Abstract
Casein-kinase CK2 is a Ser/Thr protein kinase that fosters cell survival and proliferation of malignant cells. The CK2 holoenzyme, formed by the association of two catalytic alpha/alpha’ (CK2α/CK2α’) and two regulatory beta subunits (CK2β), phosphorylates diverse intracellular proteins partaking in key cellular processes. A handful of such CK2 substrates have been identified as targets for the substrate-binding anticancer peptide CIGB-300. However, since CK2β also contains a CK2 phosphorylation consensus motif, this peptide may also directly impinge on CK2 enzymatic activity, thus globally modifying the CK2-dependent phosphoproteome. To address such a possibility, firstly, we evaluated the potential interaction of CIGB-300 with CK2 subunits, both in cell-free assays and cellular lysates, as well as its effect on CK2 enzymatic activity. Then, we performed a phosphoproteomic survey focusing on early inhibitory events triggered by CIGB-300 and identified those CK2 substrates significantly inhibited along with disturbed cellular processes. Altogether, we provided here the first evidence for a direct impairment of CK2 enzymatic activity by CIGB-300. Of note, both CK2-mediated inhibitory mechanisms of this anticancer peptide (i.e., substrate- and enzyme-binding mechanism) may run in parallel in tumor cells and help to explain the different anti-neoplastic effects exerted by CIGB-300 in preclinical cancer models.
Similar content being viewed by others
References
Salvi M, Cesaro L, Pinna LA (2010) Variable contribution of protein kinases to the generation of the human phosphoproteome: a global weblogo analysis. Biomol Concepts 1(2):185–195. https://doi.org/10.1515/bmc.2010.013
Ruzzene M (1804) Pinna LA (2010) Addiction to protein kinase CK2: A common denominator of diverse cancer cells? Biochim Biophys Acta (BBA) Proteins Proteom 3:499–504
Barata JT (2011) The impact of PTEN regulation by CK2 on PI3K-dependent signaling and leukemia cell survival. Adv Enzyme Regul 51(1):37–49. https://doi.org/10.1016/j.advenzreg.2010.09.012
Scaglioni PP, Yung TM, Cai LF et al (2006) A CK2-dependent mechanism for degradation of the PML tumor suppressor. Cell 126:269–283
Di Maira G, Salvi M, Arrigoni G et al (2005) Protein kinase CK2 phosphorylates and upregulates Akt/PKB. Cell Death Differ 12:668–677
Channavajhala PL, Seldin DC (2002) Functional interaction of protein kinase CK2 and c-Myc in lymphogenesis. Oncogene 21:5280–5288
Chua MMJ, Ortega CE, Sheikh A et al (2017) CK2 in Cancer: cellular and biochemical mechanisms and potential therapeutic target. Pharmaceuticals 10:18. https://doi.org/10.3390/ph10010018
Pinna LA (2002) Protein kinase CK2: a challenge to canons. J Cell Sci 115:3873–3878
Nunez de Villavicencio-Diaz T, Mazola Y, Perera Negrin Y et al (2015) Predicting CK2 beta-dependent substrates using linear patterns. Biochem Biophys Rep 4:20–27. https://doi.org/10.1016/j.bbrep.2015.08.011
Siddiqui-Jain A, Drygin D, Streiner N et al (2010) CX-4945, an orally bioavailable selective inhibitor of protein kinase CK2, inhibits prosurvival and angiogenic signaling and exhibits antitumor efficacy. Can Res 70(24):10288–10298. https://doi.org/10.1158/0008-5472.CAN-10-1893
Laudet B, Barette C, Dulery V et al (2007) Structure-based design of small peptide inhibitors of protein kinase CK2 subunit interaction. Biochem J 408(3):363–373. https://doi.org/10.1042/BJ20070825
Slaton JW, Unger GM, Sloper DT, Davis AT, Ahmed K (2004) Induction of apoptosis by antisense CK2 in human prostate cancer xenograft model. Mol Cancer Res 2(12):712–721
Marschke RF, Borad MJ, McFarland RW et al (2011) Findings from the phase I clinical trials of CX-4945, an orally available inhibitor of CK2. J Clin Oncol 29:3087. https://doi.org/10.1200/jco.2011.29.15_suppl.3087
Solares AM, Santana A, Baladrón I et al (2009) Safety and preliminary efficacy data of a novel Casein Kinase 2 (CK2) peptide inhibitor administered intralesionally at four dose levels in patients with cervical malignancies. BMC Cancer 9:146
