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Isolation of an Arabidopsis thaliana casein kinase II β subunit by complementation in Saccharomyces cerevisiae

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Abstract

Casein kinase II is thought to play an essential role in the control of cell division and differentiation in all eukaryotes. Through complementation of a defective casein kinase II catalytic subunit gene from Saccharomyces cerevisiae, we isolated an Arabidopsis thaliana casein kinase II regulatory subunit homologue, CKB1. A second regulatory subunit was identified by low-stringency hybridization with CKB1.

Casein kinase II from S. cerevisiae is composed of two catalytic (α) and two regulatory (β) subunits. Simultaneous disruption of the genes for the α and α′ subunits, CKA1 and CKA2, respectively, is lethal. Strain YDH8 has disruptions of CKA1 and CKA2; its viability depends on a temperature-sensitive allele of CKA2, cka2–8, carried on a centromeric plasmid. We screened an A. thaliana cDNA library, whose inserts are under the control of the galactose-inducible GAL10 promoter, for cDNAs which enabled YDH8 cells to grow at the restrictive temperature. One cDNA, CKB1, was isolated by this screen which had homology to cDNAs of casein kinase II β subunits. A second cDNA, CKB2, was isolated by hybridization and was also able to suppress the YDH8 mutant phenotype.

The proteins encoded by CKB1 and CKB2 are 80% identical. The carboxy-terminal two thirds of both proteins is ca. 54% identical to the regulatory β subunits of casein kinase II from other species. The amino termini are unrelated to any other known proteins. CKB1 and CKB2 lack the conserved autophosphorylation site characteristic of animal β subunits, but have potential casein kinase II phosphorylation sites in the same region. Suppression of the cka1 Δ cka2–8 mutant phenotype occurs by interaction of CKB1 with the defective, cka2–8-encoded, catalytic subunit. Cells with disruptions in CKA1 and CKA2 are not rescued by expression of CKB1.

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References

  1. Bidwai AP, Hanna DE, Glover CVC: Purification and characterization of casein kinase II (CKII) from Δcka1 Δcka2 Saccharomyces cerevisiae rescued by Drosophila CKII subunits. J Biol Chem 267: 18790–18796 (1992).

    Google Scholar 

  2. Bidwai AP, Reed JC, Glover CVC: Phosphorylation of calmodulin by the catalytic subunit of casein kinase II is inhibited by the regulatory subunit. Arch Biochem Biophys 300: 265–270 (1993).

    Google Scholar 

  3. Bidwai AP, Reed JC, Glover CVC: Subunit composition of casein kinase II from S. cerevisae: evidence that the yeast enzyme contains two distinct regulatory beta subunits. J Cell Biochem (S17A): 274–274 (1993).

    Google Scholar 

  4. Boldyreff B, Meggio F, Dobrowolska G, Pinna LA, Issinger O-G: Expression and characterization of a recombinant maize CK-2 α subunit. Biochim Biophys Acta 1173: 32–38 (1993).

    Google Scholar 

  5. Boldyreff B, Meggio F, Pinna LA, Issinger O-G: Casein kinase-2 structure-function relationship: creation of a set of mutants of the β subunit that variably surrogate the wild type β subunit function. Biochem Biophys Res Comm 188: 228–234 (1992).

    Google Scholar 

  6. Boldyreff B, Meggio F, Pinna LA, Issinger O-G: Reconstitution of normal and hyperactivated forms of casein kinase-2 by variably mutated β-subunits. Biochemistry 32: 12672–12677 (1993).

    Google Scholar 

  7. Boldyreff B, Piontek K, Schmidt-Spaniol I, Issinger O-G: The β subunit of casein kinase II: cloning of cDNAs from murine and porcine orgin and expression of the porcine sequence as a fusion protein. Biochim Biophys Acta 1088: 439–441 (1991).

    Google Scholar 

  8. Chen-Wu J, Padmanabha R, Glover CVC: Isolation, sequencing, and disruption of the CKA1 gene encoding the alpha subunit of yeast casein kinase II. Mol Cell Biol 8: 4981–4990 (1988).

    Google Scholar 

  9. Dobrowolska G, Boldryeff B, Issinger O-G: Cloning and sequencing of the casein kinase 2 α subunit from Zea mays. Biochim Biophys Acta 1129: 139–140 (1991).

    Google Scholar 

  10. Dobrowolska G, Meggio F, Pinna LA: Characterization of multiple forms of maize seedling protein kinases reminiscent of animal casein kinases S (type 1) and TS (type 2). Biochim Biophys Acta 931: 188–195 (1987).

    Google Scholar 

  11. Dobrowolska G, Meggio F, Szczegielnaik J, Muszynska G, Pinna LA. Purification and characterization of maize seedling casein kinase IIB, a monomeric enzyme immunologically related to the α subunit of animal casein kinase-2. Eur J Biochem 204: 299–303 (1992).

    Google Scholar 

  12. Dower WJ, Miller JF, Ragsdale CW: High efficiency transformation of E. coli by high voltage electroporation. Nucl Acids Res 16: 6127–6145 (1988).

    Google Scholar 

  13. Elble R. A simple and efficient procedure for transformation of yeasts. Bio Techniques 13: 18–20 (1992).

    Google Scholar 

  14. Gaber RF, Culbertson MR. Frameshift suppression in Saccharomyces cerevisiae. IV. New suppressors among spontaneous co-revertants of the group II his4–206 and LEU2–3 frameshift mutations. Genetics 101: 345–367 (1982).

    Google Scholar 

  15. Gowda S, Pillay DTN: Cyclic AMP independent protein kinases from soybean cotyledons (Glycine max. L.). Plant Sci Lett 25: 49–59 (1982).

    Google Scholar 

  16. Haffner MH, Chin MB, Lane BG: Wheat embryo ribonucleates. XII. Formal characterization of terminal and penultimate nucleoside residues at the 5′ ends of capped RNA from imbibing wheat embryos. Can J Biochem 56: 729–733 (1978).

    Google Scholar 

  17. Hoffman CS, Winston F: A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57: 267–272 (1987).

    Google Scholar 

  18. Hu E, Rubin CS: Casein kinase II from Caenorhabditis elegans: cloning, characterization, and developmental regulation of the gene encoding the β subunit. J Biol Chem 266: 19796–19802 (1991).

    Google Scholar 

  19. Jakobi R, Voss H, Pyerin W: Human phosvitin/casein kinase type II. Eur J Biochem 183: 227–233 (1989).

    Google Scholar 

  20. Jedlicki A, Hinrichs MV, Allende CC, Allende JE: The cDNAs coding for the α- and β-subunits of Xenopus laevis casein kinase II. FEBS Lett 297: 280–284 (1992).

    Google Scholar 

  21. Johnston M, Davis RW: Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol 4: 1440–1448 (1984).

    Google Scholar 

  22. Kassir Y, Simchen G. Monitoring meiosis and sporulation in Saccharomyces cerevisiae. Meth Enzymol 194: 94–110 (1991).

    Google Scholar 

  23. Klimczak LJ, Schindler U, Cashmore AR: DNA binding activity of the Arabidopsis G-box binding factor GBF1 is stimulated by phosphorylation by casein kinase II from broccoli. Plant Cell 4: 87–98 (1992).

    Google Scholar 

  24. Li H, Roux SJ: Casein kinase II protein kinase is bound to lamina-matrix and phosphorylates lamin-like protein in isolated pea nuclei. Proc Natl Acad Sci USA 89: 8434–8438 (1992).

    Google Scholar 

  25. Li H, Roux SJ: Purification and characterization of casein kinase 2-type protein kinase from pea nuclei. Plant Physiol 99: 686–692 (1992).

    Google Scholar 

  26. Lin W-J, Tuazon PT, Traugh JA: Characterization of the catalytic subunit of casein kinase II expressed in Escherichia coli and regulation of activity. J Biol Chem 266: 5664–5669 (1991).

    Google Scholar 

  27. Litchfield SW, Lozeman FJ, Cicirelli MF, Harrylock M, Ericsson LH, Piening CJ, Krebs EG. Phosphorylation of the β subunit of casein kinase II in human A431 cells. J Biol Chem 266: 20380–20389 (1991).

    Google Scholar 

  28. Livak KJ: Detailed structure of the Drosophila melanogaster stellate genes and their transcripts. Genetics 124: 303–316 (1990).

    Google Scholar 

  29. Meggio F, Boldryeff B, Issinger O-G, Pinna LA: The autophosphorylation and p34cdc2 phosphorylation sites of casein kinase-2 β-subunit are not essential for reconstituting the fully-active heterotetrameric holoenzyme. Biochim Biophys Acta 1164: 223–225 (1993).

    Google Scholar 

  30. Meggio F, Boldryeff B, Marin O, Pinna LA, Issinger O-G: Role of the β subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted enzyme. Eur J Biochem 204: 293–297 (1992).

    Google Scholar 

  31. Mizoguchi T, Yamaguchi-Shinozaki K, Hayashida N, Kamada H, Shinozaki K: Cloning and characterization of two cDNAs encoding casein kinase II catalytic subunits in Arabidopsis thaliana. Plant Mol Biol 21: 279–289 (1993).

    Google Scholar 

  32. Padmanabha R, Chen-Wu J, Hanna DE, Glover CVC: Isolation, sequencing, and disruption of the yeast CKA2 gene: casein kinase II is essential for viability in Saccharomyces cerevisiae. Mol Cell Biol 10: 489–4099 (1990).

    Google Scholar 

  33. Padmanabha R, Glover CVC: Casein kinase II of yeast contains two distinct α polypeptides and an unusually large β subunit. J Biol Chem 262: 1829–1835 (1987).

    Google Scholar 

  34. Pinna LA: Casein kinase 2: an ‘eminence grise’ in cellular regulation? Biochim Biophys Acta 1054: 267–284 (1990).

    Google Scholar 

  35. Plana M, Itarte E, Eritja R, Goday A, Pages M, Martinez MC: Phosphorylation of maize RAB-17 protein by casein kinase 2. J Biol Chem 266: 22510–22514 (1991).

    Google Scholar 

  36. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  37. Sanger F, Miklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 4: 5463–5467 (1977).

    Google Scholar 

  38. Saxena A, Padmanabha R, Glover CVC: Isolation and sequencing of cDNA clones encoding alpha and beta subunits of Drosophila melanogaster casein kinase II. Mol Cell Biol 7: 3409–3417 (1987).

    Google Scholar 

  39. Schiestl RH, Gietz RD: High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16: 339–346 (1989).

    Google Scholar 

  40. Short JM, Fernandez JM, Sorge JA, Huse WD: λZAP: A bacteriophage λ expression vector with in vivo excision properties. Nucl Acids Res 16: 7583 (1988).

    Google Scholar 

  41. Sikorski R, Hieter P: A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19–27 (1989).

    Google Scholar 

  42. Smith RD, Walker JC: Expression of multiple type I phosphoprotein phosphatases in Arabidopsis thalic Plant Mol Biol 21: 307–316 (1993).

    Google Scholar 

  43. Tuazon PT, Traugh JA: Casein kinase I and II—multipotential serine protein kinases: structure, function and regulation. In: Greengard P, Robinson GA (eds) Advances in Second Messenger and Phosphoprotein Research, pp. 123–164. Raven Press, New York (1991).

    Google Scholar 

  44. Yan T-FJ, Tao M: Purification and characterization of wheat germ protein kinase. J Biol Chem 257: 7037–7043 (1982).

    Google Scholar 

  45. Zhang S, Jin C-D, Roux SJ: Casein kinase II-type protein kinase from pea cytoplasm and its inactivation by alkaline phospahtase in vitro. Plant Physiol 103: 955–962 (1993).

    Google Scholar 

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Collinge, M.A., Walker, J.C. Isolation of an Arabidopsis thaliana casein kinase II β subunit by complementation in Saccharomyces cerevisiae . Plant Mol Biol 25, 649–658 (1994). https://doi.org/10.1007/BF00029603

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