Cloning and Functional Characterization of Ptpcd2 as a Novel Cell Cycle Related Protein Tyrosine Phosphatase that Regulates Mitotic Exit

  • Zineldeen, Doaa H. (Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Tanta University) ;
  • Wagih, Ayman A. (Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Tanta University) ;
  • Nakanishi, Makoto (Department of Cell Biology, Graduate School of Medical Sciences, Nagoya City University)
  • Published : 2013.06.30


Faithful transmission of genetic information depends on accurate chromosome segregation as cells exit from mitosis, and errors in chromosomal segregation are catastrophic and may lead to aneuploidy which is the hallmark of cancer. In eukaryotes, an elaborate molecular control system ensures proper orchestration of events at mitotic exit. Phosphorylation of specific tyrosyl residues is a major control mechanism for cellular proliferation and the activities of protein tyrosine kinases and phosphatases must be integrated. Although mitotic kinases are well characterized, phosphatases involved in mitosis remain largely elusive. Here we identify a novel variant of mouse protein tyrosine phosphatase containing domain 1 (Ptpcd1), that we named Ptpcd2. Ptpcd1 is a Cdc14 related centrosomal phosphatase. Our newly identified Ptpcd2 shared a significant homology to yeast Cdc14p (34.1%) and other Cdc14 family of phosphatases. By subcellular fractionation Ptpcd2 was found to be enriched in the cytoplasm and nuclear pellets with catalytic phosphatase activity. By means of immunofluorescence, Ptpcd2 was spatiotemporally regulated in a cell cycle dependent manner with cytoplasmic abundance during mitosis, followed by nuclear localization during interphase. Overexpression of Ptpcd2 induced mitotic exit with decreased levels of some mitotic markers. Moreover, Ptpcd2 failed to colocalize with the centrosomal marker ${\gamma}$-tubulin, suggesting it as a non-centrosomal protein. Taken together, Ptpcd2 phosphatase appears a non-centrosomal variant of Ptpcd1 with probable mitotic functions. The identification of this new phosphatase suggests the existence of an interacting phosphatase network that controls mammalian mitosis and provides new drug targets for anticancer modalities.