Perea SE, Reyes O, Puchades Y et al (2004) Antitumor effect of a novel proapoptotic peptide that impairs the phosphorylation by the protein kinase 2 (casein kinase 2). Can Res 64(19):7127–7129. https://doi.org/10.1158/0008-5472.CAN-04-2086
Perera Y, Farina HG, Gil J et al (2009) Anticancer peptide CIGB-300 binds to nucleophosmin/B23, impairs its CK2-mediated phosphorylation, and leads to apoptosis through its nucleolar disassembly activity. Mol Cancer Ther 8(5):1189–1196. https://doi.org/10.1158/1535-7163.MCT-08-1056
Perea SE, Baladron I, Garcia Y et al (2011) CIGB-300, a synthetic peptide-based drug that targets the CK2 phosphoaceptor domain. Translational and clinical research. Mol Cell Biochem 356:45–50. https://doi.org/10.1007/s11010-011-0950-y
Martins LR, Perera Y, Lucio P et al (2014) Targeting chronic lymphocytic leukemia using CIGB-300, a clinical-stage CK2-specific cell-permeable peptide inhibitor. Oncotarget 5(1):258–263. https://doi.org/10.18632/oncotarget.1513
Boldyreff B, James P, Staudenmann W, Issinger OG (1993) Ser2 is the autophosphorylation site in the beta subunit from bicistronically expressed human casein kinase-2 and from native rat liver casein kinase-2 beta. Eur J Biochem 218(2):515–521. https://doi.org/10.1111/j.1432-1033.1993.tb18404.x
Litchfield DW, Lozeman FJ, Cicirelli MF et al (1991) Phosphorylation of the beta subunit of casein kinase II in human A431 cells. Identification of the autophosphorylation site and a site phosphorylated by p34cdc2. J Biol Chem 266(30):20380–20389
Pagano MA, Sarno S, Poletto G et al (2005) Autophosphorylation at the regulatory beta subunit reflects the supramolecular organization of protein kinase CK2. Mol Cell Biochem 274:23–29. https://doi.org/10.1007/s11010-005-3116-y
Zhang C, Vilk G, Canton DA, Litchfield DW (2002) Phosphorylation regulates the stability of the regulatory CK2beta subunit. Oncogene 21(23):3754–3764. https://doi.org/10.1038/sj.onc.1205467
Rodriguez-Ulloa A, Ramos Y, Gil J et al (2010) Proteomic profile regulated by the anticancer peptide CIGB-300 in non-small cell lung cancer (NSCLC) cells. J Proteome Res 9(10):5473–5483. https://doi.org/10.1021/pr100728v
Leroy D, Filhol O, Quintaine N et al (1999) Dissecting subdomains involved in multiple functions of the CK2beta subunit. Mol Cell Biochem 191:43–50
Huang H, Arighi CN, Ross KE et al (2018) iPTMnet: an integrated resource for protein post-translational modification network discovery. Nucleic Acids Res 46:D542–D550. https://doi.org/10.1093/nar/gkx1104
Crooks GE, Hon G, Chandonia JM, Brenner SE (2004) WebLogo: a sequence logo generator. Genome Res 14(6):1188–1190. https://doi.org/10.1101/gr.849004
Lachmann A, Ma’ayan A (2009) KEA: kinase enrichment analysis. Bioinformatics 25(5):684–686. https://doi.org/10.1093/bioinformatics/btp026
Weidner C, Fischer C, Sauer S (2014) PHOXTRACK-a tool for interpreting comprehensive datasets of post-translational modifications of proteins. Bioinformatics 30(23):3410–3411. https://doi.org/10.1093/bioinformatics/btu572
Kuleshov MV, Jones MR, Rouillard AD et al (2016) Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic Acids Res 44(W1):W90–97. https://doi.org/10.1093/nar/gkw377
Romero-Oliva F, Jacob G, Allende JE (2003) Dual effect of lysine-rich polypeptides on the activity of protein kinase CK2. Cell Biochem 89(2):348–355
Szebeni A, Hingorani K, Negi S, Olson MOJ (2003) Role of protein kinase CK2 phosphorylation in the molecular chaperone activity of nucleolar protein b23. J Biol Chem 278(11):9107–9115. https://doi.org/10.1074/jbc.M204411200
Adenuga D, Rahman I (2010) Protein kinase CK2-mediated phosphorylation of HDAC2 regulates co-repressor formation, deacetylase activity and acetylation of HDAC2 by cigarette smoke and aldehydes. Arch Biochem Biophys 498(1):62–73. https://doi.org/10.1016/j.abb.2010.04.002
