Abstract
PBK/TOPK (PDZ-binding kinase, T-LAK-cell-originated protein kinase) is a serine-threonine kinase that is overexpressed in a variety of tumor cells but its role in oncogenesis remains unclear. Here we show, by co-immunoprecipitation experiments and yeast two-hybrid analysis, that PBK/TOPK physically interacts with the tumor suppressor p53 through its DNA-binding (DBD) domain in HCT116 colorectal carcinoma cells that express wild-type p53. PBK also binds to p53 mutants carrying five common point mutations in the DBD domain. The PBK–p53 interaction appears to downmodulate p53 transactivation function as indicated by PBK/TOPK knockdown experiments, which show upregulated expression of the key p53 target gene and cyclin-dependent kinase inhibitor p21 in HCT116 cells, particularly after genotoxic damage from doxorubicin. Furthermore, stable PBK/TOPK knockdown cell lines (derived from HCT116 and MCF-7 cells) showed increased apoptosis, G2/M arrest and slower growth as compared to stable empty vector-transfected control cell lines. Gene microarray studies identified additional p53 target genes involved in apoptosis or cell cycling, which were differentially regulated by PBK knockdown. Together, these data suggest that increased levels of PBK/TOPK may contribute to tumor cell development and progression through suppression of p53 function and consequent reductions in the cell-cycle regulatory proteins such as p21. PBK/TOPK may therefore be a valid target for antineoplastic kinase inhibitors to sensitize tumor cells to chemotherapy-induced apoptosis and growth suppression.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Abe Y, Matsumoto S, Kito K, Ueda N . (2000). Cloning and expression of a novel MAPKK-like protein kinase, lymphokine-activated killer T-cell-originated protein kinase, specifically expressed in the testis and activated lymphoid cells. J Biol Chem 275: 21525–21531.
Abe Y, Takeuchi T, Kagawa-Miki L, Ueda N, Shigemoto K, Yasukawa M et al. (2007). A mitotic kinase TOPK enhances Cdk1/cyclin B1-dependent phosphorylation of PRC1 and promotes cytokinesis. J Mol Biol 370: 231–245.
Ayllón V, O'Connor R . (2007). PBK/TOPK promotes tumour cell proliferation through p38 MAPK activity and regulation of the DNA damage response. Oncogene 26: 3451–3461.
Bartke T, Siegmund D, Peters N, Reichwein M, Henkler F, Scheurich P et al. (2001). p53 upregulates cFLIP, inhibits transcription of NF-κB-regulated genes and induces caspase-8-independent cell death in DLD-1 cells. Oncogene 20: 571–580.
Brooks W, Banerjee S, Crawford DF . (2007). G2E3 is a nucleo-cytoplasmic shuttling protein with DNA damage responsive localization. Exp Cell Res 313: 665–676.
Brooks WS, Helton ES, Banerjee S, Venable M, Johnson L, Crawford DF et al. (2008). G2E3 is a dual function ubiquitin ligase required for early embryonic development. J Biol Chem 283: 22304–22315.
Bunz F, Dutriaux A, Lengauer C, Waldman T, Zhou S, Brown JP et al. (1998). Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 282: 1373–1600.
Cote S, Simard C, Lemieux R . (2002). Regulation of growth-related genes by interleukin-6 in murine myeloma cells. Cytokine 20: 113–120.
Dougherty JD, Garcia ADR, Nakano I, Livingstone M, Norris B, Polakiewicz R et al. (2005). PBK/TOPK, a proliferating neural progenitor-specific mitogen-activated protein kinase kinase. J Neurosci 25: 10773–10785.
el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Vogelstein B, Trent JM et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
Gaudet S, Branton D, Lue RA . (2000). Characterization of PDZ-binding kinase, a mitotic kinase. Proc Natl Acad Sci 97: 5167–5172.
Guo N, Krutzsch HC, Inman JK, Roberts DD . (1997). Thrombospondin 1 and type I repeat peptides of thromboapondin 1 specifically induce apoptosis of endothelial cells. Cancer Res 57: 1735–1742.
Hagn F, Klein C, Demmer O, Marchenko N, Vaseva A, Moll UM et al. (2010). BclxL changes conformation upon binding to wild-type but not mutant p53 DNA binding domain. J Biol Chem 285: 3439–3450.
