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Abstract:

We have previously reported that Candida albicans protein kinase CK2 is composed of two distinct catalytic (α- and α′-) and two distinct regulatory (β- and β′-) subunits. We report here the isolation of two cDNAs clones, CaCKB1 and CaCKB2, encoding C. albicans β- and β′-subunits, respectively. The predicted β- and β′- proteins have calculated molecular masses of 34 kDa and 31 kDa and show all major features conserved in β-subunits of other organisms, including the N-terminal autophosphorylation site, the internal acidic region and a potential metal-binding motif. The deduced amino acid sequence of C. albicans β-subunit displays 48% identity with that of Saccharomyces cerevisiae and has an unusually long C-terminal acidic region containing a putative autophosphorylation site. C. albicans β′ shows 54% sequence identity with its S. cerevisiae homologue. Semi-quantitative RT-PCR analyses indicate that the mRNAs corresponding to both subunits are present in similar amounts in the yeast and hyphal forms of the fungus. To evaluate the biochemical properties of C. albicans β- and β′-subunits, both proteins were expressed in Escherichia coli and purified. Experiments performed in vitro indicate that both recombinant subunits reconstitute a fully functional holoenzyme when incubated with stoichiometric amounts of human recombinant α-subunit, as judged by their ability to abolish basal phosphorylation of calmodulin by human recombinant α-subunit and the reversion of the inhibitory effect by polylysine. In addition, both regulatory subunits can be phosphorylated by human recombinant α subunit. Phylogenetic analysis of β- and β′-proteins of C. albicans and other organisms shows that the CKB gene duplication occurred before the split of the ascomycete and basidiomycete lineages. cDNA sequences of C. albicans CKB1 (Accession No. AF0599060) and CKB2 (Accession No. AY172319) have been deposited in the GenBank database. Copyright © 2003 John Wiley & Sons, Ltd.

Registro:

Documento: Artículo
Título:cDNA cloning, biochemical and phylogenetic characterization of β- and β' -subunits of Candida albicans protein kinase CK2
Autor:Zelada, A.; De Souza, F.S.J.; Walz, K.; Giasson, L.; Passeron, S.
Filiación:Catedra de Microbiología, Facultad de Agronomía, Universidad de Buenos Aires, Avda. San Martín 4453, 1417 Buenos Aires, Argentina
School of Dentistry, GREB, Laval University, Sainte-Foy, Que. G1K 7P4, Canada
INGEBI-CONICET, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina
Department of Human Genetics, Baylor College of Medicine, 604 B One Baylor Plaza, Houston, TX 77030, United States
Palabras clave:Candida albicans; CK2; Phylogeny; Protein kinase; Regulatory subunit; calmodulin; casein kinase II; complementary DNA; holoenzyme; messenger RNA; polylysine; protein subunit; recombinant protein; alpha chain; amino acid sequence; amino terminal sequence; article; Ascomycetes; autophosphorylation; Basidiomycetes; beta chain; calculation; Candida albicans; carboxy terminal sequence; controlled study; gene duplication; genetic conservation; in vitro study; incubation time; metal binding; molecular cloning; molecular weight; nonhuman; nucleotide sequence; phylogeny; prediction; priority journal; promoter region; protein analysis; protein expression; protein function; protein isolation; protein motif; protein phosphorylation; protein purification; reverse transcription polymerase chain reaction; Saccharomyces cerevisiae; sequence homology; stoichiometry; Amino Acid Sequence; Binding Sites; Candida albicans; Casein Kinase II; Cloning, Molecular; Dimerization; DNA Primers; Kinetics; Molecular Sequence Data; Phosphorylation; Phylogeny; Protein Subunits; Protein-Serine-Threonine Kinases; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Sequence Homology, Amino Acid; Zinc Fingers; Candida; Candida albicans; Escherichia coli; Fungi; Saccharomyces; Saccharomyces cerevisiae
Año:2003
Volumen:20
Número:6
Página de inicio:471
Página de fin:478
DOI: http://dx.doi.org/10.1002/yea.977
Título revista:Yeast
Título revista abreviado:Yeast
ISSN:0749503X
CODEN:YESTE
CAS:polylysine, 25104-18-1, 25988-63-0, 33960-24-6, 38000-06-5, 73565-56-7; Casein Kinase II, EC 2.7.1.37; DNA Primers; Protein Subunits; Protein-Serine-Threonine Kinases, EC 2.7.1.37; Recombinant Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0749503X_v20_n6_p471_Zelada

