The Cdc14B-Cdh1-Plk1 Axis Controls the G2 DNA-Damage-Response Checkpoint

In response to DNA damage in G2, mammalian cells must avoid entry into mitosis and instead initiate DNA repair. Here, we show that, in response to genotoxic stress in G2, the phosphatase Cdc14B translocates from the nucleolus to the nucleoplasm and induces the activation of the ubiquitin ligase APC/C(Cdh1), with the consequent degradation of Plk1, a prominent mitotic kinase. This process induces the stabilization of Claspin, an activator of the DNA-damage checkpoint, and Wee1, an inhibitor of cell-cycle progression, and allows an efficient G2 checkpoint. As a by-product of APC/C(Cdh1) reactivation in DNA-damaged G2 cells, Claspin, which we show to be an APC/C(Cdh1) substrate in G1, is targeted for degradation. However, this process is counteracted by the deubiquitylating enzyme Usp28 to permit Claspin-mediated activation of Chk1 in response to DNA damage. These findings define a novel pathway that is crucial for the G2 DNA-damage-response checkpoint.

[1]  Hongtao Yu,et al.  A novel motif governs APC-dependent degradation of Drosophila ORC1 in vivo. , 2005, Genes & development.

[2]  J. Peters.  The anaphase promoting complex/cyclosome: a machine designed to destroy , 2006, Nature Reviews Molecular Cell Biology.

[3]  Michele Pagano,et al.  Control of meiotic and mitotic progression by the F box protein beta-Trcp1 in vivo. , 2003, Developmental cell.

[4]  J. Pines,et al.  Ordered proteolysis in anaphase inactivates Plk1 to contribute to proper mitotic exit in human cells , 2004, The Journal of cell biology.

[5]  Michele Pagano,et al.  S6K1- and ßTRCP-Mediated Degradation of PDCD4 Promotes Protein Translation and Cell Growth , 2006, Science.

[6]  J Wade Harper,et al.  The DNA damage response: ten years after. , 2007, Molecular cell.

[7]  Timothy Cardozo,et al.  Control of chromosome stability by the β-TrCP–REST–Mad2 axis , 2008, Nature.

[8]  David O. Morgan,et al.  Sequential E2s Drive Polyubiquitin Chain Assembly on APC Targets , 2007, Cell.

[9]  M. J. van de Vijver,et al.  Gene expression profiling in breast cancer: understanding the molecular basis of histologic grade to improve prognosis. , 2006, Journal of the National Cancer Institute.

[10]  Anastassis Perrakis,et al.  Polo-like Kinase-1 Controls Proteasome-Dependent Degradation of Claspin during Checkpoint Recovery , 2006, Current Biology.

[11]  E. Lees,et al.  The adenovirus E1A-associated kinase consists of cyclin E-p33cdk2 and cyclin A-p33cdk2 , 1993, Journal of virology.

[12]  M. Pagano,et al.  Control of the SCF(Skp2-Cks1) ubiquitin ligase by the APC/C(Cdh1) ubiquitin ligase. , 2004, Nature.

[13]  D. Kletsas,et al.  Evaluation of claspin as a proliferation marker in human cancer and normal tissues , 2007, The Journal of pathology.

[14]  Richard S. Paules,et al.  An ATR- and Chk1-Dependent S Checkpoint Inhibits Replicon Initiation following UVC-Induced DNA Damage , 2002, Molecular and Cellular Biology.

[15]  M. Pagano,et al.  JHDM1B/FBXL10 is a nucleolar protein that represses transcription of ribosomal RNA genes , 2007, Nature.

[16]  Michele Pagano,et al.  Role of the F-Box Protein Skp2 in Adhesion-Dependent Cell Cycle Progression , 2001, The Journal of cell biology.

[17]  S. Meloche,et al.  Phosphorylation of Skp2 regulated by CDK2 and Cdc14B protects it from degradation by APCCdh1 in G1 phase , 2008, The EMBO journal.

[18]  Hiroyuki Osada,et al.  M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Pagano,et al.  APC/C(Cdc20) controls the ubiquitin-mediated degradation of p21 in prometaphase. , 2007, Molecular cell.

[20]  Yi-Song Wang,et al.  The Dual-Specificity Phosphatase CDC14B Bundles and Stabilizes Microtubules , 2005, Molecular and Cellular Biology.

[21]  J. Ruderman,et al.  Identification of a new APC/C recognition domain, the A box, which is required for the Cdh1-dependent destruction of the kinase Aurora-A during mitotic exit. , 2002, Genes & development.

[22]  Michele Pagano,et al.  S6K1- and betaTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. , 2006, Science.

[23]  Shridar Ganesan,et al.  X chromosomal abnormalities in basal-like human breast cancer. , 2006, Cancer cell.

[24]  Michele Pagano,et al.  Control of the SCFSkp2–Cks1 ubiquitin ligase by the APC/CCdh1 ubiquitin ligase , 2004, Nature.

[25]  M. Pagano,et al.  Dual mode of degradation of Cdc25 A phosphatase , 2002, The EMBO journal.

