Molecular and Cellular Biochemistry

, Volume 312, Issue 1–2, pp 61–69

Yeast surviving factor Svf1 as a new interacting partner, regulator and in vitro substrate of protein kinase CK2

  • Maciej Masłyk
  • Elżbieta Kochanowicz
  • Rafał Zieliński
  • Konrad Kubiński
  • Ulf Hellman
  • Ryszard Szyszka
Article

DOI: 10.1007/s11010-008-9721-9

Cite this article as:
Masłyk, M., Kochanowicz, E., Zieliński, R. et al. Mol Cell Biochem (2008) 312: 61. doi:10.1007/s11010-008-9721-9

Abstract

Since Svf1 is phosphoprotein, we investigated whether it was a substrate for protein kinase CK2. According to the amino acid sequence Svf1 harbours 20 putative CK2 phosphorylation sites. Here, we have reported cloning, overexpression, purification and characterization of yeast Svf1 as a substrate for three forms of yeast CK2. Svf1 serves as a substrate for both the recombinant CK2α (Km 0.35 μM) and CK2α′ (Km 0.18 μM) as well as CK2 holoenzyme (Km 1.1 μM). Different Km values argue that CK2β(β′) subunit has an inhibitory effect on the activity of both CK2α and CK2α′ towards surviving factor Svf1. Reconstitution of α′2ββ′ isoform of CK2 holoenzyme shows that β/β′ subunits have regulatory effect depending on the kind of CK2 catalytic subunit. This effect was not observed in the case of α2ββ′ isoform, which may be due to interaction between Svf1 and regulatory CK2β subunit (shown by co-immunoprecipitation experiments). Interactions between CK2 subunits and Svf1 protein may have influence on ATP as well as ATP-competitive inhibitors (TBBt and TBBz) binding. CK2 phosphorylates up to six serine residues in highly acidic peptide K199EVIPESDEEESSADEDDNEDEDEESGDSEEESGSEEESDSEEVEITYED248 of the Svf1 protein in vitro. Presented data may help to elucidate the role of protein kinase CK2 and Svf1 in the regulation of cell survival pathways.

Keywords

Yeast Protein kinase CK2 Phosphorylation Apoptosis Svf1 Mass spectrometry Svf1–CK2β interaction ATP-competitive inhibitors 

Abbreviations

hCK2

Holoenzyme of protein kinase CK2 (casein kinase type 2)

rCK2α, rCK2α′, rCK2β and rCK2β′

Recombinant proteins of catalytic: CK2α and CK2α′, and regulatory subunits CK2β and CK2β′, respectively

TBBt

4,5,6,7-Tetrabromo-1H-benzotriazole

TBBz

4,5,6,7-Tetrabromo-1H-benzimidazole

Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Maciej Masłyk
    • 1
  • Elżbieta Kochanowicz
    • 1
  • Rafał Zieliński
    • 1
  • Konrad Kubiński
    • 1
  • Ulf Hellman
    • 2
  • Ryszard Szyszka
    • 1
  1. 1.Department of Molecular Biology, Environmental Protection InstituteJohn Paul II Catholic University of LublinLublinPoland
  2. 2.Ludwig Institute for Cancer ResearchUppsalaSweden

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