Referencias:

  • Allende, J.E., Allende, C.C., Protein kinase CK2: An enzyme with multiple substrates and a puzzling regulation (1995) FASEB J., 9, pp. 313-323
  • Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., Basic local alignment search tool (1990) J. Mol. Biol., 215, pp. 403-410
  • Ausubel, F.M., Brent, R., Kingston, R.E., (1994) Current Protocols in Molecular Biology, , Chanda VB (coordinating ed.). Wiley: New York
  • Bidwai, A.P., Reed, J.C., Glover, C.V., Cloning and disruption of CKB1, the gene encoding the 38 kDa subunit of Saccharomyces cerevisiae casein kinase II (CKII). Deletion of CKII regulatory subunits elicits a salt-sensitive phenotype (1995) J. Biol. Chem., 270, pp. 10395-10404
  • Chantalat, L., Leroy, D., Filhol, O., Crystal structure of the human protein kinase CK2 regulatory subunit reveals its zinc finger-mediated dimerization (1999) EMBO J., 18, pp. 2930-2940
  • Chen-Wu, J.L.P., Padmanabha, R., Glover, C.V.C., Isolation, sequencing, and disruption of the CKA1 gene encoding the α-subunit of yeast casein kinase II (1988) Mol. Cell Biol., 8, pp. 4981-4990
  • Felsenstein, J., (1995) PHYLIP (Phylogeny Inference Package), , Version 3.57c, University of Washington, Seattle
  • Glover C.V. III, On the physiological role of casein kinase II in Saccharomyces cerevisiae (1998) Prog. Nucleic Acid. Res. Mol. Biol., 59, pp. 95-133
  • Isola, N.R., Harn, H.J., Cooper, D.L., Screening recombinant DNA libraries: A rapid and efficient method for isolating cDNA clones utilizing the PCR (1991) Biotechniques, 11, pp. 580-582
  • Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685
  • Leroy, D., Heriche, J.K., Filhol, O., Chambaz, E.M., Cochet, C., Binding of polyamines to an autonomous domain of the regulatory subunit of protein kinase CK2 induces a conformational change in the holoenzyme. A proposed role for the kinase stimulation (1995) J. Biol. Chem., 272, pp. 820-827
  • Meggio, F., Boldyreff, B., Marin, O., Pinna, L., Issinger, O.-G., Role of β-subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted enzyme (1992) Eur. J. Biochem., 204, pp. 293-297
  • Meggio, F., Boldyreff, B., Marin, O., The effect of polylysine on casein-kinase-2 activity is influenced by both the structure of the protein/peptide substrates and the subunit composition of the enzyme (1992) Eur. J. Biochem., 205, pp. 939-945
  • Meggio, F., Boldyreff, B., Issinger, O.-G., Pinna, L.A., Casein kinase 2 downregulation and activation by polybasic peptides are mediated by acidic residues in the 55-64 region of the β-subunit. A study with calmodulin as phosphorylatable substrate (1994) Biochemistry, 33, pp. 4336-4342
  • Padmanabha, R., Chen-Wu, J.L.P., Hanna, D.E., Glover, C.V.C., Isolation, sequencing, and disruption of the yeast CKA2 gene: Casein kinase II is essential for viability in Saccharomyces cerevisiae (1990) Mol. Cell Biol., 10, pp. 4089-4099
  • Pinna, L., Protein kinase CK2 (1997) Int. J. Biochem. Cell Biol., 29, pp. 551-554
  • Reed, J.C., Bidwai, A.P., Glover, C.V., Cloning and disruption of CKB2, the gene encoding the 32 kDa regulatory β′-subunit of Saccharomyces cerevisiae casein kinase II (1994) J. Biol. Chem., 269, pp. 18192-18200
  • Roussou, I., Draetta, G., The Schizosaccharomyces pombe casein kinase II α- and β-subunits: Evolutionary conservation and positive role of the β-subunit (1994) Mol. Cell Biol., 14, pp. 576-586
  • Saitou, N., Nei, M., The neighbor-joining method: A new method for reconstructing phylogenetic trees (1987) Mol. Biol. Evol., 6, pp. 514-525
  • Sambrook, J., Russel, D.W., (2000) Molecular Cloning: A Laboratory Manual, , Cold Spring Harbor Laboratory Press: New York
  • Sanger, F., Nicklen, S., Coulson, A.R., DNA sequencing with chain-terminating inhibitors (1977) Proc. Natl. Acad. Sci. USA, 74, pp. 5463-5467
  • Sarkar, G., Sommer, S.S., The 'megaprimer' method of site-directed mutagenesis (1990) Biotechniques, 8, pp. 404-407
  • Shepherd, M.G., Yin, C.Y., Ram, S.P., Sullivan, P.A., Germ tube induction in Candida albicans (1980) Can. J. Microbiol., 26, pp. 21-26
  • Souciet, J., Aigle, M., Artiguenave, F., Genomic exploration of' the hemiascomycetous yeasts: 1. A set of yeasts species for molecular evolution studies (2000) FEBS Lett., 487, pp. 3-12
  • Sugano, S., Andronis, C., Green, R.M., Wang, Z.Y., Tobin, E.M., Protein kinase CK2 interacts with and phosphorylates the Arabidopsis circadian clock-associated 1 protein (1998) Proc. Natl. Acad. Sci. USA, 95, pp. 11020-11025
  • Swoboda, R.K., Bertram, G., Hollander, H., Glycolytic enzymes of Candida albicans are non-ubiquitous immunogen during candidiasis (1993) Infect. Human, 61, pp. 4263-4271
  • Thompson, J.D., Higgins, D.G., Gibson, T.J., CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice (1994) Nucleic Acids Res., 22, pp. 4673-4680
  • Walz, K., Pardo, P., Passeron, S., Purification and characterization of protein kinase CK2 from Candida albicans: Evidence for the presence of two distinct regulatory subunits, β and β′ (1997) Arch. Biochem. Biophys., 340, pp. 347-354
  • Wong, S., Butler, G., Wolfe, K.H., Gene order evolution and paleopolyploidy in hemiascomycete yeasts (2002) Proc. Natl. Acad. Sci. USA, 99, pp. 9272-9277
  • Wood, V., Gwilliam, R., Rajandream, M.A., The genome sequence of Schizosaccharomyces pombe (2002) Nature, 415, pp. 871-880
  • Yang, Y., Cheng, P., Liu, Y., Regulation of the Neurospora circadian clock by casein kinase II (2002) Genes Dev., 16, pp. 994-1006

