Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 2347K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Melixetian, M. Articles by Helin, K. Search for Related Content PubMed PubMed Citation Articles by Melixetian, M. Articles by Helin, K. Related Collections Related Article Social Bookmarking                      What's this?

¹ Department of Experimental Oncology, European Institute of Oncology, 20141 Milan, Italy
² FIRC Institute of Molecular Oncology, 20122 Milan, Italy
³ Institute of Cancer Biology, Danish Cancer Society, DK-2100 Copenhagen, Denmark
⁴ Biotech Research and Innovation Center, DK-2100 Copenhagen, Denmark

Address correspondence to Kristian Helin, Department of Experimental Oncology, European Institute of Oncology, Via Ripamonti 435, 20141 Milan, Italy. Tel.: (39) 025-748-9860. Fax: (39) 025-748-9851. email: khelin{at}ieo.it

Strict regulation of DNA replication is essential to ensure proper duplication and segregation of chromosomes during the cell cycle, as its deregulation can lead to genomic instability and cancer. Thus, eukaryotic organisms have evolved multiple mechanisms to restrict DNA replication to once per cell cycle. Here, we show that inactivation of Geminin, an inhibitor of origin licensing, leads to rereplication in human normal and tumor cells within the same cell cycle. We found a CHK1-dependent checkpoint to be activated in rereplicating cells accompanied by formation of H2AX and RAD51 nuclear foci. Abrogation of the checkpoint leads to abortive mitosis and death of rereplicated cells. In addition, we demonstrate that the induction of rereplication is dependent on the replication initiation factors CDT1 and CDC6, and independent of the functional status of p53. These data show that Geminin is required for maintaining genomic stability in human cells.

Key Words: DNA replication; genomic instability; Geminin; p53; S phase checkpoint

Abbreviations used in this paper: BAC, bacterial artificial chromosome; DSB, double-stranded DNA break; siRNA, small interfering RNA; ssDNA, single-stranded DNA.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

Related Article

Geminin halts DNA synthesis

Nicole LeBrasseur
J. Cell Biol. 2004 165: 455. [Full Text] [PDF]

This article has been cited by other articles:

