Molecular and Cellular Biochemistry

, Volume 353, Issue 1–2, pp 81–91 | Cite as

The cytosolic protein kinase CK2 phosphorylates cardiac calsequestrin in intact cells

  • Timothy P. McFarland
  • Naama H. Sleiman
  • Daniel B. Yaeger
  • Steven E. Cala


The luminal SR protein CSQ2 contains phosphate on roughly half of the serines found in its C-terminus. The sequence around phosphorylation sites in CSQ2 suggest that the in vivo kinase is protein kinase CK2, even though this enzyme is thought to be present only in the cytoplasm and nucleus. To test whether CSQ2 kinase is CK2, we combined approaches that reduced CK2 activity and CSQ2 phosphorylation in intact cells. Tetrabromocinnamic acid, a specific inhibitor of CK2, inhibited both the CSQ2 kinase and CK2 in parallel across a range of concentrations. In intact primary adult rat cardiomyocytes and COS cells, 24 h of drug treatment reduced phosphorylation of overexpressed CSQ2 by 75%. Down-regulation of CK2α subunits in COS cells using siRNA, produced a 90% decrease in CK2α protein levels, and CK2-silenced COS cells exhibited a twofold reduction in CSQ2 kinase activity. Phosphorylation of CSQ2 overexpressed in CK2-silenced cells was also reduced by a factor of two. These data suggested that CSQ2 in intact cells is phosphorylated by CK2, a cytosolic kinase. When phosphorylation site mutants were analyzed in COS cells, the characteristic rough endoplasmic reticulum form of the CSQ2 glycan (GlcNAc2Man9,8) underwent phosphorylation site dependent processing such that CSQ2-nonPP (Ser to Ala mutant) and CSQ2-mimPP (Ser to Glu mutant) produced apparent lower and greater levels of ER retention, respectively. Taken together, these data suggest CK2 can phosphorylate CSQ2 co-translationally at biosynthetic sites in rough ER, a process that may result in changes in its subsequent trafficking through the secretory pathway.


Calsequestrin CK2 Phosphorylation Sarcoplasmic reticulum Kinase inhibitors siRNA Glycosylation Rough ER 



Cardiac calsequestrin


Fast-twitch skeletal muscle calsequestrin


Wild-type cardiac calsequestrin sequence


Sarcoplasmic reticulum


Endoplasmic reticulum


CSQ-WT adenovirus


Adenovirus for CSQ2 Ser → Ala mutant


adenovirus for CSQ2 Ser → Glu mutant


Multiplicity of infection


Tetrabromocinnamic acid







We wish to thank Lauren Dovantzis for expert technical assistance. This study was supported by grant HL62586 from the NIH/NHLBI.


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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Timothy P. McFarland
    • 1
  • Naama H. Sleiman
    • 1
  • Daniel B. Yaeger
    • 1
  • Steven E. Cala
    • 1
  1. 1.Department of PhysiologyWayne State University School of MedicineDetroitUSA

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