Ptpcd1;mitotic exit;cell cycle;Cdc14;protein tyrosine phosphatase


  1. Bas a tumor suppressor in hepatocellular carcinogenesis. Cancer Cell, 19, 629-39.
  2. Berdougo E, Nachury MV, Jackson PK, Jallepalli PV (2008). The nucleolar phosphatase Cdc14B is dispensable for chromosome segregation and mitotic exit in human cells. Cell Cycle, 7, 1184-90.
  3. Beuming T, Skrabanek L, Niv MY, Mukherjee P, Weinstein H (2005). PDZBase: a protein-protein interaction database for PDZ-domains. Bioinformatics, 21, 827-28.
  4. Bremmer SC, Hall H, Martinez JS, et al (2012). Cdc14 phosphatases preferentially dephosphorylate a subset of cyclin-dependent kinase (Cdk) sites containing phosphoserine. J Biol Chem, 287, 1662-9.
  5. Chan KS, Koh CG, Li HY (2012). Mitosis-targeted anti-cancer therapies: where they stand. Cell Death Dis, 3, 411.
  6. Chen J, Chan AW, To KF, et al (2013). SIRT2 overexpression in hepatocellular carcinoma mediates epithelial to mesenchymal transition by protein kinase B/glycogen synthase kinase-$3\beta$/$\beta $-catenin signaling. Hepatology, 57, 2287-98.
  7. Chiesa M, Guillamot M, Bueno MJ, Malumbres M (2011). The Cdc14B phosphatase displays oncogenic activity mediated by the Ras-Mek signaling pathway. Cell Cycle, 10, 1607-17.
  8. Cho HP, Liu Y, Gomez M, et al (2005). The dual-specificity phosphatase CDC14B bundles and stabilizes microtubules. Mol Cell Biol, 25, 4541-51.
  9. Dinkel H, Michael S, Weatheritt RJ, et al (2012). ELM--the database of eukaryotic linear motifs. Nucleic Acids Res, 40, 242-51.
  10. Dryden SC, Nahhas FA, Nowak JE, Goustin AS, Tainsky MA (2003). Role for human SIRT2 NAD-dependent deacetylase activity in control of mitotic exit in the cell cycle. Mol Cell Biol, 23, 3173-85.
  11. Galeano F, Rossetti C, Tomaselli S, et al (2013). ADAR2-editing activity inhibits glioblastoma growth through the modulation of the CDC14B/Skp2/p21/p27 axis. Oncogene, 32, 998-1009.
  12. Galan-Malo P, Vela L, Gonzalo O, et al (2012). Cell fate after mitotic arrest in different tumor cells is determined by the balance between slippage and apoptotic threshold. Toxicol Appl Pharmacol, 258, 384-93.
  13. Gascoigne KE, Cheeseman IM (2013). CDK-dependent phosphorylation and nuclear exclusion coordinately control kinetochore assembly state. J Cell Biol, 201, 23-32.
  14. Gascoigne KE, Taylor SS (2008). Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell, 14, 111-22.
  15. Graham JM (2002). Preparation of crude subcellular fractions by differential centrifugation. Sci World J, 2, 1638-42.
  16. Hancioglu B, Tyson JJ (2012). A mathematical model of mitotic exit in budding yeast: the role of Polo kinase. PLoS One, 7, 308-10.
  17. Huang HC, Shi J, Orth JD, Mitchison TJ (2009). Evidence that mitotic exit is a better cancer therapeutic target than spindle assembly. Cancer Cell, 16, 347-58.
  18. Hunt T (2013). On the regulation of protein phosphatase 2A and its role in controlling entry into and exit from mitosis. Adv Biol Regul, 53, 173-8.
  19. Janssen A, Medema RH (2011). Mitosis as an anti-cancer target. Oncogene, 30, 2799-809.
  20. Lai CC, Lin PM, Lin SF, et al (2013). Altered expression of SIRT gene family in head and neck squamous cell carcinoma. Tumour Biol, 34, 1847-54.
  21. Larkin MA, Blackshields G, Brown NP, et al. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947-48.
  22. Lupas A, Van Dyke M, Stock, J. (1991). Predicting coiled coils from protein sequences. Science, 252, 1162-64.
  23. Manchado E, Guillamot M, de Carcer G, et al (2010). Targeting mitotic exit leads to tumor regression in vivo: Modulation by Cdk1, Mastl, and the PP2A/$B55\alpha$, $\delta$ phosphatase. Cancer Cell, 18, 641-54.
  24. Massey AJ, Borgognoni J, Bentley C (2010). Context-dependent cell cycle checkpoint abrogation by a novel kinase inhibitor. PLoS One, 5, 13123.
  25. Maya-Mendoza A, Jackson, DA (2009). Replication timing: findings from fibers. Cell Cycle, 8, 3073-74.
  26. Mocciaro A, Berdougo E, Zeng K (2010). Vertebrate cells genetically deficient for Cdc14A or Cdc14B retain DNA damage checkpoint proficiency but are impaired in DNA repair. J Cell Biol, 189, 631-9.
  27. Mocciaro A, Schiebel E. (2010). Cdc14: a highly conserved family of phosphatases with non-conserved functions? J Cell Sci, 123, 2867-76.
