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A specific catalytic subunit isoform of protein kinase CK2 is required for phosphorylation of the repressor Nrg1 in Saccharomyces cerevisiae

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Abstract

Protein kinase CK2 is highly conserved in eukaryotes and plays roles in many different cellular processes. CK2 is a tetramer comprising two catalytic and two regulatory subunits. Most organisms have two major isoforms of the catalytic subunit, and evidence suggests strongly overlapping function. In the yeast Saccharomyces cerevisiae, CK2 is essential for viability, and either catalytic subunit isoform, Cka1 or Cka2, suffices, but previous genetic evidence suggests that the isoforms have some distinct roles. In this work, we present evidence that the transcriptional repressor Nrg1, which regulates various stress-responsive genes, is a downstream target of CK2 containing the Cka1 isoform. We found that Nrg1 is phosphorylated in response to stress and that its phosphorylation was defective in cka1Δ, but not cka2Δ, mutants. Thus, the Cka1 catalytic subunit isoform is specifically required for phosphorylation of Nrg1 in vivo. The CK2 regulatory subunits were also required, indicating that the CK2 holoenzyme is involved. Both yeast and recombinant human CK2 phosphorylated recombinant Nrg1 in vitro. This identification of a protein whose phosphorylation requires a specific CK2 catalytic subunit isoform supports the view that the two isoforms exhibit functional specificity in vivo.

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References

  • Ackermann K, Waxmann A, Glover CV, Pyerin W (2001) Genes targeted by protein kinase CK2: a genome-wide expression array analysis in yeast. Mol Cell Biochem 227:59–66

    Article  PubMed  CAS  Google Scholar 

  • Ahmed K, Gerber DA, Cochet C (2002) Joining the cell survival squad: an emerging role for protein kinase CK2. Trends Cell Biol 12:226–230

    Article  PubMed  CAS  Google Scholar 

  • Allende JE, Allende CC (1995) Protein kinases. 4. Protein kinase CK2: an enzyme with multiple substrates and a puzzling regulation. FASEB J 9:313–323

    PubMed  CAS  Google Scholar 

  • Berkey CD, Vyas VK, Carlson M (2004) Nrg1 and Nrg2 transcriptional repressors are differently regulated in response to carbon source. Eukaryot Cell 3:311–317

    Article  PubMed  CAS  Google Scholar 

  • Bidwai AP, Reed JC, Glover CV (1995) Cloning and disruption of CKB1, the gene encoding the 38-kDa beta subunit of Saccharomyces cerevisiae casein kinase II (CKII). Deletion of CKII regulatory subunits elicits a salt-sensitive phenotype. J Biol Chem 270:10395–10404

    Article  PubMed  CAS  Google Scholar 

  • Boldyreff B, Mietens U, Issinger OG (1996) Structure of protein kinase CK2: dimerization of the human beta-subunit. FEBS Lett 379:153–156

    Article  PubMed  CAS  Google Scholar 

  • Bosc DG, Slominski E, Sichler C, Litchfield DW (1995) Phosphorylation of casein kinase II by p34cdc2. Identification of phosphorylation sites using phosphorylation site mutants in vitro. J Biol Chem 270:25872–25878

    Article  PubMed  CAS  Google Scholar 

  • Bruno VM, Mitchell AP (2005) Regulation of azole drug susceptibility by Candida albicans protein kinase CK2. Mol Microbiol 56:559–573

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Domanska K, et al (2005) Different properties of four molecular forms of protein kinase CK2 from Saccharomyces cerevisiae. Acta Biochim Pol 52:947–951

    PubMed  CAS  Google Scholar 

  • Elbing K, McCartney RR, Schmidt MC (2006) Purification and characterization of the three Snf1-activating kinases of Saccharomyces cerevisiae. Biochem J 393:797–805

    Article  PubMed  CAS  Google Scholar 

  • Gavin AC, et al (2002) Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature 415:141–147

    Article  PubMed  CAS  Google Scholar 

  • Gietz RD, Graham KC, Litchfield DW (1995) Interactions between the subunits of casein kinase II. J Biol Chem 270:13017–13021

    Article  PubMed  CAS  Google Scholar 

  • Glover CV III (1998) On the physiological role of casein kinase II in Saccharomyces cerevisiae. Prog Nucleic Acid Res Mol Biol 59:95–133

    Article  PubMed  CAS  Google Scholar 

  • Guarente L (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Meth Enzymol 101:181–191

    Article  PubMed  CAS  Google Scholar 

  • Hanna DE, Rethinaswamy A, Glover CV (1995) Casein kinase II is required for cell cycle progression during G1 and G2/M in Saccharomyces cerevisiae. J Biol Chem 270:25905–25914

    Article  PubMed  CAS  Google Scholar 

  • Heriche JK, Lebrin F, Rabilloud T, Leroy D, Chambaz EM, Goldberg Y (1997) Regulation of protein phosphatase 2A by direct interaction with casein kinase 2alpha. Science 276:952–955

    Article  PubMed  CAS  Google Scholar 

  • Kartasheva NN, Kuchin SV, Benevolensky SV (1996) Genetic aspects of carbon catabolite repression of the STA2 glucoamylase gene in Saccharomyces cerevisiae. Yeast 12:1297–1300

    Article  PubMed  CAS  Google Scholar 

  • Kuchin S, Vyas VK, Carlson M (2002) Snf1 protein kinase and the repressors Nrg1 and Nrg2 regulate FLO11, haploid invasive growth, and diploid pseudohyphal differentiation. Mol Cell Biol 22:3994–4000

    Article  PubMed  CAS  Google Scholar 

  • Lamb TM, Mitchell AP (2003) The transcription factor Rim101p governs ion tolerance and cell differentiation by direct repression of the regulatory genes NRG1 and SMP1 in Saccharomyces cerevisiae. Mol Cell Biol 23:677–686

    Article  PubMed  CAS  Google Scholar 

  • Litchfield DW (2003) Protein kinase CK2: structure, regulation and role in cellular decisions of life and death. Biochem J 369:1–15

    Article  PubMed  CAS  Google Scholar 

  • Martzen MR, et al (1999) A biochemical genomics approach for identifying genes by the activity of their products. Science 286:1153–1155

    Article  PubMed  CAS  Google Scholar 

  • Meggio F, Pinna LA (2003) One-thousand-and-one substrates of protein kinase CK2? FASEB J 17:349–368

    Article  PubMed  CAS  Google Scholar 

  • Messenger MM, Saulnier RB, Gilchrist AD, Diamond P, Gorbsky GJ, Litchfield DW (2002) Interactions between protein kinase CK2 and Pin1. Evidence for phosphorylation-dependent interactions. J Biol Chem 277:23054–23064

    Article  PubMed  CAS  Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Niefind K, Guerra B, Ermakowa I, Issinger OG (2001) Crystal structure of human protein kinase CK2: insights into basic properties of the CK2 holoenzyme. EMBO J 20:5320–5331

    Article  PubMed  CAS  Google Scholar 

  • Olsten ME, Litchfield DW (2004) Order or chaos? An evaluation of the regulation of protein kinase CK2. Biochem Cell Biol 82:681–693

    Article  PubMed  CAS  Google Scholar 

  • Padmanabha R, Chen-Wu JL, Hanna DE, Glover CV (1990) Isolation, sequencing, and disruption of the yeast CKA2 gene: casein kinase II is essential for viability in Saccharomyces cerevisiae. Mol Cell Biol 10:4089–4099

    PubMed  CAS  Google Scholar 

  • Park SH, Koh SS, Chun JH, Hwang HJ, Kang HS (1999) Nrg1 is a transcriptional repressor for glucose repression of STA1 gene expression in Saccharomyces cerevisiae. Mol Cell Biol 19:2044–2050

    PubMed  CAS  Google Scholar 

  • Pinna LA (2002) Protein kinase CK2: a challenge to canons. J Cell Sci 115:3873–3878

    Article  PubMed  CAS  Google Scholar 

  • Rethinaswamy A, Birnbaum MJ, Glover CV (1998) Temperature-sensitive mutations of the CKA1 gene reveal a role for casein kinase II in maintenance of cell polarity in Saccharomyces cerevisiae. J Biol Chem 273:5869–5877

    Article  PubMed  CAS  Google Scholar 

  • Roberts RL, Fink GR (1994) Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth. Genes Dev 8:2974–2985

    Article  PubMed  CAS  Google Scholar 

  • Rose MD, Winston F, Hieter P (1990) Methods in yeast genetics: a laboratory course manual. Cold Spring Harbor Laboratory Press, Plainview, New York

    Google Scholar 

  • Rothfels K, Tanny JC, Molnar E, Friesen H, Commisso C, Segall J (2005) Components of the ESCRT pathway, DFG16, and YGR122w are required for Rim101 To Act as a corepressor with Nrg1 at the negative regulatory element of the DIT1 Gene of Saccharomyces cerevisiae. Mol Cell Biol 25:6772–6788

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Song W, Carlson M (1998) Srb/Mediator proteins interact functionally and physically with transcriptional repressor Sfl1. EMBO J 17:5757–5765

    Article  PubMed  CAS  Google Scholar 

  • Thomas BJ, Rothstein R (1989) Elevated recombination rates in transcriptionally active DNA. Cell 56:619–630

    Article  PubMed  CAS  Google Scholar 

  • Vilk G, Saulnier RB, St Pierre R, Litchfield DW (1999) Inducible expression of protein kinase CK2 in mammalian cells. Evidence for functional specialization of CK2 isoforms. J Biol Chem 274:14406–14414

    Article  PubMed  CAS  Google Scholar 

  • Vincent O, Carlson M (1999) Gal83 mediates the interaction of the Snf1 kinase complex with the transcription activator Sip4. EMBO J 18:6672–6681

    Article  PubMed  CAS  Google Scholar 

  • Vyas VK, Berkey CD, Miyao T, Carlson M (2005) Repressors Nrg1 and Nrg2 regulate a set of stress-responsive genes in Saccharomyces cerevisiae. Eukaryot Cell 4:1882–1891

    Article  PubMed  CAS  Google Scholar 

  • Vyas VK, Kuchin S, Berkey CD, Carlson M (2003) Snf1 kinases with different β-subunit isoforms play distinct roles in regulating haploid invasive growth. Mol Cell Biol 23:1341–1348

    Article  PubMed  CAS  Google Scholar 

  • Vyas VK, Kuchin S, Carlson M (2001) Interaction of the repressors Nrg1 and Nrg2 with the Snf1 protein kinase in Saccharomyces cerevisiae. Genetics 158:563–572

    PubMed  CAS  Google Scholar 

  • Xu X, Toselli PA, Russell LD, Seldin DC (1999) Globozoospermia in mice lacking the casein kinase II alpha’ catalytic subunit. Nat Genet 23:118–121

    Article  PubMed  CAS  Google Scholar 

  • Yu IJ, Spector DL, Bae YS, Marshak DR (1991) Immunocytochemical localization of casein kinase II during interphase and mitosis. J Cell Biol 114:1217–1232

    Article  PubMed  CAS  Google Scholar 

  • Zhou H, Winston F (2001) NRG1 is required for glucose repression of the SUC2 and GAL genes of Saccharomyces cerevisiae. BMC Genet 2:5

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Valmik Vyas for plasmids and advice and Vinny Bruno for discussion. This work was supported by National Institutes of Health grant GM34095 to M.C.

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Correspondence to Marian Carlson.

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Communicated by J. Heitman

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Berkey, C.D., Carlson, M. A specific catalytic subunit isoform of protein kinase CK2 is required for phosphorylation of the repressor Nrg1 in Saccharomyces cerevisiae . Curr Genet 50, 1–10 (2006). https://doi.org/10.1007/s00294-006-0070-5

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