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Autophosphorylation at the regulatory β subunit reflects the supramolecular organization of protein kinase CK2

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Abstract

Among the features of protein kinase CK2, autophosphorylation at its β-subunit(s) upon incubation with ATP/Mg++ was early detected as a rapid and stoichiometric event occurring through an intramolecular mechanism as judged from kinetic analyses. The autophosphorylation site was mapped to Ser2 and, to a lesser extent, Ser3 both fulfilling the CK2 consensus sequence (MSSSEEV). The crystal structure of the heterotetrameric holoenzyme, however, is not compatible with an intramolecular autophosphorylation of the N-terminal stretch of either of the two β subunits. Here we show that efficient “intramolecular” autophosphorylation of the β subunit is crucially dependent on the formation of oligomers composed by several holoenzyme heterotetrameric protomers. Increasing ionic strength of the incubation medium promoting dissociation of the supramolecular oligomers abrogates β subunit autophosphorylation, although CK2 catalytic activity, as judged from the phosphorylation of exogenous substrates, is still quite evident. These findings, in conjunction with graphic modelization, support the view that CK2 autophosphorylation at its β subunits takes place through an “intraoligomeric” mechanism where the β subunits of a protomer are phosphorylated by the catalytic subunits of another adjacent protomer. It appears therefore that in vivo β autophosphorylation is symptomatic of supramolecular CK2 oligomers.

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Abbreviations

CK2:

casein kinase 2

nCK2:

native CK2

rCK2:

recombinant human CK2

References

  1. Pinna LA: Casein kinase 2: An “eminence grise” in cellular regulation? Biochim Biophys Acta 1054: 267–284, 1990

    PubMed  Google Scholar 

  2. Pinna LA: Protein kinase CK2: a challenge to canons. J Cell Sci 115: 3873–3878, 2002

    PubMed  Google Scholar 

  3. Pinna LA: The raison d’etre of constitutively active protein kinase CK2. Acc Chem Res 36: 378–384, 2003

    PubMed  Google Scholar 

  4. Allende JE, Allende CC: Protein kinase CK2: an enzyme with multiple substrates and a puzzling regulation. FASEB J 9: 313–323, 1995

    PubMed  Google Scholar 

  5. Guerra B, Issinger O-G: Protein kinase CK2 and its role in cellular proliferation, development and pathology. Electrophoresis 20: 391–408, 1999

    PubMed  Google Scholar 

  6. Litchfield DW: Protein kinase CK2: structure, regulation and role in cellular decisions of life and death. Biochem J 369: 1–15, 2003

    PubMed  Google Scholar 

  7. Meggio F, Pinna LA: One-thousand-and-one substrates of protein kinase CK2? FASEB J 17: 349–368, 2003

    PubMed  Google Scholar 

  8. Stigare J, Buddelmeijer N, Pigon A, Egyhazi E: A majority of casein kinase IIα subunit is tightly bound to intranuclear components but not to the β subunit. Mol Cell Biochem 129: 77–85, 1993

    PubMed  Google Scholar 

  9. Guerra B, Siemer S, Boldyreff B, Issinger O-G: Protein kinase CK2: evidence for a protein kinase CK2β subunit fraction, devoid of the catalytic CK2α subunit, in mouse brain and testicles. FEBS Lett 462: 353–357, 1999

    PubMed  Google Scholar 

  10. Faust M, Schuster N, Montenarh M: Specific binding of protein kinase CK2 catalytic subunits to tubulin. FEBS Lett 462: 51–56, 1999

    PubMed  Google Scholar 

  11. Meggio F, Boldyreff B., Marin O, Pinna LA, Issinger O-G: Role of the β subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme. Eur J Biochem 204: 293–297, 1992

    PubMed  Google Scholar 

  12. Boldyreff B, Meggio F, Pinna LA, Issinger O-G: Reconstitution of normal and hyperactivated forms of casein kinase-2 by variably mutated β-subunits. Biochemistry 32: 12672–12677, 1993

    PubMed  Google Scholar 

  13. Antonelli M, Daniotti JL, Rojo D, Allende CC, Allende JE: Cloning, expression and properties of the α′ subunit of casein kinase 2 from zebrafish (Danio rerio). Eur J Biochem 241: 272–279, 1996

    PubMed  Google Scholar 

  14. Marin O, Sarno S, Boschetti M, Pagano MA, Meggio F, Ciminale V, D’Agostino DM, Pinna LA: Unique features of HIV-1 Rev protein phosphorylation by protein kinase CK2 (casein kinase-2). FEBS Lett 481: 63–67, 1999

    Google Scholar 

  15. Grein S, Reymond K, Cochet C, Pyerin W, Chambaz EM, Filhol O: Searching interaction partners of protein kinase CK2β subunit by two-hybrid screening. Mol Cell Biochem 191: 105–109, 1999

    PubMed  Google Scholar 

  16. Graham KC, Litchfield DW: The regulatory β subunit of protein kinase CK2 mediates formation of tetrameric CK2 complexes. J Biol Chem 275: 5003–5010, 2000

