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Cross talk between protein kinase CK2 and eukaryotic translation initiation factor eIF2β subunit

The carboxyl-terminal domain of eIF2β binds to specific sequences of CK2α which affect its activity

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Abstract

The β-subunit of eukaryotic translation initiation factor eIF2 is a substrate and a partner for protein kinase CK2. Surface plasmon resonance analysis shows that the truncated form corresponding to residues 138–333 of eIF2β (eIF2β-CT) interacts with CK2α as efficiently as full length eIF2β, whereas the form corresponding to residues 1–137, which contains the CK2 phosphorylation sites, (eIF2β-NT) does not bind. The use of different mutants and truncated forms of CK2α allowed us to map the basic segment K74–K83 at the beginning of helix αC and residues R191R195K198 in the p+1 loop as the main determinants for the binding to eIF2β-CT of either the isolated CK2α subunit or the CK2 holoenzyme. The presence of eIF2β-CT stimulated the activity of CK2α towards the RRRAADSDDDDD peptide substrate; effect that was not observed with the CK2α K74-77A whose ability to bind to eIF2β-CT is severely impaired. Gel filtration analysis confirmed the ability of CK2α to form complexes with eIF2β-CT, and the contribution of the basic cluster in CK2α (K74–K77) in this association.

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References

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

    PubMed  Google Scholar 

  2. 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 

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

    PubMed  Google Scholar 

  4. Guerra B, Issinger OG: Protein kinase CK2 and its role in cellular proliferation, development and pathology. Electrophoresis 20: 391–408, 1999

    Google Scholar 

  5. Issinger OG: Phosphorylation of acidic ribosomal proteins from rabbit reticulocytes by a ribosome-associated casein kinase. Biochim Biophys Acta 477: 185–189, 1977

    PubMed  Google Scholar 

  6. Tuazon PT, Merrick WC, Traugh JA: Site-specific phosphorylation of initiation factor 2 by three cyclic nucleotide-independent protein kinases. J Biol Chem 255: 10954–10958, 1980

    PubMed  Google Scholar 

  7. Kimball SR: Eukaryotic initiation factor eIF2. Int. J. Biochem Cell Biol. 31: 25–29, 1999

    PubMed  Google Scholar 

  8. Sonenberg N, Dever TE: Eukaryotic translation initiation factors and regulators. Curr Opinion Struct Biol 13: 56–63, 2003

    Google Scholar 

  9. Laurino JP, Thompson GM, Pacheco E, Castilho BA: The β subunit of eukaryotic initiation factor 2 binds mRNA through the lysine repeats and a region comprising the C2-C2 motif. Mol Cell Biol 19: 173–181, 1999

    PubMed  Google Scholar 

  10. Thompson GM, Pacheco E, Melo EO, Castilho BA: Conserved sequences in the β subunit of archeal and eukaryal translation initiation factor 2 (eIF2), absent from eIF5, mediate interaction with eIF2γ. Biochem J 347: 703–709, 2000

    PubMed  Google Scholar 

  11. Nika J, Rippel S, Hannig EM: Biochemical analysis of the eIF2βγ complex reveals a structural function for eIF2α in catalyzed nucleotide exchange. J Biol Chem 276: 1051–1056, 2001

    PubMed  Google Scholar 

  12. Kimball SR, Heinzinger NK, Horetsky RL, Jefferson LS: Identification of interprotein interactions between the subunits of eukaryotic initiation factors eIF2 and eIF2B. J Biol Chem 273: 3039–3044, 1998

    PubMed  Google Scholar 

  13. Das S, Maiti T, Das K, Maitra U: Specific interaction of eukaryotic translation initiation factor 5 (eIF5) with the β subunit of eIF2. J Biol Chem 272: 31712–31718, 1997

    PubMed  Google Scholar 

  14. Wang X, Paulin FEM, Campbell LE, Gomez E, O’Brien K, Morrice N, Proud CG: Eukaryotic initiation factor 2B: identification of multiple phosphorylation sites in the ɛ-subunit and their functions ‘in vivo’. EMBO J 20: 4349–4359, 2001

    PubMed  Google Scholar 

  15. Majumdar R, Bandyopadhyay A, Deng H, Maitra U: Phosphorylation of mammalian translation initiation factor 5 (eIF5) ‘in vitro’ and ‘in vivo’. Nucleic Acids Res 30: 1154–1162, 2002