Citas:

---------- APA ----------
Zelada, A., De Souza, F.S.J., Walz, K., Giasson, L. & Passeron, S. (2003) . cDNA cloning, biochemical and phylogenetic characterization of β- and β' -subunits of Candida albicans protein kinase CK2. Yeast, 20(6), 471-478.
http://dx.doi.org/10.1002/yea.977
---------- CHICAGO ----------
Zelada, A., De Souza, F.S.J., Walz, K., Giasson, L., Passeron, S. "cDNA cloning, biochemical and phylogenetic characterization of β- and β' -subunits of Candida albicans protein kinase CK2" . Yeast 20, no. 6 (2003) : 471-478.
http://dx.doi.org/10.1002/yea.977
---------- MLA ----------
Zelada, A., De Souza, F.S.J., Walz, K., Giasson, L., Passeron, S. "cDNA cloning, biochemical and phylogenetic characterization of β- and β' -subunits of Candida albicans protein kinase CK2" . Yeast, vol. 20, no. 6, 2003, pp. 471-478.
http://dx.doi.org/10.1002/yea.977
---------- VANCOUVER ----------
Zelada, A., De Souza, F.S.J., Walz, K., Giasson, L., Passeron, S. cDNA cloning, biochemical and phylogenetic characterization of β- and β' -subunits of Candida albicans protein kinase CK2. Yeast. 2003;20(6):471-478.
http://dx.doi.org/10.1002/yea.977