• Dorn, E. S., Chastain, P. D. II, Hall, J. R., Cook, J. G. (2009). Analysis of re-replication from deregulated origin licensing by DNA fiber spreading. Nucleic Acids Res 37: 60-69 [Abstract] [Full Text]  
• Mehrotra, S., Maqbool, S. B., Kolpakas, A., Murnen, K., Calvi, B. R. (2008). Endocycling cells do not apoptose in response to DNA rereplication genotoxic stress. Genes Dev. 22: 3158-3171 [Abstract] [Full Text]  
• Ullah, Z., Kohn, M. J., Yagi, R., Vassilev, L. T., DePamphilis, M. L. (2008). Differentiation of trophoblast stem cells into giant cells is triggered by p57/Kip2 inhibition of CDK1 activity. Genes Dev. 22: 3024-3036 [Abstract] [Full Text]  
• Nishitani, H., Shiomi, Y., Iida, H., Michishita, M., Takami, T., Tsurimoto, T. (2008). CDK Inhibitor p21 Is Degraded by a Proliferating Cell Nuclear Antigen-coupled Cul4-DDB1Cdt2 Pathway during S Phase and after UV Irradiation. J. Biol. Chem. 283: 29045-29052 [Abstract] [Full Text]  
• Rakotomalala, L., Studach, L., Wang, W.-H., Gregori, G., Hullinger, R. L., Andrisani, O. (2008). Hepatitis B Virus X Protein Increases the Cdt1-to-Geminin Ratio Inducing DNA Re-replication and Polyploidy. J. Biol. Chem. 283: 28729-28740 [Abstract] [Full Text]  
• Kim, Y., Starostina, N. G., Kipreos, E. T. (2008). The CRL4Cdt2 ubiquitin ligase targets the degradation of p21Cip1 to control replication licensing. Genes Dev. 22: 2507-2519 [Abstract] [Full Text]  
• Hall, J. R., Lee, H. O., Bunker, B. D., Dorn, E. S., Rogers, G. C., Duronio, R. J., Cook, J. G. (2008). Cdt1 and Cdc6 Are Destabilized by Rereplication-induced DNA Damage. J. Biol. Chem. 283: 25356-25363 [Abstract] [Full Text]  
• Lau, E., Tsuji, T., Guo, L., Lu, S.-H., Jiang, W. (2007). The role of pre-replicative complex (pre-RC) components in oncogenesis. FASEB J. 21: 3786-3794 [Abstract] [Full Text]  
• Liu, E., Lee, A. Y.-L., Chiba, T., Olson, E., Sun, P., Wu, X. (2007). The ATR-mediated S phase checkpoint prevents rereplication in mammalian cells when licensing control is disrupted. JCB 179: 643-657 [Abstract] [Full Text]  
• Verschuren, E. W., Ban, K. H., Masek, M. A., Lehman, N. L., Jackson, P. K. (2007). Loss of Emi1-Dependent Anaphase-Promoting Complex/Cyclosome Inhibition Deregulates E2F Target Expression and Elicits DNA Damage-Induced Senescence. Mol. Cell. Biol. 27: 7955-7965 [Abstract] [Full Text]  
• Lee, A. Y.-L., Liu, E., Wu, X. (2007). The Mre11/Rad50/Nbs1 Complex Plays an Important Role in the Prevention of DNA Rereplication in Mammalian Cells. J. Biol. Chem. 282: 32243-32255 [Abstract] [Full Text]  
• Lin, J. J., Dutta, A. (2007). ATR Pathway Is the Primary Pathway for Activating G2/M Checkpoint Induction After Re-replication. J. Biol. Chem. 282: 30357-30362 [Abstract] [Full Text]  
• Hall, J. R., Kow, E., Nevis, K. R., Lu, C. K., Luce, K. S., Zhong, Q., Cook, J. G. (2007). Cdc6 Stability Is Regulated by the Huwe1 Ubiquitin Ligase after DNA Damage. Mol. Biol. Cell 18: 3340-3350 [Abstract] [Full Text]  
• Di Fiore, B., Pines, J. (2007). Emi1 is needed to couple DNA replication with mitosis but does not regulate activation of the mitotic APC/C. JCB 177: 425-437 [Abstract] [Full Text]  
• Roukos, V., Iliou, M. S., Nishitani, H., Gentzel, M., Wilm, M., Taraviras, S., Lygerou, Z. (2007). Geminin Cleavage during Apoptosis by Caspase-3 Alters Its Binding Ability to the SWI/SNF Subunit Brahma. J. Biol. Chem. 282: 9346-9357 [Abstract] [Full Text]  
• Arias, E. E., Walter, J. C. (2007). Strength in numbers: preventing rereplication via multiple mechanisms in eukaryotic cells. Genes Dev. 21: 497-518 [Abstract] [Full Text]  
• Machida, Y. J., Dutta, A. (2007). The APC/C inhibitor, Emi1, is essential for prevention of rereplication. Genes Dev. 21: 184-194 [Abstract] [Full Text]  
• Sansam, C. L., Shepard, J. L., Lai, K., Ianari, A., Danielian, P. S., Amsterdam, A., Hopkins, N., Lees, J. A. (2006). DTL/CDT2 is essential for both CDT1 regulation and the early G2/M checkpoint. Genes Dev. 20: 3117-3129 [Abstract] [Full Text]  
• Lovejoy, C. A., Lock, K., Yenamandra, A., Cortez, D. (2006). DDB1 Maintains Genome Integrity through Regulation of Cdt1. Mol. Cell. Biol. 26: 7977-7990 [Abstract] [Full Text]  
• Hara, K., Nakayama, K. I., Nakayama, K. (2006). Geminin is essential for the development of preimplantation mouse embryos. GENES CELLS 11: 1281-1293 [Abstract] [Full Text]  
• Tatsumi, Y., Sugimoto, N., Yugawa, T., Narisawa-Saito, M., Kiyono, T., Fujita, M. (2006). Deregulation of Cdt1 induces chromosomal damage without rereplication and leads to chromosomal instability. J. Cell Sci. 119: 3128-3140 [Abstract] [Full Text]  
• Gonzalez, M. A., Tachibana, K.-e K., Adams, D. J., van der Weyden, L., Hemberger, M., Coleman, N., Bradley, A., Laskey, R. A. (2006). Geminin is essential to prevent endoreduplication and to form pluripotent cells during mammalian development. Genes Dev. 20: 1880-1884 [Abstract] [Full Text]  
