5 and transcriptional activity by 14-3- 3-dependent cellular localization. Proc. Natl. Acad. Sci. USA 97: 7835-7840 [Abstract] [Full Text]  
  • Brunet, A., Kanai, F., Stehn, J., Xu, J., Sarbassova, D., Frangioni, J. V., Dalal, S. N., DeCaprio, J. A., Greenberg, M. E., Yaffe, M. B. (2002). 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. JCB 156: 817-828 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»