Harada H, Nakagawa K, Saito M, Kohno S, Nagato S, Furukawa K et al. (2003). Introduction of wild-type p53 enhances thrombospondin-1 expression in human glioma cells. Cancer Lett 191: 109–119.
Herrero-Martin D, Osuna D, Ordonez JL, Sevillano V, Martins AS, Mackintosh C et al. (2009). Stable interference of EWS-FLI1 in an Ewing sarcoma cell line impairs IGF-1/IGF-1R signaling and reveals TOPK as a new target. Br J Cancer 101: 80–90.
Jimenez B, Volpert OV, Crawford SE, Febbraio M, Silverstein RL, Bouck N . (2000). Signals leading to apoptosis-dependent inhibition of neovascularization by thrombospondin-1. Nat Med 6: 41–48.
Kern SE, Kinzler KW, Bruskin A, Bruskin A, Jarosz D, Friedman P et al. (1991). Identification of p53 as a sequence-specific DNA-binding protein. Science 252: 1708–1711.
Kiyono T, Hiraiwa A, Fujita M, Hayashi Y, Akiyama T, Ishibashi M . (1997). Binding of high-risk human papillomavirus E6 oncoproteins to the human homologue of the Drosophila discs large tumor suppressor protein. Proc Natl Acad Sci USA 94: 11612–11616.
Komarova EA, Chernov MV, Franks R, Wang K, Armin G, Zelnick CR et al. (1997). Transgenic mice with p53-responsive lacZ: p53 activity varies dramatically during normal development and determines radiation and drug sensitivity in vivo. EMBO J 16: 1391–1400.
Lee SS, Weiss RS, Javier RT . (1997). Binding of human virus oncoproteins to hDlg/SAP97, a mammalian homolog of the Drosophila discs large tumor suppressor protein. Proc Natl Acad Sci USA 94: 6670–6675.
Li M, Zhou J, Ge Y, Matherly LH, Wu G . (2003). The phosphatase MKP1 is a transcriptional target of p53 involved in cell cycle regulation. J Biol Chem 278: 41059–41068.
Matsumine A, Ogai A, Senda T, Okumura N, Satoh K, Baeg GH et al. (1996). Binding of APC to the human homolog of the Drosophila discs large tumor suppressor protein. Science 272: 1020–1023.
Matsumoto S, Abe Y, Fujibuchi T, Takeuchi T, Kito K, Ueda N et al. (2004). Characterization of a MAPKK-like protein kinase TOPK. Biochem Biophys Res Commun 325: 997–1004.
Müller M, Wilder S, Bannasch D, Israeli D, Lehlbach K, Krammer PH et al. (1998). p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs. J Exp Med 188: 2033–2045.
Nandi A, Tidwell M, Karp J, Rapoport AP . (2004). Protein expression of PDZ-binding kinase is strongly down-regulated during terminal differentiation of HL-60 leukemic cells. Blood Cells Mol Dis 32: 240–245.
Nandi AK, Ford T, Fleksher D, Neuman B, Rapoport AP . (2007). Attenuation of DNA damage checkpoint by PBK, a novel mitotic kinase, involves protein–protein interaction with tumor suppressor p53. Biochem Biophys Res Commun 358: 181–188.
Nandi AK, Rapoport AP . (2006). Expression of PDZ-binding kinase (PBK) is regulated by cell cycle-specific transcription factors E2F and CREB/ATF. Leuk Res 30: 437–447.
Oda E, Ohki R, Murasawa H, Nemoto J, Shibue T, Yamashita T et al. (2000). Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. Science 288: 1053–1058.
Oh SM, Zhu F, Cho YY, Lee KW, Kang BS, Kim HG et al. (2007). T-lymphokine-activated killer cell-originated protein kinase functions as a positive regulator of c-Jun-NH2-kinase 1 signaling and H-Ras-induced cell transformation. Cancer Res 67: 5186–5194.
Park J-H, Lin M-L, Nishidate T, Nakamura Y, Katagiri T . (2006). PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen with an oncogenic activity in breast cancer. Cancer Res 66: 9186–9195.