    PubMed  Google Scholar 

  17. Boldyreff B, James P, Staudenmann W, Issinger O-G: Ser2 is the autophosphorylation site in the β subunit from bicistronically expressed human casein kinase-2 and from rat liver casein kinase-2β. Eur J Biochem 218: 515–521, 1993

    PubMed  Google Scholar 

  18. Litchfield DW, Lozeman FJ, Cicirelli MF, Harrylock M, Ericsson LH, Piening CJ, Krebs EG: Phosphorylation of the β subunit of casein kinase II in human A431 cells. Identification of the autophosphorylation site and a site phosphorylated by p34cdc2. J Biol Chem 266: 20380–20389, 1991

    PubMed  Google Scholar 

  19. Lin W-JL, Sheu G-T, Traugh JA: Effects of autophosphorylation on casein kinase II activity: evidence from mutations in the β subunit. Biochemistry 33: 6998–7004, 1994

    PubMed  Google Scholar 

  20. Boldyreff B, Meggio F, Pinna LA, Issinger O-G: Efficient autophosphorylation and phosphorylation of the β-subunit by casein kinase-2 require the integrity of an acidic cluster 50 residues downstream from the phosphoacceptor site. J Biol Chem 269: 4827–4831, 1994

    PubMed  Google Scholar 

  21. Bodenbach L, Fauss J, Robitzki A, Krehan A, Lorenz P, Lozeman FJ, Pyerin W: Recombinant human casein kinase II. A study with the complete set of subunits (α, α′ and β), site-directed autophosphorylation mutants and a bicistronically expressed holoenzyme. Eur J Biochem 220: 263–273, 1994

    PubMed  Google Scholar 

  22. Meggio F, Pinna LA: Subunit structure and autophosphorylation mechanism of casein kinase-TS (type-2) from rat liver cytosol. Eur J Biochem 145: 593–599, 1984

    PubMed  Google Scholar 

  23. Niefind K, Guerra B, Ermakowa I, Issinger O-G: Crystal structure of human protein kinase CK2: insights into basic properties of the CK2 holoenzyme. EMBO J 20: 5320–5331, 2001

    PubMed  Google Scholar 

  24. Meggio F, Donella Deana A, Pinna LA: Endogenous phosphate acceptor proteins for rat liver cytosolic casein kinases. J Biol Chem 256: 11958–11961, 1981

    PubMed  Google Scholar 

  25. Sarno S, Vaglio P, Meggio F, Issinger O-G, Pinna LA: Protein kinase CK2 mutants defective in substrate recognition. Purification and kinetic analysis. J Biol Chem 271: 10595–10601, 1996

    PubMed  Google Scholar 

  26. Ruzzene M, Pinna LA: Assay of protein kinases and phosphatases using specific peptide substrates. In: D.G. Hardie (ed.). Protein Phosphorylation – A practical Approach. Oxford University Press, London, 1999, pp 221–253

    Google Scholar 

  27. Ausiello G, Cesareni G, Helmer Citterich M: ESCHER: a new docking procedure applied to the reconstruction of protein tertiary structure. Proteins 28: 556–567, 1997

    PubMed  Google Scholar 

  28. Rekha N, Srinivasan N: Structural basis and substrate specificity of protein kinase CK2 deduced from the modeling of protein-protein interactions. BMC Struct Biol 3(1): 4–17, 2003

    PubMed  Google Scholar 

  29. Sarno S, Vaglio P, Marin O, Issinger O-G, Ruffato K, Pinna LA: Mutational analysis of residues implicated in the interaction between protein kinase CK2 and peptide substrates. Biochemistry 36: 11717–11724, 1997

    PubMed  Google Scholar 

  30. {Zhang C, Vilk G, Canton DA, Litchfield DW: Phosphorylation regulates the stability of the regulatory CK2β subunit. Oncogene 21: 3754–3764, 2002

    PubMed  Google Scholar 

  31. Glover, CG: A filamentous form of Drosophila casein kinase II. J Biol Chem 261: 14349–14354, 1986

    Google Scholar 

  32. Valero E, De Bonis S, Filhol O, Wade RH, Lanhowski J, Chambaz EM, Cochet C: Quaternary structure of casein kinase 2. Characterization of multiple oligomeric states and relation with its catalytic activity. J Biol Chem 270: 8345–8352, 1995

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Dipartimento di Chimica Biologica, Università di Padova, viale G. Colombo 3, 35121, Padova, Italy

    Mario A. Pagano, Stefania Sarno, Giorgia Poletto, Giorgio Cozza, Lorenzo A. Pinna & Flavio Meggio

  2. Dipartimento di Chimica Biologica, Viale G. Colombo, 3, 35121, Padova, Italy

    Flavio Meggio

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  1. Mario A. Pagano
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  2. Stefania Sarno
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  3. Giorgia Poletto
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  4. Giorgio Cozza
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  5. Lorenzo A. Pinna
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  6. Flavio Meggio
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Correspondence to Flavio Meggio.

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Pagano, M.A., Sarno, S., Poletto, G. et al. Autophosphorylation at the regulatory β subunit reflects the supramolecular organization of protein kinase CK2. Mol Cell Biochem 274, 23–29 (2005). https://doi.org/10.1007/s11010-005-3116-y

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