    PubMed  Google Scholar 

  16. Llorens F, Roher N, Miro FA, Sarno S, Ruiz FX, Meggio F, Plana M, Pinna LA, Itarte E: Eukaryotic translation-initiation factor eIF2beta binds to protein kinase CK2: effects on CK2alpha activity. Biochem J 375: 623–631, 2003

    PubMed  Google Scholar 

  17. Sarno S, Vaglio P, Marin O, Issinger OG, 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 

  18. Gil C, Plana M, Riera M, Itarte E: Rat liver pp49, a protein that forms complexes with protein kinase CK2, is composed of the β and the γ subunits of translation initiation factor eIF-2. Biochem Biophys Res Commun 225: 1052–1057, 1996

    PubMed  Google Scholar 

  19. Niefind K, Guerra B, Pinna LA, Issinger OG, Schomburg D: Crystal structure of the catalytic subunit of protein kinase CK2 from Zea mays at 2.1 A resolution. EMBO J 17: 2451–2462, 1998

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  22. Miyata Y, Yahara I: Interaction between casein kinase II and the 90-kDa stress protein, HSP90. Biochemistry 34: 8123–8129, 1995

    PubMed  Google Scholar 

  23. Li D, Dobrowolska G, Krebs EG: The physical association of casein kinase 2 with nucleolin. J Biol Chem 271: 15662–15668, 1996

    PubMed  Google Scholar 

  24. Yamaguchi Y, Wada T, Suzuki F, Takagi T, Hasegawa J, Handa H: Casein kinase II interacts with the bZIP domains of several transcription factors. Nucleic Acids Res 26, 3854–3861, 1998

    PubMed  Google Scholar 

  25. Roher N, Sarno S, Miró F, Ruzzene M, Llorens F, Meggio F, Itarte E, Pinna LA, Plana M: The carboxy-terminal domain of Grp94 binds to protein kinase CK2α but not to holoenzyme. FEBS Lett 505: 42–46, 2001

    PubMed  Google Scholar 

  26. Ruzzene M, Brunati AM, Sarno S, Donella-Deana A, Pinna LA: Hematopoietic lineage cell specific protein 1 associates with and down-regulates protein kinase CK2. FEBS Lett 461: 32–36, 1999

    PubMed  Google Scholar 

  27. Ruzzene M, Brunati AM, Sarno S, Marin O, Donella-Deana A, Pinna LA: Ser/Thr phosphorylation of hematopoietic specific protein 1 (HS1). Eur J Biochem 267: 3065–3072, 2000

    PubMed  Google Scholar 

  28. Meggio F, Negro A, Sarno S, Ruzzene M, Bertoli A, Sorgato MA, Pinna L: Bovine prion protein as a modulator of protein kinase CK2. Biochem J 352: 191–196, 2000

    PubMed  Google Scholar 

  29. Asano K, Krishnamoorthy T, Phan L, Pavitt GD, Hinnebusch AG: Conserved bipartite motifs in yeast eIF5 and eIF2Bepsilon, GTPase-activating and GDP-GTP exchange factors in translation initiation, mediate binding to their common substrate eIF2. EMBO J 18: 1673–1688, 1999

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Departament de Bioquímica i Biologia Molecular, Unitat de Bioquímica de Ciències, Universitat Autònoma de Barcelona, Edifici Cs, Campus de Bellaterra, 08193, Bellaterra (Barcelona), Spain

    Franc Llorens, Eduard Sarró, Anna Duarri, Nerea Roher, Maria Plana & Emilio Itarte

  2. Dipartimento di Chimica Biologica, Università di Padova, Padova, Italy

    Stefania Sarno, Flavio Meggio & Lorenzo A. Pinna

  3. Departament de Bioquímica i de Biología Molecular, Unitat de Bioquímica de Ciències, Universitat Autònoma de Barcelona, Edifici Cs, Campus de Bellaterra, 08193, Bellaterra (Barcelona), Spain

    Emilio Itarte

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  1. Franc Llorens
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  2. Stefania Sarno
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  3. Eduard Sarró
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  4. Anna Duarri
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  5. Nerea Roher
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  6. Flavio Meggio
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  7. Maria Plana
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  8. Lorenzo A. Pinna
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  9. Emilio Itarte
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Correspondence to Emilio Itarte.

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Llorens, F., Sarno, S., Sarró, E. et al. Cross talk between protein kinase CK2 and eukaryotic translation initiation factor eIF2β subunit. Mol Cell Biochem 274, 53–61 (2005). https://doi.org/10.1007/s11010-005-3081-5

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  • DOI: https://doi.org/10.1007/s11010-005-3081-5

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