Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Logo for

J. Cell Biol. 156 (2): 249-260

Copyright © 2002 by the Rockefeller University Press.

Article

The checkpoint protein Chfr is a ligase that ubiquitinates Plk1 and inhibits Cdc2 at the G2 to M transition

Dongmin Kang, James Chen, Jim Wong, and Guowei Fang

Department of Biological Sciences, Stanford University, Stanford, CA 94305

Address correspondence to Guowei Fang, Department of Biological Sciences, Stanford University, 385 Serra Mall, MC-5020, Stanford, CA 94305-5020. Tel.: (650) 725-2762. Fax: (650) 725-5807. E-mail: gwfang{at}stanford.edu

Abstract: The checkpoint protein Chfr delays entry into mitosis, in the presence of mitotic stress (Scolnick, D.M., and T.D. Halazonetis. 2000. Nature. 406:430–435). We show here that Chfr is a ubiquitin ligase, both in vitro and in vivo. When transfected into HEK293T cells, Myc–Chfr promotes the formation of high molecular weight ubiquitin conjugates. The ring finger domain in Chfr is required for the ligase activity; this domain auto-ubiquitinates, and mutations of conserved residues in this domain abolish the ligase activity. Using Xenopus cell-free extracts, we demonstrated that Chfr delays the entry into mitosis by negatively regulating the activation of the Cdc2 kinase at the G2–M transition. Specifically, the Chfr pathway prolongs the phosphorylated state of tyrosine 15 in Cdc2. The Chfr-mediated cell cycle delay requires ubiquitin-dependent protein degradation, because inactivating mutations in Chfr, interference with poly-ubiquitination, and inhibition of proteasomes all abolish this delay in mitotic entry. The direct target of the Chfr pathway is Polo-like kinase 1 (Plk1). Ubiquitination of Plk1 by Chfr delays the activation of the Cdc25C phosphatase and the inactivation of the Wee1 kinase, leading to a delay in Cdc2 activation. Thus, the Chfr pathway represents a novel checkpoint pathway that regulates the entry into mitosis by ubiquitin-dependent proteolysis.

