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Yeast holoenzyme of protein kinase CK2 requires both β and β′ regulatory subunits for its activity

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Abstract

Protein kinase CK2 is a highly conserved Ser/Thr protein kinase that is ubiquitous among eucaryotic organisms and appears to play an important role in many cellular functions. This enzyme in yeast has a tetrameric structure composed of two catalytic (α and/or α′) subunits and two regulatory β and β′ subunits. Previously, we have reported isolation from yeast cells four active forms of CK2, composed of αα′ββ′, α2ββ′, α′2ββ′ and a free α′-catalytic subunit. Now, we report that in Saccharomyces cerevisiae CK2 holoenzyme regulatory β subunit cannot substitute other β′ subunit and only both of them can form fully active enzymatic unit. We have examined the subunit composition of tetrameric complexes of yeast CK2 by transformation of yeast strains containing single deletion of the β or β′ regulatory subunits with vectors carrying lacking CKB1 or CKB2 genes. CK2 holoenzyme activity was restored only in cases when both of them were present in the cell. Additional, co-immunoprecypitation experiments show that polyadenylation factor Fip1 interacts with catalytic α subunits of CK2 and interaction with beta subunits in the holoenzyme decreases CK2 activity towards this protein substrate. These data may help to elucidate the role of yeast protein kinase CK2β/β′ subunits in the regulation of holoenzyme assembly and phosphotransferase activity.

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Abbreviations

hCK2:

holoenzyme of protein kinase CK2 (casein kinase type 2)

rCK2α and rCK2α′:

recombinants of CK2α and CK2α′ respectively

rP2B:

recombinant of yeast 60S protein P2B

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Acknowledgments

This work was supported by scientific resources for years 2005–2008 as a ordered research project PBZ-MIN-014/P05/2004 from Ministry of Education and Science, and partially by grant from Catholic University of Lublin.

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Authors and Affiliations

  1. Department of Molecular Biology, Environmental Protection Institute, John Paul II Catholic University of Lublin, Kraśnicka Av. 102, 20-718, Lublin, Poland

    Konrad Kubiński, Katarzyna Domańska, Ewa Sajnaga, Elżbieta Mazur, Rafał Zieliński & Ryszard Szyszka

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  1. Konrad Kubiński
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  2. Katarzyna Domańska
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  3. Ewa Sajnaga
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  4. Elżbieta Mazur
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  5. Rafał Zieliński
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  6. Ryszard Szyszka
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Correspondence to Ryszard Szyszka.

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Kubiński, K., Domańska, K., Sajnaga, E. et al. Yeast holoenzyme of protein kinase CK2 requires both β and β′ regulatory subunits for its activity. Mol Cell Biochem 295, 229–236 (2007). https://doi.org/10.1007/s11010-006-9292-6

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  • DOI: https://doi.org/10.1007/s11010-006-9292-6

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