Binding of FGF‐1 variants to protein kinase CK2 correlates with mitogenicity

Fibroblast growth factor‐1 (FGF‐1) has both extra‐ and intracellular functions. To identify intracellular binding partners for FGF‐1, we isolated proteins from U2OS human osteosarcoma cells interacting specifically with FGF‐1. One of the isolated proteins was identified as protein kinase CK2 (CK2). We here provide evidence that FGF‐1 binds to both the catalytic α‐subunit and to the regulatory β‐subunit of CK2. The interaction between FGF‐1 and CK2α and β was characterized by surface plasmon resonance, giving KD values of 0.4 ± 0.3 and 1.2 ± 0.2 μM, respectively. By using a novel assay for intracellular protein interaction, FGF‐1 and CK2α are shown to interact in vivo. In vitro, FGF‐1 and FGF‐2 are phosphorylated by CK2, and the presence of FGF‐1 or FGF‐2 was found to enhance the autophosphorylation of CK2β. A correlation between the mitogenic potential of FGF‐1 mutants and their ability to bind to CK2α was observed. The possible involvement of CK2 in the FGF‐induced stimulation of DNA synthesis is discussed.

[1]  D. Moscatelli,et al.  The FGF family of growth factors and oncogenes. , 1992, Advances in cancer research.

[2]  F. Amalric,et al.  Uncoupling of cell proliferation and differentiation activities of basic fibroblast growth factor , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  J. Goodman,et al.  An oligomeric protein is imported into peroxisomes in vivo , 1994, The Journal of cell biology.

[4]  G. Sheu,et al.  Effects of autophosphorylation on casein kinase II activity: evidence from mutations in the beta subunit. , 1994, Biochemistry.

[5]  J. Wesche,et al.  Identification of Ribosome-binding Protein p34 as an Intracellular Protein That Binds Acidic Fibroblast Growth Factor* , 2002, The Journal of Biological Chemistry.

[6]  J. Teissié,et al.  Basic fibroblast growth factor enters the nucleolus and stimulates the transcription of ribosomal genes in ABAE cells undergoing G0----G1 transition. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Feige,et al.  Basic fibroblast growth factor is a substrate for protein phosphorylation and is phosphorylated by capillary endothelial cells in culture. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Jaye,et al.  Possible dissociation of the heparin-binding and mitogenic activities of heparin-binding (acidic fibroblast) growth factor-1 from its receptor-binding activities by site-directed mutagenesis of a single lysine residue , 1990, The Journal of cell biology.

[9]  S. Yu,et al.  Protein kinase CK2 signal in neoplasia. , 2001, Histology and histopathology.

[10]  Jordan T. Shin,et al.  The HIV-1 TAT Protein Induces the Expression and Extracellular Appearance of Acidic Fibroblast Growth Factor (*) , 1995, The Journal of Biological Chemistry.

[11]  M. Montenarh,et al.  Subcellular localization of protein kinase CK2 , 2000, Cell and Tissue Research.

[12]  F. Amalric,et al.  Fibroblast Growth Factor-2 Binds to the Regulatory β Subunit of CK2 and Directly Stimulates CK2 Activity toward Nucleolin* , 1996, The Journal of Biological Chemistry.

[13]  M. Montenarh,et al.  Binding domain for p21(WAF1) on the polypeptide chain of the protein kinase CK2 beta-subunit. , 2000, Biochemical and biophysical research communications.

[14]  T. Maciag,et al.  The heparin-binding (fibroblast) growth factor family of proteins. , 1989, Annual review of biochemistry.

[15]  M. Montenarh,et al.  Subcellular localization of protein kinase CK2. A key to its function? , 2000, Cell and tissue research.

[16]  D. Litchfield,et al.  Expression and localization of epitope‐tagged protein kinase CK2 , 1997, Journal of cellular biochemistry.

[17]  T. Hla,et al.  Recovery of mitogenic activity of a growth factor mutant with a nuclear translocation sequence. , 1990, Science.

[18]  J. Fallon,et al.  Fibroblast growth factors as multifunctional signaling factors. , 1999, International review of cytology.

[19]  C. Allende,et al.  Promiscuous subunit interactions: a possible mechanism for the regulation of protein kinase CK2. , 1998, Journal of cellular biochemistry. Supplement.

[20]  J A Thompson,et al.  Serum-starvation induces the extracellular appearance of FGF-1. , 1996, Biochimica et biophysica acta.

[21]  J. Winkles,et al.  Structure‐function studies of heparin‐binding (acidic fibroblast) growth factor‐1 using site‐directed mutagenesis , 1991, Journal of cellular biochemistry.