Key Words: Chfr; Cdc2; Plk1; mitosis; ubiquitin protein ligase

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
CHFR Protein Expression Predicts Outcomes to Taxane-Based First Line Therapy in Metastatic NSCLC.
R. N. Pillai, S. A. Brodie, G. L. Sica, Y. Shaojin, G. Li, D. C. Nickleach, L. Yuan, V. A. Varma, D. Bonta, J. G. Herman, et al. (2013)
Clin. Cancer Res. 19, 1603-1611
   Abstract »    Full Text »    PDF »
Decoding Ubiquitin for Mitosis.
S. Fournane, K. Krupina, C. Kleiss, and I. Sumara (2013)
Genes & Cancer
   Abstract »    Full Text »    PDF »
Curcumin-induced mitotic arrest is characterized by spindle abnormalities, defects in chromosomal congression and DNA damage.
L. M. Blakemore, C. Boes, R. Cordell, and M. M. Manson (2013)
Carcinogenesis 34, 351-360
   Abstract »    Full Text »    PDF »
Dnt1 acts as a mitotic inhibitor of the spindle checkpoint protein dma1 in fission yeast.
Y. Wang, W.-z. Li, A. E. Johnson, Z.-q. Luo, X.-l. Sun, A. Feoktistova, W. H. McDonald, I. McLeod, J. R. Yates III, K. L. Gould, et al. (2012)
Mol. Biol. Cell 23, 3348-3356
   Abstract »    Full Text »    PDF »
CHFR Protein Regulates Mitotic Checkpoint by Targeting PARP-1 Protein for Ubiquitination and Degradation.
L. Kashima, M. Idogawa, H. Mita, M. Shitashige, T. Yamada, K. Ogi, H. Suzuki, M. Toyota, H. Ariga, Y. Sasaki, et al. (2012)
J. Biol. Chem. 287, 12975-12984
   Abstract »    Full Text »    PDF »
The Auto-ubiquitylation of E3 Ubiquitin-protein Ligase Chfr at G2 Phase Is Required for Accumulation of Polo-like Kinase 1 and Mitotic Entry in Mammalian Cells.
J.-S. Kim, Y.-Y. Park, S.-Y. Park, H. Cho, D. Kang, and H. Cho (2011)
J. Biol. Chem. 286, 30615-30623
   Abstract »    Full Text »    PDF »
The Stil protein regulates centrosome integrity and mitosis through suppression of Chfr.
A. Castiel, M. M. Danieli, A. David, S. Moshkovitz, P. D. Aplan, I. R. Kirsch, M. Brandeis, A. Kramer, and S. Izraeli (2011)
J. Cell Sci. 124, 532-539
   Abstract »    Full Text »    PDF »
Structural Basis of Poly(ADP-ribose) Recognition by the Multizinc Binding Domain of Checkpoint with Forkhead-associated and RING Domains (CHFR).
J. Oberoi, M. W. Richards, S. Crumpler, N. Brown, J. Blagg, and R. Bayliss (2010)
J. Biol. Chem. 285, 39348-39358
   Abstract »    Full Text »    PDF »
Safeguarding Entry into Mitosis: the Antephase Checkpoint.
C. F. Chin and F. M. Yeong (2010)
Mol. Cell. Biol. 30, 22-32
   Abstract »    Full Text »    PDF »
Molecular Distinctions between Aurora A and B: A Single Residue Change Transforms Aurora A into Correctly Localized and Functional Aurora B.
F. Hans, D. A. Skoufias, S. Dimitrov, and R. L. Margolis (2009)
Mol. Biol. Cell 20, 3491-3502
   Abstract »    Full Text »    PDF »
Polo-like kinase 1 is overexpressed in acute myeloid leukemia and its inhibition preferentially targets the proliferation of leukemic cells.
A. G. Renner, C. Dos Santos, C. Recher, C. Bailly, L. Creancier, A. Kruczynski, B. Payrastre, and S. Manenti (2009)
Blood 114, 659-662
   Abstract »    Full Text »    PDF »
Functional Interaction between Chfr and Kif22 Controls Genomic Stability.
S. Maddika, S. M.-H. Sy, and J. Chen (2009)
J. Biol. Chem. 284, 12998-13003
   Abstract »    Full Text »    PDF »
Persistent FoxE3 Expression Blocks Cytoskeletal Remodeling and Organelle Degradation during Lens Fiber Differentiation.
H. Landgren, A. Blixt, and P. Carlsson (2008)
Invest. Ophthalmol. Vis. Sci. 49, 4269-4277
   Abstract »    Full Text »    PDF »
G2E3 Is a Dual Function Ubiquitin Ligase Required for Early Embryonic Development.
W. S. Brooks, E. S. Helton, S. Banerjee, M. Venable, L. Johnson, T. R. Schoeb, R. A. Kesterson, and D. F. Crawford (2008)
J. Biol. Chem. 283, 22304-22315
   Abstract »    Full Text »    PDF »
The Cech Symposium: A celebration of 25 years of ribozymes, 10 years of TERT, and 60 years of Tom.
Q. Vicens, M. A. Allen, S. D. Gilbert, B. Reznik, A. R. Gooding, and R. T. Batey (2008)
RNA 14, 397-403
   Abstract »    Full Text »    PDF »
Defective in Mitotic Arrest 1/Ring Finger 8 Is a Checkpoint Protein That Antagonizes the Human Mitotic Exit Network.
R. L. Tuttle, J. Bothos, M. K. Summers, F. C. Luca, and T. D. Halazonetis (2007)
Mol. Cancer Res. 5, 1304-1311
   Abstract »    Full Text »    PDF »
Altered Expression of the Early Mitotic Checkpoint Protein, CHFR, in Breast Cancers: Implications for Tumor Suppression.
L. M. Privette, M. E. Gonzalez, L. Ding, C. G. Kleer, and E. M. Petty (2007)
Cancer Res. 67, 6064-6074
   Abstract »    Full Text »    PDF »
