The physical association of casein kinase 2 with nucleolin.
CK2 (formerly called casein kinase 2) is a ubiquitous messenger-independent serine/threonine protein kinase implicated in cell growth and proliferation. To investigate the regulation and functions of this enzyme, experiments were carried out to search for CK2-interacting proteins. The methods employed included an overlay technique, co-purification, co-immunoprecipitation, and the use of glutathione S-transferase (GST) CK2 fusion proteins. By the CK2 overlay technique, one protein of 110 kDa was found to bind to CK2 with very high affinity. The binding was inhibited by CK2 effectors such as heparin, polyarginine, and histone H1, but was not affected by the CK2 substrate, casein. Protein p110 was also detected by co-immunoprecipitation using anti-CK2 antiserum, suggesting an in vivo association of this protein with CK2. Co-purification of p110 with CK2 from Sf-9 cells that overexpressed CK2 was also observed through sequential chromatographic steps. Using GST fusion proteins of CK2, the CK2-p110 interaction was investigated further and was found to occur primarily through CK2 alpha or alpha' subunits, but not the beta subunit. Protein p110 was purified from 3T3 L1 mouse fibroblast cell lines using a GST-CK2 affinity resin. Amino acid sequence analysis of peptides obtained from the protein indicated that it is the nuclear protein, nucleolin. Furthermore, p110 was recognized by anti-nucleolin antiserum. At present, the physiological significance of the strong interaction between CK2 and nucleolin, an excellent substrate for the enzyme, is not clear. However, this association may be important for regulating rDNA transcription.
Li
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Department of Pharmacology, University of Washington, Seattle, Washington 98195, USA.
Dobrowolska
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Krebs
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EG
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D047390:Casein Kinase II; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D016601:RNA-Binding Proteins; D013347:Subcellular Fractions / Q000737:chemistry; D000097796:Nucleolin
D047390:Casein Kinase II; D002364:Caseins; D017931:DNA Primers; D009687:Nuclear Proteins; D010750:Phosphoproteins; D017346:Protein Serine-Threonine Kinases; D016601:RNA-Binding Proteins; D000097796:Nucleolin