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Isoform specific phosphorylation of p53 by protein kinase CK1

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An Erratum to this article was published on 05 January 2011

Abstract

The ability of three isoforms of protein kinase CK1 (α, γ1, and δ) to phosphorylate the N-terminal region of p53 has been assessed using either recombinant p53 or a synthetic peptide reproducing its 1–28 sequence. Both substrates are readily phosphoylated by CK1δ and CK1α, but not by the γ isoform. Affinity of full size p53 for CK1 is 3 orders of magnitude higher than that of its N-terminal peptide (K m 0.82 μM vs 1.51 mM). The preferred target is S20, whose phosphorylation critically relies on E17, while S6 is unaffected despite displaying the same consensus (E-x-x-S). Our data support the concept that non-primed phosphorylation of p53 by CK1 is an isoform-specific reaction preferentially affecting S20 by a mechanism which is grounded both on a local consensus and on a remote docking site mapped to the K221RQK224 loop according to modeling and mutational analysis.

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References

  1. Knippschild U, Gocht A, Wolff S, Huber N, Löhler J, Stöter M (2005) The casein kinase 1 family: participation in multiple cellular processes in eukaryotes. Cell Signal 17(6):675–689

    Article  CAS  PubMed  Google Scholar 

  2. Longenecker KL, Roach PJ, Hurley TD (1996) Three-dimensional structure of mammalian casein kinase I: molecular basis for phosphate recognition. J Mol Biol 257:618–631

    Article  CAS  PubMed  Google Scholar 

  3. Xu RM, Carmel G, Sweet RM, Kuret J, Cheng X (1995) Crystal structure of casein kinase-1, a phosphate-directed protein kinase. EMBO J 14:1015–1023

    CAS  PubMed  Google Scholar 

  4. Brockman JL, Gross SD, Sussman MR, Anderson RA (1992) Cell cycle-dependent localization of casein kinase I to mitotic spindles. Proc Natl Acad Sci USA 89:9454–9458

    Article  CAS  PubMed  Google Scholar 

  5. Milne DM, Looby P, Meek DW (2001) Catalytic activity of protein kinase CK1 delta (casein kinase 1 delta) is essential for its normal subcellular localization. Exp Cell Res 263:43–54

    Article  CAS  PubMed  Google Scholar 

  6. Petronczki M, Matos J, Mori S, Gregan J, Bogdanova A, Schwickart M, Mechtler K, Shirahige K, Zachariae W, Nasmyth K (2006) Monopolar attachment of sister kinetochores at meiosis I requires casein kinase 1. Cell 126:1049–1064

    Article  CAS  PubMed  Google Scholar 

  7. Behrend L, Milne DM, Stoter M, Deppert W, Campbell LE, Meek DW, Knippschild U (2000) IC261, a specific inhibitor of the protein kinases casein kinase 1-delta and -epsilon, triggers the mitotic checkpoint and induces p53-dependent postmitotic effects. Oncogene 19:5303–5313

    Article  CAS  PubMed  Google Scholar 

  8. Behrend L, Stoter M, Kurth M, Rutter G, Heukeshoven J, Deppert W, Knippschild U (2000) Interaction of casein kinase 1 delta (CK1delta) with post-Golgi structures, microtubules and the spindle apparatus. Eur J Cell Biol 79:240–251

    Article  CAS  PubMed  Google Scholar 

  9. Camacho F, Cilio M, Guo Y, Virshup DM, Patel K, Khorkova O, Styren S, Morse B, Yao Z, Keesler GA (2001) Human casein kinase I delta phosphorylation of human circadian clock proteins period 1 and 2. FEBS Lett 489:159–165

    Article  CAS  PubMed  Google Scholar 

  10. Zhu J, Shibasaki F, Price R, Guillemot JC, Yano T, Dotsch V, Wagner G, Ferrara P, McKeon F (1998) Intramolecular masking of nuclear import signal on NF-AT4 by casein kinase I and MEKK1. Cell 93:851–861

    Article  CAS  PubMed  Google Scholar 

  11. Peters JM, McKay RM, McKay JP, Graff JM (1999) Casein kinase I transduces Wnt signals. Nature 401:345–350

    Article  CAS  PubMed  Google Scholar 

  12. Zeng X, Tamai K, Doble B, Li S, Huang H, Habas R, Okamura H, Woodgett J, He X (2005) A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. Nature 438:873–877

    Article  CAS  PubMed  Google Scholar 

  13. Davidson G, Wu W, Shen J, Bilic J, Fenger U, Stannek P, Glinka A, Niehrs C (2005) Casein kinase 1 gamma couples Wnt receptor activation to cytoplasmic signal transduction. Nature 438:867–872

    Article  CAS  PubMed  Google Scholar 

  14. Hämmerlein A, Weiske J, Huber O (2005) A second protein kinase CK1-mediated step negatively regulates Wnt signalling by disrupting the lymphocyte enhancer factor-1/beta-catenin complex. Cell Mol Life Sci 62(5):606–618

    Article  PubMed  Google Scholar 

  15. Swiatek W, Kang H, Garcia BA, Shabanowitz J, Coombs GS, Hunt DF, Virshup DM (2006) Negative regulation of LRP6 function by casein kinase I epsilon phosphorylation. J Biol Chem 281:12233–12241

    Article  CAS  PubMed  Google Scholar 

  16. Price MA (2006) CKI, there’s more than one: casein kinase I family members in Wnt and Hedgehog signaling. Genes Dev 20(4):399–410

    Article  CAS  PubMed  Google Scholar 

  17. Bryja V, Schulte G, Rawal N, Grahn A, Arenas E (2007) Wnt-5a induces Dishevelled phosphorylation and dopaminergic differentiation via a CK1-dependent mechanism. J Cell Sci 120(4):586–595

    Article  CAS  PubMed  Google Scholar 

  18. Beyaert R, Vanhaesebroeck B, Declercq W, Van Lint J, Vandenabele P, Agostinis P, Vandenheede JR, Fiers W (1995) Casein kinase-1 phosphorylates the p75 tumor necrosis factor receptor and negatively regulates tumor necrosis factor signaling for apoptosis. J Biol Chem 270:23293–23299

    Article  CAS  PubMed  Google Scholar 

  19. Desagher S, Osen-Sand A, Montessuit S, Magnenat E, Vilbois F, Hochmann A, Journot L, Antonsson B, Martinou JC (2001) Phosphorylation of bid by casein kinases I and II regulates its cleavage by caspase 8. Mol Cell 8:601–611

    Article  CAS  PubMed  Google Scholar 

  20. Schwab C, DeMaggio AJ, Ghoshal N, Binder LI, Kuret J, McGeer PL (2000) Casein kinase 1 delta is associated with pathological accumulation of tau in several neurodegenerative diseases. Neurobiol Aging 21:503–510

    Article  CAS  PubMed  Google Scholar 

  21. Yasojima K, Kuret J, DeMaggio AJ, McGeer E, McGeer PL (2000) Casein kinase 1 delta mRNA is upregulated in Alzheimer disease brain. Brain Res 865:116–120

    Article  CAS  PubMed  Google Scholar 

  22. Xu Y, Padiath QS, Shapiro RE, Jones CR, Wu SC, Saigoh N, Saigoh K, Ptacek LJ, Fu YH (2005) Functional consequences of a CKI delta mutation causing familial advanced sleep phase syndrome. Nature 434:640–644

    Article  CAS  PubMed  Google Scholar 

  23. Elias L, Li AP, Longmire J (1981) Cyclic adenosine 3′:5′-monophosphate-dependent and -independent protein kinase in acute myeloblastic leukemia. Cancer Res 41:2182–2188

    CAS  PubMed  Google Scholar 

  24. Mishra SK, Yang Z, Mazumdar A, Talukder AH, Larose L, Kumar R (2004) Metastatic tumor antigen 1 short form (MTA1 s) associates with casein kinase I-gamma2, an estrogen-responsive kinase. Oncogene 23:4422–4429

    Article  CAS  PubMed  Google Scholar 

  25. Frierson HF Jr, El-Naggar AK, Welsh JB, Sapinoso LM, Su AI, Cheng J, Saku T, Moskaluk CA, Hampton GM (2002) Large scale molecular analysis identifies genes with altered expression in salivary adenoid cystic carcinoma. Am J Pathol 161:1315–1323

    CAS  PubMed  Google Scholar 

  26. Fuja TJ, Lin F, Osann KE, Bryant PJ (2004) Somatic mutations and altered expression of the candidate tumor suppressors CSNK1 epsilon, DLG1, and EDD/hHYD in mammary ductal carcinoma. Cancer Res 64:942–951

    Article  CAS  PubMed  Google Scholar 

  27. Adorno M, Cordenonsi M, Montagner M, Dupont S, Wong C, Hann B, Solari A, Bobisse S, Rondina MB, Guzzardo V, Parenti AR, Rosato A, Bicciato S, Balmain A, Piccolo S (2009) A Mutant-p53/Smad complex opposes p63 to empower TGFbeta-induced metastasis. Cell 137:87–98

    Article  CAS  PubMed  Google Scholar 

  28. Meggio F, Donella-Deana A, Pinna LA (1979) Studies on the structural requirements of a microsomal cAMP-independent protein kinase. FEBS Lett 106(1):76–80

    Article  CAS  PubMed  Google Scholar 

  29. Flotow H, Graves PR, Wang AQ, Fiol CJ, Roeske RW, Roach PJ (1990) Phosphate groups as substrate determinants for casein kinase I action. J Biol Chem 265(24):14264–14269

    CAS  PubMed  Google Scholar 

  30. Meggio F, Perich JW, Reynolds EC, Pinna LA (1991) A synthetic beta-casein phosphopeptide and analogues as model substrates for casein kinase-1, a ubiquitous, phosphate directed protein kinase. FEBS Lett 283(2):303–306

    Article  CAS  PubMed  Google Scholar 

  31. Roach PJ (1991) Multisite and hierarchal protein phosphorylation. J Biol Chem 266(22):14139–14142

    CAS  PubMed  Google Scholar 

  32. Amit S, Hatzubai A, Birman Y, Andersen JS, Ben-Shushan E, Mann M, Ben-Neriah Y, Alkalay I (2002) Axin mediated CKI phosphorylation of β-catenin at Ser 45: a molecular switch for the Wnt pathway. Genes Dev 16:1066–1076

    Article  CAS  PubMed  Google Scholar 

  33. Liu C, Li Y, Semenov M, Han C, Baeg GH, Tan Y, Zhang Z, Lin X, He X (2002) Control of β-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell 108:837–847

    Article  CAS  PubMed  Google Scholar 

  34. Ha NC, Tonozuka T, Stamos JL, Choi HJ, Weis WI (2004) Mechanism of phosphorylation-dependent binding of APC to β-catenin and its role in β-catenin degradation. Mol Cell 15:511–521

    Article  CAS  PubMed  Google Scholar 

  35. Xing Y, Clements WK, Le Trong I, Hinds TR, Stenkamp R, Kimelman D, Xu W (2004) Crystal structure of a β-catenin/APC complex reveals a critical role for APC phosphorylation in APC function. Mol Cell 15:523–533

    Article  CAS  PubMed  Google Scholar 

  36. Ferrarese A, Marin O, Bustos VH, Venerando A, Antonelli M, Allende JE, Pinna LA (2007) Chemical dissection of the APC Repeat 3 multistep phosphorylation by the concerted action of protein kinases CK1 and GSK3. Biochemistry 46(42):11902–11910

    Article  CAS  PubMed  Google Scholar 

  37. Desdouits F, Siciliano JC, Greengard P, Girault JA (1995) Dopamine- and cAMP-regulated phosphoprotein DARPP-32: phosphorylation of Ser-137 by casein kinase I inhibits dephosphorylation of Thr-34 by calcineurin. Proc Natl Acad Sci USA 92(7):2682–2685

    Article  CAS  PubMed  Google Scholar 

  38. Pulgar V, Marin O, Meggio F, Allende CC, Allende JE, Pinna LA (1999) Optimal sequences for non-phosphate-directed phosphorylation by protein kinase CK1 (casein kinase-1)–a re-evaluation. Eur J Biochem 260(2):520–526

    Article  CAS  PubMed  Google Scholar 

  39. Marin O, Burzio V, Boschetti M, Meggio F, Allende CC, Allende JE, Pinna LA (2002) Structural features underlying the multisite phosphorylation of the A domain of the NF-AT4 transcription factor by protein kinase CK1. Biochemistry 41(2):618–627

    Article  CAS  PubMed  Google Scholar 

  40. Marin O, Bustos VH, Cesaro L, Meggio F, Pagano MA, Antonelli M, Allende CC, Pinna LA, Allende JE (2003) A noncanonical sequence phosphorylated by casein kinase 1 in beta-catenin may play a role in casein kinase 1 targeting of important signaling proteins. Proc Natl Acad Sci USA 100(18):10193–10200

    Article  CAS  PubMed  Google Scholar 

  41. Bustos VH, Ferrarese A, Venerando A, Marin O, Allende JE, Pinna LA (2006) The first armadillo repeat is involved in the recognition and regulation of beta-catenin phosphorylation by protein kinase CK1. Proc Natl Acad Sci USA 103(52):19725–19730

    Article  CAS  PubMed  Google Scholar 

  42. Bode AM, Dong Z (2004) Post-translational modification of p53 in tumorigenesis. Nat Rev Cancer 4:793–805

    Article  CAS  PubMed  Google Scholar 

  43. Lacroix M, Toillon RA, Leclercq G (2006) p53 and breast cancer, an update. Endocr Relat Cancer 13(2):293–325

    Article  CAS  PubMed  Google Scholar 

  44. Appella E, Anderson CW (2001) Post-translational modifications and activation of p53 by genotoxic stresses. Eur J Biochem 268(10):2764–2772

    Article  CAS  PubMed  Google Scholar 

  45. Knippschild U, Milne DM, Campbell LE, DeMaggio AJ, Christenson E, Hoekstra MF, Meek DW (1997) p53 is phosphorylated in vitro and in vivo by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase-directed drugs. Oncogene 15(14):1727–1736

    Article  CAS  PubMed  Google Scholar 

  46. Higashimoto Y, Saito S, Tong XH, Hong A, Sakaguchi K, Appella E, Anderson CW (2000) Human p53 is phosphorylated on serines 6 and 9 in response to DNA damage-inducing agents. J Biol Chem 275(30):23199–23203

    Article  CAS  PubMed  Google Scholar 

  47. Cordenonsi M, Montagner M, Adorno M, Zacchigna L, Martello G, Mamidi A, Soligo S, Dupont S, Piccolo S (2007) Integration of TGF-beta and Ras/MAPK signaling through p53 phosphorylation. Science 315(5813):840–843

    Article  CAS  PubMed  Google Scholar 

  48. Dumaz N, Milne DM, Meek DW (1999) Protein kinase CK1 is a p53-threonine 18 kinase which requires prior phosphorylation of serine 15. FEBS Lett 463(3):312–316

    Article  CAS  PubMed  Google Scholar 

  49. Sakaguchi K, Saito S, Higashimoto Y, Roy S, Anderson CW, Appella E (2000) Damage-mediated phosphorylation of human p53 threonine 18 through a cascade mediated by a casein 1-like kinase. Effect on Mdm2 binding. J Biol Chem 275(13):9278–9283

    Article  CAS  PubMed  Google Scholar 

  50. Shieh SY, Ikeda M, Taya Y, Prives C (1997) DNA damage induced phosphorylation of p53 alleviates inhibition by MDM2. Cell 91:325–334

    Article  CAS  PubMed  Google Scholar 

  51. Saito S, Goodarzi AA, Higashimoto Y, Noda Y, Lees-Miller SP, Appella E, Anderson CW (2002) ATM mediates phosphorylation at multiple p53 sites, including Ser(46), in response to ionizing radiation. J Biol Chem 277:12491–12494

    Article  CAS  PubMed  Google Scholar 

  52. Tibbetts RS, Brumbaugh KM, Williams JM, Sarkaria JN, Cliby WA, Shieh SY, Taya Y, Prives C, Abraham RT (1999) A role for ATR in the DNA damage-induced phosphorylation of p53. Genes Dev 13:152–157