Patel S, George R, Autore F, Fraternali F, Ladbury JE, Nikolova PV . (2008). Molecular interactions of ASPP1 and ASPP2 with the p53 protein family and the apoptotic promoters PUMA and Bax. Nucleic Acids Res 36: 5139–5151.
Pohl U, Wagenknecht B, Naumann U, Weller M . (1999). p53 enhances BAK and CD95 expression in human malignant glioma cells but does not enhance CD95L-induced apoptosis. Cell Physiol Biochem 9: 29–37.
Rikhof B, Corn PG, El-Deiry WS . (2003). Caspase 10 levels are increased following DNA damage in a p53-dependent manner. Cancer Biol Ther 2: 707–712.
Simons-Evelyn M, Bailey-Dell K, Toretsky JA, Ross DD, Fenton R, Kalvakolanu D et al. (2001). PBK/TOPK is a novel mitotic kinase which is upregulated in Burkitt's lymphoma and other highly proliferative malignant cells. Blood Cells Mol Dis 27: 825–829.
Sot B, Freund SMV, Fersht AR . (2007). Comparative biophysical characterization of p53 with the pro-apoptotic BAK and the anti-apoptotic BCL-xL . J Biol Chem 282: 29193–29200.
Soussi T . (2007). p53 alterations in human cancer: more questions than answers. Oncogene 26: 2145–2156.
Soussi T, Asselain B, Hamroun D, Kato S, Ishioka C, Claustres M et al. (2006). Meta-analysis of the p53 mutation database for mutant p53 biological activity reveals a methodologic bias in mutation detection. Clin Cancer Res 12: 62–69.
Strano S, Dell'Orso S, Di Agostino S, Fotemaggi G, Sacchi A, Blandino G . (2007). Mutant p53: an oncogenic transcription factor. Oncogene 26: 2212–2219.
Tomita Y, Marchenko N, Erster S, Nemajerova A, Dehner A, Klein C et al. (2006). WT p53, but not tumor-derived mutants bind to Bcl2 via the DNA binding domain and induce mitochondrial permeabilization. J Biol Chem 281: 8600–8606.
Wendt J, Radetzki S, Haefen C, Hemmati PG, Güner D, Schulze-Osthoff K et al. (2006). Induction of p21CIP/WAF-1 and G2 arrest by ionizing irradiation impedes caspase-3-mediated apoptosis in human carcinoma cells. Oncogene 25: 972–980.
Zhu F, Zykova TA, Kang BS, Wang Z, Ebeling MC, Abe Y et al. (2007). Bidirectional signals transduced by TOPK–ERK interaction increase tumorigenesis of HCT116 colorectal cancer cells. Gastroenterology 133: 219–231.
Zykova TA, Zhu F, Lu C, Higgins LA, Tatsumi Y, Abe Y et al. (2006). Lymphokine-activated killer T-cell-originated protein kinase phosphorylation of histone H2AX prevents arsenite-induced apoptosis in RPMI17951 melanoma cells. Clin Cancer Res 12: 6884–6893.
Acknowledgements
This work was supported in part by philanthropic gifts from Mr Willard Hackerman, Mr and Mrs Robert Becker, and Mr and Mrs Gaylord Christle. Additional funds were provided by the Marlene and Stewart Greenebaum Cancer Center.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Rights and permissions
About this article
Cite this article
Hu, F., Gartenhaus, R., Eichberg, D. et al. PBK/TOPK interacts with the DBD domain of tumor suppressor p53 and modulates expression of transcriptional targets including p21. Oncogene 29, 5464–5474 (2010). https://doi.org/10.1038/onc.2010.275
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2010.275
Keywords
This article is cited by
-
High expression of PDZ-binding kinase is correlated with poor prognosis and immune infiltrates in hepatocellular carcinoma
World Journal of Surgical Oncology (2022)
-
Colorectal cancer concurrent gene signature based on coherent patterns between genomic and transcriptional alterations
BMC Cancer (2022)
-
PBK drives PARP inhibitor resistance through the TRIM37/NFκB axis in ovarian cancer
Experimental & Molecular Medicine (2022)
-
The design of TOPK inhibitors using structure-based pharmacophore modeling and molecular docking based on an MD-refined homology model
Molecular Diversity (2022)
-
TOPK Activation Exerts Protective Effects on Cisplatin-induced Acute Kidney Injury
Current Medical Science (2022)