[26]  Todd D. Westergard,et al.  Bimodal Degradation of MLL Protein by the Cell Cycle SCF Skp2 and APC Cdc20 E3 Lilgases Assures Cell Cycle Execution: A Critical Regulatory Circuit Lost in Leukemogenic MLL-Fusions. , 2007 .

[27]  Han Liu,et al.  Bimodal degradation of MLL by SCFSkp2 and APCCdc20 assures cell cycle execution: a critical regulatory circuit lost in leukemogenic MLL fusions. , 2007, Genes & development.

[28]  Angelika Amon,et al.  At the interface between signaling and executing anaphase--Cdc14 and the FEAR network. , 2004, Genes & development.

[29]  Jun Qin,et al.  SCFbeta-TRCP links Chk1 signaling to degradation of the Cdc25A protein phosphatase. , 2003, Genes & development.

[30]  G. Enders,et al.  Dual Regulation of the Anaphase Promoting Complex in Human Cells by Cyclin A-Cdk2 and Cyclin A-Cdk1 Complexes , 2006, Cell cycle.

[31]  Kathleen R. Cho,et al.  Fibroblast growth factor 9 has oncogenic activity and is a downstream target of Wnt signaling in ovarian endometrioid adenocarcinomas. , 2006, Cancer research.

[32]  M. Nakao,et al.  Activation of Cdh1‐dependent APC is required for G1 cell cycle arrest and DNA damage‐induced G2 checkpoint in vertebrate cells , 2001, The EMBO journal.

[33]  R. Medema,et al.  Polo-like kinase-1 is a target of the DNA damage checkpoint , 2000, Nature Cell Biology.

[34]  M. Pagano,et al.  Stabilizers and destabilizers controlling cell cycle oscillators. , 2006, Molecular cell.

[35]  N. Mailand,et al.  Deregulated human Cdc14A phosphatase disrupts centrosome separation and chromosome segregation , 2002, Nature Cell Biology.

[36]  John T. Wei,et al.  Integrative molecular concept modeling of prostate cancer progression , 2007, Nature Genetics.

[37]  A. Fong,et al.  Genetic Evidence for the Essential Role of β-Transducin Repeat-containing Protein in the Inducible Processing of NF-κB2/p100* , 2002, The Journal of Biological Chemistry.

[38]  A. Fong,et al.  Genetic evidence for the essential role of beta-transducin repeat-containing protein in the inducible processing of NF-kappa B2/p100. , 2002, The Journal of biological chemistry.

[39]  F. Bassermann,et al.  NIPA Defines an SCF-Type Mammalian E3 Ligase that Regulates Mitotic Entry , 2005, Cell.

[40]  J. Bartek,et al.  DNA damage checkpoints: from initiation to recovery or adaptation. , 2007, Current opinion in cell biology.

[41]  R. Bernards,et al.  A System for Stable Expression of Short Interfering RNAs in Mammalian Cells , 2002, Science.

[42]  Michele Pagano,et al.  SCFbetaTrCP-mediated degradation of Claspin regulates recovery from the DNA replication checkpoint response. , 2006, Molecular cell.

[43]  S. Elledge,et al.  Human Claspin works with BRCA1 to both positively and negatively regulate cell proliferation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Jiri Bartek,et al.  Cell-cycle checkpoints and cancer , 2004, Nature.

[45]  T. Mak,et al.  A Role for the Deubiquitinating Enzyme USP28 in Control of the DNA-Damage Response , 2006, Cell.

[46]  Jiri Bartek,et al.  Accumulation of cyclin B1 requires E2F and cyclin-A-dependent rearrangement of the anaphase-promoting complex , 1999, Nature.

[47]  D. Botstein,et al.  Gene expression patterns in human liver cancers. , 2002, Molecular biology of the cell.

[48]  Michele Pagano,et al.  SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27 , 1999, Nature Cell Biology.

[49]  Michele Pagano,et al.  Degradation of Cdc25A by β-TrCP during S phase and in response to DNA damage , 2003, Nature.

[50]  Z. Chang,et al.  Mitotic Degradation of Human Thymidine Kinase 1 Is Dependent on the Anaphase-Promoting Complex/Cyclosome-Cdh1-Mediated Pathway , 2004, Molecular and Cellular Biology.

[51]  Jayant P. Menon,et al.  Neuronal and glioma-derived stem cell factor induces angiogenesis within the brain. , 2006, Cancer cell.

[52]  C. Ball,et al.  Identification of genes periodically expressed in the human cell cycle and their expression in tumors. , 2002, Molecular biology of the cell.

[53]  J. Peters,et al.  A Conserved Cyclin-Binding Domain Determines Functional Interplay between Anaphase-Promoting Complex–Cdh1 and Cyclin A-Cdk2 during Cell Cycle Progression , 2001, Molecular and Cellular Biology.

[54]  P. Jackson,et al.  Xenopus Cdc14α/β are localized to the nucleolus and centrosome and are required for embryonic cell division , 2004, BMC Cell Biology.

[55]  Jiri Bartek,et al.  Destruction of Claspin by SCFbetaTrCP restrains Chk1 activation and facilitates recovery from genotoxic stress. , 2006, Molecular cell.

[56]  D. Morgan,et al.  Finishing mitosis, one step at a time , 2007, Nature Reviews Molecular Cell Biology.