• Zhu, W., Dutta, A. (2006). An ATR- and BRCA1-Mediated Fanconi Anemia Pathway Is Required for Activating the G2/M Checkpoint and DNA Damage Repair upon Rereplication. Mol. Cell. Biol. 26: 4601-4611 [Abstract] [Full Text]  
• Green, B. M., Morreale, R. J., Ozaydin, B., DeRisi, J. L., Li, J. J. (2006). Genome-wide Mapping of DNA Synthesis in Saccharomyces cerevisiae Reveals That Mechanisms Preventing Reinitiation of DNA Replication Are Not Redundant. Mol. Biol. Cell 17: 2401-2414 [Abstract] [Full Text]  
• Tanny, R. E., MacAlpine, D. M., Blitzblau, H. G., Bell, S. P. (2006). Genome-wide Analysis of Re-replication Reveals Inhibitory Controls That Target Multiple Stages of Replication Initiation. Mol. Biol. Cell 17: 2415-2423 [Abstract] [Full Text]  
• Senga, T., Sivaprasad, U., Zhu, W., Park, J. H., Arias, E. E., Walter, J. C., Dutta, A. (2006). PCNA Is a Cofactor for Cdt1 Degradation by CUL4/DDB1-mediated N-terminal Ubiquitination. J. Biol. Chem. 281: 6246-6252 [Abstract] [Full Text]  
• Iizuka, M., Matsui, T., Takisawa, H., Smith, M. M. (2006). Regulation of Replication Licensing by Acetyltransferase Hbo1. Mol. Cell. Biol. 26: 1098-1108 [Abstract] [Full Text]  
• Ryu, S., Holzschuh, J., Erhardt, S., Ettl, A.-K., Driever, W. (2005). Depletion of minichromosome maintenance protein 5 in the zebrafish retina causes cell-cycle defect and apoptosis. Proc. Natl. Acad. Sci. USA 102: 18467-18472 [Abstract] [Full Text]  
• Remus, D., Blanchette, M., Rio, D. C., Botchan, M. R. (2005). CDK Phosphorylation Inhibits the DNA-binding and ATP-hydrolysis Activities of the Drosophila Origin Recognition Complex. J. Biol. Chem. 280: 39740-39751 [Abstract] [Full Text]  
• Liku, M. E., Nguyen, V. Q., Rosales, A. W., Irie, K., Li, J. J. (2005). CDK Phosphorylation of a Novel NLS-NES Module Distributed between Two Subunits of the Mcm2-7 Complex Prevents Chromosomal Rereplication. Mol. Biol. Cell 16: 5026-5039 [Abstract] [Full Text]  
• May, N. R., Thomer, M., Murnen, K. F., Calvi, B. R. (2005). Levels of the origin-binding protein Double parked and its inhibitor Geminin increase in response to replication stress. J. Cell Sci. 118: 4207-4217 [Abstract] [Full Text]  
• Archambault, V., Ikui, A. E., Drapkin, B. J., Cross, F. R. (2005). Disruption of Mechanisms That Prevent Rereplication Triggers a DNA Damage Response. Mol. Cell. Biol. 25: 6707-6721 [Abstract] [Full Text]  
• Seo, S., Herr, A., Lim, J.-W., Richardson, G. A., Richardson, H., Kroll, K. L. (2005). Geminin regulates neuronal differentiation by antagonizing Brg1 activity. Genes Dev. 19: 1723-1734 [Abstract] [Full Text]  
• Takeda, D. Y., Parvin, J. D., Dutta, A. (2005). Degradation of Cdt1 during S Phase Is Skp2-independent and Is Required for Efficient Progression of Mammalian Cells through S Phase. J. Biol. Chem. 280: 23416-23423 [Abstract] [Full Text]  
• Yanagi, K.-i., Mizuno, T., Tsuyama, T., Tada, S., Iida, Y., Sugimoto, A., Eki, T., Enomoto, T., Hanaoka, F. (2005). Caenorhabditis elegans Geminin Homologue Participates in Cell Cycle Regulation and Germ Line Development. J. Biol. Chem. 280: 19689-19694 [Abstract] [Full Text]  
• Dutta, A. (2005). Anticancer Therapy against Initiation of DNA Replication. aacredbook 2005: 243-245 [Full Text]  
• Machida, Y. J., Dutta, A. (2005). Cellular Checkpoint Mechanisms Monitoring Proper Initiation of DNA Replication. J. Biol. Chem. 280: 6253-6256 [Full Text]  
• Tsuyama, T., Tada, S., Watanabe, S., Seki, M., Enomoto, T. (2005). Licensing for DNA replication requires a strict sequential assembly of Cdc6 and Cdt1 onto chromatin in Xenopus egg extracts. Nucleic Acids Res 33: 765-775 [Abstract] [Full Text]  
• Ferenbach, A., Li, A., Brito-Martins, M., Blow, J. J. (2005). Functional domains of the Xenopus replication licensing factor Cdt1. Nucleic Acids Res 33: 316-324 [Abstract] [Full Text]  
• Yoshida, K., Takisawa, H., Kubota, Y. (2005). Intrinsic nuclear import activity of geminin is essential to prevent re-initiation of DNA replication in Xenopus eggs. GENES CELLS 10: 63-73 [Abstract] [Full Text]  
• Arias, E. E., Walter, J. C. (2005). Replication-dependent destruction of Cdt1 limits DNA replication to a single round per cell cycle in Xenopus egg extracts. Genes Dev. 19: 114-126 [Abstract] [Full Text]  
• Green, B. M., Li, J. J. (2005). Loss of Rereplication Control in Saccharomyces cerevisiae Results in Extensive DNA Damage. Mol. Biol. Cell 16: 421-432 [Abstract] [Full Text]  
• Benjamin, J. M., Torke, S. J., Demeler, B., McGarry, T. J. (2004). Geminin Has Dimerization, Cdt1-binding, and Destruction Domains That Are Required for Biological Activity. J. Biol. Chem. 279: 45957-45968 [Abstract] [Full Text]  
• Kulartz, M., Knippers, R. (2004). The Replicative Regulator Protein Geminin on Chromatin in the HeLa Cell Cycle. J. Biol. Chem. 279: 41686-41694 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents