Casein kinase II accumulation in the nucleolus and its role in nucleolar phosphorylation.
A rabbit antiserum against highly purified casein kinase II from mouse tumor cells was used for immunolocalization of the enzyme in fixed, permeabilized mouse cells. Casein kinase II was highly accumulated in nucleoli compared to the extra-nucleolar space of the nucleus or to the cytoplasma. Casein kinase II samples highly purified from the cytoplasma, from the extra-nucleolar fraction of the nucleus or from nucleoli exhibited no differences with respect to structure and function. All samples originally had an alpha 2 beta 2 structure (alpha, 42 kDa; beta, 24 kDa) showing formation of the alpha'-chain (36 kDa) only in the late steps of purification. The isoelectric point of the alpha-chain of all three samples was pH 7.7 and that of the beta-chain was pH 6.4-6.6. Using ATP or GTP, all three casein kinase II samples gave the same results of maximum phosphorylation of purified nucleolar marker phosphoproteins pp105/C23, pp135 and B23, yielding pp135 as one of the most highly phosphorylated proteins with an incorporation of about 75 phosphate groups per molecule pp135. Studies on optimum conditions of phosphorylation of nucleolar phosphoproteins by casein kinase II revealed that each of the protein substrates individually responded to alterations of assay parameters such as pH, magnesium ion and sodium chloride concentrations indicating that predominantly individual structural criteria were responsible for optimum phosphorylation. The determination of the apparent Km of casein kinase II for purified nucleolar phosphoproteins yielded values of 0.15 microM (pp105/C23), 0.1 microM (pp135) and 1.0 microM (B23) identifying them as high-affinity substrates of casein kinase II.
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Friedrich-Miescher-Laboratorium der Max-Planck-Gesellschaft, Tübingen, F.R.G.
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D000818:Animals; D047388:Casein Kinases; D002460:Cell Line; D002466:Cell Nucleolus / Q000201:enzymology* / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D007158:Immunologic Techniques; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011073:Polyamines / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism*; D011794:Quercetin / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology
D009687:Nuclear Proteins; D010750:Phosphoproteins; D011073:Polyamines; D006493:Heparin; D011794:Quercetin; D011494:Protein Kinases; D047388:Casein Kinases