    Article  CAS  PubMed  Google Scholar 

  53. She QB, Chen N, Dong Z (2000) ERKs and p38 kinase phosphorylate p53 protein at serine 15 in response to UV radiation. J Biol Chem 275:20444–20449

    Article  CAS  PubMed  Google Scholar 

  54. MacLaine NJ, Oster B, Bundgaard B, Fraser JA, Buckner C, Lazo PA, Meek DW, Höllsberg P, Hupp TR (2008) A central role for CK1 in catalyzing phosphorylation of the p53 transactivation domain at serine 20 after HHV-6B viral infection. J Biol Chem 283(42):28563–28573

    Article  CAS  PubMed  Google Scholar 

  55. Chehab NH, Malikzay A, Stavridi ES, Halazonetis TD (1999) Phosphorylation of Ser-20 mediates stabilization of human p53 in response to DNA damage. Proc Natl Acad Sci USA 96:13777–13782

    Article  CAS  PubMed  Google Scholar 

  56. Unger T, Juven-Gershon T, Moallem E, Berger M, Vogt Sionov R, Lozano G, Oren M, Haupt Y (1999) Critical role for Ser20 of human p53 in the negative regulation of p53 by Mdm2. EMBO J 18:1805–1814

    Article  CAS  PubMed  Google Scholar 

  57. She QB, Ma WY, Dong Z (2002) Role of MAP kinases in UVB-induced phosphorylation of p53 at serine 20. Oncogene 21:1580–1589

    Article  CAS  PubMed  Google Scholar 

  58. Fields GB, Noble RL (1990) Solid phase peptide synthesis utilizing 9- fluorenylmethoxycarbonyl amino acids. Int J Pept Protein Res 35:161–214

    Article  CAS  PubMed  Google Scholar 

  59. Burzio V, Antonelli M, Allende CC, Allende JE (2002) Biochemical and cellular characteristics of the four splice variants of protein kinase CK1α from zebrafish (Danio rerio). J Cell Biochem 86:805–814

    Article  CAS  PubMed  Google Scholar 

  60. Marin O, Meggio F, Pinna LA (1994) Design and synthesis of two new peptide substrates for the specific and sensitive monitoring of casein kinases-1 and -2. Biochem Biophys Res Commun 198:898–905

    Article  CAS  PubMed  Google Scholar 

  61. Perich JW, Meggio F, Reynolds EC, Marin O, Pinna LA (1992) Role of phosphorylated aminoacyl residues in generating atypical consensus sequences which are recognized by casein kinase-2 but not by casein kinase-1. Biochemistry 31:5893–5897

    Article  CAS  PubMed  Google Scholar 

  62. Molecular operating environment (MOE 2008.10), C.C.G., Inc., 1255 University St., Suite 1600, Montreal, Quebec, Canada H3B 3X3

  63. Cozza G, Moro S, Gotte G (2008) Elucidation of the ribonuclease A aggregation process mediated by 3D domain swapping: a computational approach reveals possible new multimeric structures. Biopolymers 89:26–39

    Article  CAS  PubMed  Google Scholar 

  64. Poletto G, Vilardell J, Marin O, Pagano MA, Cozza G, Sarno S, Falqués A, Itarte E, Pinna LA, Meggio F (2008) The regulatory beta subunit of protein kinase CK2 contributes to the recognition of the substrate consensus sequence. A study with an eIF2 beta-derived peptide. Biochemistry 47:8317–8325

    Article  CAS  PubMed  Google Scholar 

  65. Oleinik NV, Krupenko NI, Krupenko SA (2007) Cooperation between JNK1 and JNK2 in activation of p53 apoptotic pathway. Oncogene 26:7222–7230

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Lorenzo Alberto Pinna.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s00018-010-0622-1

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Venerando, A., Marin, O., Cozza, G. et al. Isoform specific phosphorylation of p53 by protein kinase CK1. Cell. Mol. Life Sci. 67, 1105–1118 (2010). https://doi.org/10.1007/s00018-009-0236-7

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