  28. Mochida S, Hunt T (2007). Calcineurin is required to release Xenopus egg extracts from meiotic M phase. Nature, 449, 336-40.
  29. Mendez J, Stillman B (2000). Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis. Mol Cell Biol, 20, 8602-12.
  30. Nalepa G, Harper JW (2004). Visualization of a highly organized intranuclear network of filaments in living mammalian cells. Cell Motil Cytoskeleton, 59, 94-108.
  31. Naruyama H, Shimada M, Niida H, et al (2008). Essential role of Chk1 in S phase progression through regulation of RNR2 expression. Biochem Biophys Res Commun, 374, 79-83.
  32. Niida H, Katsuno Y, Banerjee B, Hande MP, Nakanishi M (2007). Specific role of Chk1 phosphorylations in cell survival and checkpoint activation. Mol Cell Biol, 27, 2572-81.
  33. North BJ, Verdin E (2007). Mitotic regulation of SIRT2 by cyclin-dependent kinase 1-dependent phosphorylation. J Biol Chem, 282, 19546-55.
  34. Novak B, Tyson JJ, Gyorffy B, Csikasz-Nagy A (2007). Irreversible cell-cycle transitions are due to systems-level feedback. Nat Cell Biol, 9, 724-28.
  35. Patterson KI, Brummer T, O'Brien PM, Daly RJ (2009). Dualspecificity phosphatases: critical regulators with diverse cellular targets. Biochem J, 418, 475-89.
  36. Rossio V, Galati E, Ferrari M, et al (2010). The RSC chromatinremodeling complex influences mitotic exit and adaptation to the spindle assembly checkpoint by controlling the Cdc14 phosphatase. J Cell Biol, 191, 981-97.
  37. Russell P, Hennessy BT, Li J, et al (2012). Cyclin G1 regulates the outcome of taxane-induced mitotic checkpoint arrest. Oncogene, 31, 2450-60.
  38. Sanchez-Diaz A, Nkosi PJ, Murray S, Labib K (2012). The Mitotic Exit Network and Cdc14 phosphatase initiate cytokinesis by counteracting CDK phosphorylations and blocking polarised growth. EMBO J, 31, 3620-34.
  39. Shimada M, Nakanishi M (2013). Response to DNA damage: why do we need to focus on protein phosphatases? Front Oncol, 3, 8.
  40. Shimada M, Niida H, Zineldeen DH, et al (2008). Chk1 is a histone H3 threonine 11 kinase that regulates DNA damageinduced transcriptional repression. Cell, 132, 221-32.
  41. Skoufias DA, Indorato RL, Lacroix F, Panopoulos A, Margolis RL (2007). Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed. J Cell Biol, 179, 671-85.
  42. Takeo S, Hawley RS, Aigaki T (2010). Calcineurin and its regulation by Sra/RCAN is required for completion of meiosis in Drosophila. Dev Biol, 344, 957-67.
  43. Theobald B, Bonness K, Musiyenko A (2013). Suppression of ser/thr phosphatase 4 (PP4C/PPP4C) mimics a novel post-mitotic action of fostriecin, producing mitotic slippage followed by tetraploid cell death. Mol Cancer Res, [Epub ahead of print].
  44. Uchida T, Matozaki T, Matsuda K (1993). Phorbol ester stimulates the activity of a protein tyrosine phosphatase containing SH2 domains (PTP1C) in HL-60 leukemia cells by increasing gene expression. J Biol Chem, 268, 11845-50.
  45. van den Berk LC, Landi E, Harmsen E, Dente L, Hendriks WJ (2005). Redox-regulated affinity of the third PDZ domain in the phosphotyrosine phosphatase PTP-BL for cysteinecontaining target peptides. FEBS J, 272, 3306-16.
  46. Visconti R, Palazzo L, Della Monica R, Grieco D (2012). Fcp1-dependent dephosphorylation is required for M-phasepromoting factor inactivation at mitosis exit. Nat Commun, 3, 894.
  47. Vazquez-Novelle MD, Esteban V, Bueno A, Sacristan MP. (2005). Functional homology among human and fission yeast Cdc14 phosphatases. J Biol Chem, 280, 29144-50.
  48. Wei Z, Zhang P (2011). A phosphatase turns aggressive: the oncogenicity of Cdc14B. Cell Cycle, 10, 2414.
  49. Wu J, Cho HP, Rhee DB, et al (2008). Cdc14B depletion leads to centriole amplification, and its overexpression prevents unscheduled centriole duplication. J Cell Biol, 181, 475-83.
  50. Wurzenberger C, Gerlich DW (2011). Phosphatases: providing safe passage through mitotic exit. Nat Rev Mol Cell Biol, 12, 469-82.
  51. Zhang Q, Claret FX (2012). Phosphatases: the new brakes for cancer development? Enzyme Res, 2012, 659649.
  52. Zineldeen DH, Shimada M, Niida H, Katsuno Y, Nakanishi M (2009). Ptpcd-1 is a novel cell cycle related phosphatase that regulates centriole duplication and cytokinesis. Biochem Biophys Res Commun, 380, 460-6.

Cited by

  1. Alternative Chk1-independent S/M checkpoint in somatic cells that prevents premature mitotic entry vol.34, pp.4, 2017,