Abstract
Protein kinase CK2 has traditionally been described as a stable heterotetrameric complex (α < eqid1 > β2) but new approaches that effectively capture the dynamic behavior of proteins, are bringing a new picture of this complex into focus. To track the spatio-temporal dynamics of CK2 in living cells, we fused its catalytic α and regulatory β subunits with GFP and analog proteins. Beside the mostly nuclear localization of both subunits, and the identification of specific domains on each subunit that triggers their localization, the most significant finding was that the association of both CK2 subunits in a stable tetrameric holoenzyme eliminates their nuclear import (Mol Cell Biol {23}: 975–987, 2003). Molecular movements of both subunits in the cytoplasm and in the nucleus were analyzed using different new and updated fluorescence imaging methods such as: fluorescence recovery after photo bleaching (FRAP), fluorescence loss in photo bleaching (FLIP), fluorescence correlation spectroscopy (FCS), and photoactivation using a biphoton microscope. These fluorescence-imaging techniques provide unprecedented ways to visualize and quantify the mobility of each individual CK2 subunit with high spatial and temporal resolution. Visualization of CK2 heterotetrameric complex formation could also be recorded using the fluorescence resonance energy transfer (FRET) technique. FRET imaging revealed that the assembling of this molecular complex can take place both in the cytoplasmic and nuclear compartments. The spatio–temporal organization of individual CK2 subunits and their dynamic behavior remain now to be correlated with the functioning of this kinase in the complex environment of the cell.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Filhol O, Nueda A, Martel V, Gerber-Scokaert D, Benitez MJ, Souchier C, Saoudi Y, Cochet C: Live-cell fluorescence imaging reveals the dynamics of protein kinase CK2 individual subunits. Mol Cell Biol 23: 975–987, 2003
Ahmed K, Gerber DA, Cochet C: Joining the cell survival squad: An emerging role for protein kinase CK2. Trends Cell Biol 12: 226–230, 2002
Litchfield DW: Protein kinase CK2: Structure, regulation and role in cellular decisions of life and death. Biochem J 369: 1–15, 2003
Guerra B, Siemer S, Boldyreff B, Issinger OG: Protein kinase CK2: evidence for a protein kinase CK2beta subunit fraction, devoid of the catalytic CK2alpha subunit, in mouse brain and testicles. FEBS Lett 462: 353–357, 1999
Filhol O, Martiel JL, Cochet C: Protein kinase CK2: A new view of an old molecular complex. EMBO Rep 5: 351–355, 2004
Lippincott-Schwartz J, Patterson GH: Development and use of fluorescent protein markers in living cells. Science 300: 87–91, 2003
van Roessel P, Brand AH: Imaging into the future: Visualizing gene expression and protein interactions with fluorescent proteins. Nat Cell Biol 4: E15–E20, 2002
Patterson GH, Lippincott-Schwartz J: A photoactivatable GFP for selective photolabeling of proteins and cells. Science 297: 1873–1877, 2002
Patterson GH, Lippincott-Schwartz J: Selective photolabeling of proteins using photoactivatable GFP. Methods 32: 445–450, 2004
Wouters FS, Bastiaens PI, Wirtz KW, Jovin TM, Bastiaens PH, Wanders RJ, Seedorf U: FRET microscopy demonstrates molecular association of non-specific lipid transfer protein (nsL-TP) with fatty acid oxidation enzymes in peroxisomes.The non-specific lipid transfer protein (sterol carrier protein 2) acts as a peroxisomal fatty acyl-CoA binding protein. EMBO J 17: 7179–7189, 1998
Martel V, Filhol O, Nueda A, Gerber D, Benitez MJ, Cochet C: Visualization and molecular analysis of nuclear import of protein kinase CK2 subunits in living cells. Mol Cell Biochem 227: 81–90, 2001
Martel V, Filhol O, Nueda A, Cochet C: Dynamic localization/association of protein kinase CK2 subunits in living cells: a role in its cellular regulation? Ann N Y Acad Sci 973: 272–277, 2002
Chantalat L, Leroy D, Filhol O, Nueda A, Benitez MJ, Chambaz EM, Cochet C, Dideberg O: Crystal structure of the human protein kinase CK2 regulatory subunit reveals its zinc finger-mediated dimerization. EMBO J 18: 2930–2940, 1999
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
Glover CV: A filamentous form of Drosophila casein kinase II. J Biol Chem 261: 14349–14354, 1986
Valero E, De Bonis S, Filhol O, Wade RH, Langowski 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
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Theis-Febvre, N., Martel, V., Laudet, B. et al. Highlighting protein kinase CK2 movement in living cells. Mol Cell Biochem 274, 15–22 (2005). https://doi.org/10.1007/s11010-005-3115-z
Issue Date:
DOI: https://doi.org/10.1007/s11010-005-3115-z