The tumor suppressor CYLD regulates entry into mitosis.
F. Stegmeier, M. E. Sowa, G. Nalepa, S. P. Gygi, J. W. Harper, and S. J. Elledge (2007)
PNAS 104, 8869-8874
   Abstract »    Full Text »    PDF »
A census of mitotic cancer genes: new insights into tumor cell biology and cancer therapy.
I. Perez de Castro, G. de Carcer, and M. Malumbres (2007)
Carcinogenesis 28, 899-912
   Abstract »    Full Text »    PDF »
Human T-Cell Leukemia Virus Oncoprotein Tax Represses Nuclear Receptor-Dependent Transcription by Targeting Coactivator TAX1BP1.
K.-T. Chin, A. C.S. Chun, Y.-P. Ching, K.-T. Jeang, and D.-Y. Jin (2007)
Cancer Res. 67, 1072-1081
   Abstract »    Full Text »    PDF »
Genetic and Expression Aberrations of E3 Ubiquitin Ligases in Human Breast Cancer.
C. Chen, A. K. Seth, and A. E. Aplin (2006)
Mol. Cancer Res. 4, 695-707
   Abstract »    Full Text »    PDF »
Hamartin, the tuberous sclerosis complex 1 gene product, interacts with polo-like kinase 1 in a phosphorylation-dependent manner.
A. Astrinidis, W. Senapedis, and E. P. Henske (2006)
Hum. Mol. Genet. 15, 287-297
   Abstract »    Full Text »    PDF »
Down-Regulation of DNA Topoisomerase II{alpha} Leads to Prolonged Cell Cycle Transit in G2 and Early M Phases and Increased Survival to Microtubule-Interacting Agents.
A. Skladanowski, M.-G. Come, M. Sabisz, A. E. Escargueil, and A. K. Larsen (2005)
Mol. Pharmacol. 68, 625-634
   Abstract »    Full Text »    PDF »
CHFR promoter hypermethylation in colon cancer correlates with the microsatellite instability phenotype.
J. C. Brandes, M. van Engeland, K. A.D. Wouters, M. P. Weijenberg, and J. G. Herman (2005)
Carcinogenesis 26, 1152-1156
   Abstract »    Full Text »    PDF »
Deficiency in SNM1 Abolishes an Early Mitotic Checkpoint Induced by Spindle Stress.
S. Akhter, C. T. Richie, J. M. Deng, E. Brey, X. Zhang, C. Patrick Jr., R. R. Behringer, and R. J. Legerski (2004)
Mol. Cell. Biol. 24, 10448-10455
   Abstract »    Full Text »    PDF »
Cdc123 and Checkpoint Forkhead Associated with RING Proteins Control the Cell Cycle by Controlling eIF2{gamma} Abundance.
P. Bieganowski, K. Shilinski, P. N. Tsichlis, and C. Brenner (2004)
J. Biol. Chem. 279, 44656-44666
   Abstract »    Full Text »    PDF »
Chfr acts with the p38 stress kinases to block entry to mitosis in mammalian cells.
T. Matsusaka and J. Pines (2004)
J. Cell Biol. 166, 507-516
   Abstract »    Full Text »    PDF »
Functional Characterization of Dma1 and Dma2, the Budding Yeast Homologues of Schizosaccharomyces pombe Dma1 and Human Chfr.
R. Fraschini, D. Bilotta, G. Lucchini, and S. Piatti (2004)
Mol. Biol. Cell 15, 3796-3810
   Abstract »    Full Text »    PDF »
Identification of human polo-like kinase 1 as a potential therapeutic target in pancreatic cancer.
P. J. Gray Jr, D. J. Bearss, H. Han, R. Nagle, M.-S. Tsao, N. Dean, and D. D. Von Hoff (2004)
Mol. Cancer Ther. 3, 641-646
   Abstract »    Full Text »    PDF »
Dysregulation of the Polo-Like Kinase Pathway in CD4+ T Cells Is Characteristic of Pathogenic Simian Immunodeficiency Virus Infection.
P. Bostik, G. L. Dodd, F. Villinger, A. E. Mayne, and A. A. Ansari (2004)
J. Virol. 78, 1464-1472
   Abstract »    Full Text »    PDF »
Epigenetic Inactivation of CHFR and Sensitivity to Microtubule Inhibitors in Gastric Cancer.
A. Satoh, M. Toyota, F. Itoh, Y. Sasaki, H. Suzuki, K. Ogi, T. Kikuchi, H. Mita, T. Yamashita, T. Kojima, et al. (2003)
Cancer Res. 63, 8606-8613
   Abstract »    Full Text »    PDF »
p190RhoGAP is cell cycle regulated and affects cytokinesis.
L. Su, J. M. Agati, and S. J. Parsons (2003)
J. Cell Biol. 163, 571-582
   Abstract »    Full Text »    PDF »
Events at the end of mitosis in the budding and fission yeasts.
V. Simanis (2003)
J. Cell Sci. 116, 4263-4275
   Abstract »    Full Text »    PDF »
Promotion of Mitosis by Activated Protein Kinase B After DNA Damage Involves Polo-Like Kinase 1 and Checkpoint Protein CHFR.
E. Shtivelman (2003)
Mol. Cancer Res. 1, 959-969
   Abstract »    Full Text »    PDF »
Epigenetic inactivation of CHFR in human tumors.
M. Toyota, Y. Sasaki, A. Satoh, K. Ogi, T. Kikuchi, H. Suzuki, H. Mita, N. Tanaka, F. Itoh, J.-P. J. Issa, et al. (2003)
PNAS 100, 7818-7823
   Abstract »    Full Text »    PDF »
Frequent hypermethylation of the 5' CpG island of the mitotic stress checkpoint gene Chfr in colorectal and non-small cell lung cancer.
P. G. Corn, M. K. Summers, F. Fogt, A. K. Virmani, A. F. Gazdar, T. D. Halazonetis, and W. S. El-Deiry (2003)
Carcinogenesis 24, 47-51
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882