Zanin S, Sandre M, Cozza G et al (2015) Chimeric peptides as modulators of CK2-dependent signaling: Mechanism of action and off-target effects. Biochem Biophys Acta 1854:1694–1707. https://doi.org/10.1016/j.bbapap.2015.04.026
Paytubi S, Wang X, Lam YW et al (2009) ABC50 promotes translation initiation in mammalian cells. J Biol Chem 284(36):24061–24073. https://doi.org/10.1074/jbc.M109.031625
Salvi M, Xu D, Chen Y (2009) Programmed cell death protein 5 (PDCD5) is phosphorylated by CK2 in vitro and in 293T cells. Biochem Biophys Res Commun 387(3):606–610. https://doi.org/10.1016/j.bbrc.2009.07.067
Li G, Ma D (1863) Chen Y (2016) Cellular functions of programmed cell death 5. Biochem Biophys Acta 4:572–580. https://doi.org/10.1016/j.bbamcr.2015.12.021
Miyata Y, Nishida E (2004) CK2 controls multiple protein kinases by phosphorylating a kinase-targeting molecular chaperone, Cdc37. Mol Cell Biol 24(9):4065–4074. https://doi.org/10.1128/mcb.24.9.4065-4074.2004
Kim SW, Hasanuzzaman M, Cho M et al (2015) Casein Kinase 2 (CK2)-mediated phosphorylation of Hsp90beta as a novel mechanism of rifampin-induced MDR1 expression. J Biol Chem 290(27):17029–17040. https://doi.org/10.1074/jbc.M114.624106
Betapudi V, Gokulrangan G, Chance MR, Egelhoff TT (2011) A proteomic study of myosin II motor proteins during tumor cell migration. J Mol Biol 407(5):673–686. https://doi.org/10.1016/j.jmb.2011.02.010
Yu W, Ding X, Chen F et al (2009) The phosphorylation of SEPT2 on Ser218 by casein kinase 2 is important to hepatoma carcinoma cell proliferation. Mol Cell Biochem 325:61–67. https://doi.org/10.1007/s11010-008-0020-2
Khoronenkova SV, Dianova II, Ternette N et al (2012) ATM-dependent downregulation of USP7/HAUSP by PPM1G activates p53 response to DNA damage. Mol Cell 45(6):801–813. https://doi.org/10.1016/j.molcel.2012.01.021
Moreno FJ, Avila J (1998) Phosphorylation of stathmin modulates its function as a microtubule depolymerizing factor. Mol Cell Biochem 183:201–209. https://doi.org/10.1023/a:1006807814580
Polzien L, Baljuls A, Rennefahrt UEE et al (2009) Identification of novel in vivo phosphorylation sites of the human proapoptotic protein BAD: pore-forming activity of BAD is regulated by phosphorylation. J Biol Chem 284(41):28004–28020. https://doi.org/10.1074/jbc.M109.010702
Bui NLC, Pandey V, Zhu T, Ma L, Basappa Lobie PE (2018) Bad phosphorylation as a target of inhibition in oncology. Cancer Lett 415:177–186. https://doi.org/10.1016/j.canlet.2017.11.017
St-Denis N, Gabriel M, Turowec JP et al (2015) Systematic investigation of hierarchical phosphorylation by protein kinase CK2. J Proteom 118:49–62. https://doi.org/10.1016/j.jprot.2014.10.020
Rusin SF, Adamo ME, Kettenbach AN (2017) Identification of candidate casein kinase 2 substrates in mitosis by quantitative phosphoproteomics. Front Cell Dev Biol 5:97. https://doi.org/10.3389/fcell.2017.00097
Franchin C, Cesaro L, Salvi M et al (1854) (2015) Quantitative analysis of a phosphoproteome readily altered by the protein kinase CK2 inhibitor quinalizarin in HEK-293T cells. Biochim Biophys Acta 6:609–623. https://doi.org/10.1016/j.bbapap.2014.09.017
Salvi M, Sarno S, Cesaro L, Nakamura H, Pinna LA (2009) Extraordinary pleiotropy of protein kinase CK2 revealed by weblogo phosphoproteome analysis. Biochem Biophys Acta 1793(5):847–859. https://doi.org/10.1016/j.bbamcr.2009.01.013
Franchin C, Borgo C, Zaramella S et al (2017) Exploring the CK2 paradox: restless, dangerous, dispensable. Pharmaceuticals 10(1):11. https://doi.org/10.3390/ph10010011
Funding
This work was conducted with the financial support of the CIGB-300 Grant, Biomedical Research Division, CIGB, Cuba.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Perera, Y., Ramos, Y., Padrón, G. et al. CIGB-300 anticancer peptide regulates the protein kinase CK2-dependent phosphoproteome. Mol Cell Biochem 470, 63–75 (2020). https://doi.org/10.1007/s11010-020-03747-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-020-03747-1