[22]  International Human Genome Sequencing Consortium.  Initial sequencing and analysis of the human genome , 2001, Nature.

[23]  Thomas Maciag,et al.  The Extravesicular Domain of Synaptotagmin-1 Is Released with the Latent Fibroblast Growth Factor-1 Homodimer in Response to Heat Shock* , 1998, The Journal of Biological Chemistry.

[24]  C. Allende,et al.  Protein kinase CK2: an enzyme with multiple substrates and a puzzling regulation , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  E. Chambaz,et al.  A role for protein kinase CK2 in cell proliferation: evidence using a kinase-inactive mutant of CK2 catalytic subunit α , 2001, Oncogene.

[26]  Honey Chan,et al.  Acyl-CoA oxidase is imported as a heteropentameric, cofactor-containing complex into peroxisomes of Yarrowia lipolytica , 2002, The Journal of cell biology.

[27]  J. Hériché,et al.  Regulation of protein phosphatase 2A by direct interaction with casein kinase 2alpha. , 1997, Science.

[28]  Kirsten Sandvig,et al.  Dual mode of signal transduction by externally added acidic fibroblast growth factor , 1994, Cell.

[29]  S. Mische,et al.  Routine identification of proteins from sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) gels or polyvinyl difluoride membranes using matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) , 1998, Electrophoresis.

[30]  L Andrews,et al.  An improved procedure for enzymatic digestion of polyvinylidene difluoride-bound proteins for internal sequence analysis. , 1994, Analytical biochemistry.

[31]  S. Olsnes,et al.  Effects of Mutations of a Phosphorylation Site in an Exposed Loop in Acidic Fibroblast Growth Factor* , 1999, The Journal of Biological Chemistry.

[32]  I. Mason.  The ins and outs of fibroblast growth factors , 1994, Cell.

[33]  H. Stenmark,et al.  Cloning of an intracellular protein that binds selectively to mitogenic acidic fibroblast growth factor. , 1998, The Biochemical journal.

[34]  J. Schlessinger,et al.  Measurement of the binding of tyrosyl phosphopeptides to SH2 domains: a reappraisal. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[35]  A. Rapak,et al.  Stimulation of proliferation of a human osteosarcoma cell line by exogenous acidic fibroblast growth factor requires both activation of receptor tyrosine kinase and growth factor internalization , 1996, Molecular and cellular biology.

[36]  F. Pancetti,et al.  Heterogeneous nuclear ribonucleoprotein A2 interacts with protein kinase CK2. , 1999, Biochemical and biophysical research communications.

[37]  L. Pinna,et al.  Protein kinase CK2 ("casein kinase-2") and its implication in cell division and proliferation. , 1997, Progress in cell cycle research.

[38]  M. Jaye,et al.  Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation , 1994, Cell.

[39]  O. Issinger,et al.  CK2: a protein kinase in need of control. , 1999, Pharmacology & therapeutics.

[40]  S. Subramani,et al.  Import of stably folded proteins into peroxisomes. , 1995, Molecular biology of the cell.

[41]  M. Czech,et al.  Enhanced casein kinase II activity in COS-1 cells upon overexpression of either its catalytic or noncatalytic subunit. , 1991, The Journal of biological chemistry.

[42]  N. Quarto,et al.  Fibroblast growth factor-2 (FGF-2) in the nucleus: translocation process and targets. , 1994, Biochemical pharmacology.

[43]  A. Rapak,et al.  Inability of the Acidic Fibroblast Growth Factor Mutant K132E to Stimulate DNA Synthesis after Translocation into Cells* , 1998, The Journal of Biological Chemistry.

[44]  L. Pinna,et al.  Bovine prion protein as a modulator of protein kinase CK2. , 2000, The Biochemical journal.

[45]  F. Tarantini,et al.  S100A13 Is Involved in the Regulation of Fibroblast Growth Factor-1 and p40 Synaptotagmin-1 Release in Vitro * , 1998, The Journal of Biological Chemistry.

[46]  R. Forough,et al.  Heat shock induces the release of fibroblast growth factor 1 from NIH 3T3 cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[47]  W. Pyerin.  Human casein kinase II: structures, genes, expression and requirement in cell growth stimulation. , 1994, Advances in enzyme regulation.

[48]  A. Rapak,et al.  Translocation to Cytosol of Exogenous, CAAX-tagged Acidic Fibroblast Growth Factor (*) , 1995, The Journal of Biological Chemistry.