PCR cloning and sequence of two cDNAs encoding the alpha and beta subunits of rabbit casein kinase-II

Abstract We have recently shown that inhibition of protein phosphatases in platelets causes increases in protein phosphorylations with a concomitant inhibition of platelet responses. The burst in protein phosphorylation appears to be catalyzed by messenger-independent protein kinases. The aim of the present study was to characterize the presence of broad families of protein kinases found in platelets. Lysates of control and thrombin-stimulated platelets were prepared, and proteins were separated on MONO Q fast protein liquid chromatography. In addition to the presence of histone protein kinase and tyrosine kinase activities, human platelets contain casein kinase II (CKII) activity as assessed by phosphorylation of a specific substrate peptide. Western blot analysis and immunogold electron microscopy studies further showed the presence of alpha-, alpha'-, and beta- subunits of CKII. The enzyme appears to be distributed throughout the cytosol and not secreted after thrombin treatment. Immunoprecipitation studies suggest that at least some of the holoenzymes exist as an alpha alpha' beta 2 complex. Although no activation of the enzyme was detected after thrombin treatment, our results show that CKII is a major messenger-independent protein kinase in platelets.

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Isolation, sequencing, and disruption of the yeast CKA2 gene: casein kinase II is essential for viability in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.10.8.4089-4099.1990 ◽

1990 ◽

Vol 10 (8) ◽

pp. 4089-4099 ◽

Cited By ~ 1

Author(s):

R Padmanabha ◽

J L Chen-Wu ◽

D E Hanna ◽

C V Glover

Keyword(s):

Saccharomyces Cerevisiae ◽

Growth Arrest ◽

Casein Kinase Ii ◽

Gene Replacement ◽

Casein Kinase ◽

Alpha Subunit ◽

Beta Subunits ◽

Catalytic Subunits ◽

Yeast Strains ◽

Cell Biol

Casein kinase II of Saccharomyces cerevisiae contains two distinct catalytic subunits, alpha and alpha', which are encoded by the CKA1 and CKA2 genes, respectively. Null mutations in the CKA1 gene do not confer a detectable phenotype (J. L.-P. Chen-Wu, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 8:4981-4990, 1988), presumably because of the presence of the CKA2 gene. We report here the cloning, sequencing, and disruption of the CKA2 gene. The alpha' subunit encoded by the CKA2 gene is 60% identical to the CKA1-encoded alpha subunit and 55% identical to the Drosophila alpha subunit (A. Saxena, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 7:3409-3417, 1987). Deletions of the CKA2 gene were constructed by gene replacement techniques. Haploid cells in which the CKA2 gene alone is disrupted show no detectable phenotype, but haploid cells carrying disruptions in both the CKA1 and CKA2 genes are inviable. Cells in which casein kinase II activity is depleted increase substantially in size prior to growth arrest, and a significant fraction of the arrested cells exhibit a pseudomycelial morphology. Disruption of the activity also results in flocculation. Yeast strains lacking both endogenous catalytic subunit genes can be rescued by expression of the alpha and beta subunits of Drosophila casein kinase II or by expression of the Drosophila alpha subunit alone, suggesting that casein kinase II function has been conserved through evolution.

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The Schizosaccharomyces pombe casein kinase II alpha and beta subunits: evolutionary conservation and positive role of the beta subunit.

Molecular and Cellular Biology ◽

10.1128/mcb.14.1.576 ◽

1994 ◽

Vol 14 (1) ◽

pp. 576-586 ◽

Cited By ~ 72

Author(s):

I Roussou ◽

G Draetta

Keyword(s):

Enzyme Activity ◽

Amino Acid ◽

Amino Acid Sequence ◽

Schizosaccharomyces Pombe ◽

Sequence Similarity ◽

Casein Kinase Ii ◽

Casein Kinase ◽

Beta Subunit ◽

Beta Subunits ◽

High Sequence Similarity

Casein kinase II is a key regulatory enzyme involved in many cellular processes, including the control of growth and cell division. We report the molecular cloning and sequencing of cDNAs encoding the alpha and the beta subunits of casein kinase II of Schizosaccharomyces pombe. The deduced amino acid sequence of Cka1, the alpha catalytic subunit, shows high sequence similarity to alpha subunits identified in other species. The amino acid sequence of Ckb1, the S. pombe beta subunit, is 57% identical to that of the human beta subunit. Cka1 overexpression results in no detectable phenotype. In contrast, Ckb1 overexpression inhibits cell growth and cytokinesis, with formation of multiseptated cells. Disruption of the ckb1+ gene causes a cold-sensitive phenotype and abnormalities in cell shape. In these cells, the casein kinase II activity is reduced to undetectable levels, demonstrating that Ckb1 is required for enzyme activity in vivo. In agreement with this, the activity measured in a strain expressing high levels of Cka1 is enhanced only when the Ckb1 protein is coexpressed. Altogether, our data suggest that Ckb1 is a positive regulator of the enzyme activity, and that it plays a role in mediating the interaction of casein kinase II with downstream targets and/or with additional regulators.

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Casein kinase II. cDNA sequences, developmental expression, and tissue distribution of mRNAs for alpha, alpha', and beta subunits of the chicken enzyme

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)52252-3 ◽

1991 ◽

Vol 266 (4) ◽

pp. 2362-2368 ◽

Cited By ~ 4

Author(s):

G Maridor ◽

W Park ◽

W Krek ◽

E A Nigg

Keyword(s):

Tissue Distribution ◽

Casein Kinase Ii ◽

Casein Kinase ◽

Developmental Expression ◽

Beta Subunits ◽

Cdna Sequences

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Isolation, sequencing, and disruption of the yeast CKA2 gene: casein kinase II is essential for viability in Saccharomyces cerevisiae.

Molecular and Cellular Biology ◽

10.1128/mcb.10.8.4089 ◽

1990 ◽

Vol 10 (8) ◽

pp. 4089-4099 ◽

Cited By ~ 240

Author(s):

R Padmanabha ◽

J L Chen-Wu ◽

D E Hanna ◽

C V Glover

Keyword(s):

Saccharomyces Cerevisiae ◽

Growth Arrest ◽

Casein Kinase Ii ◽

Gene Replacement ◽

Casein Kinase ◽

Alpha Subunit ◽

Beta Subunits ◽

Catalytic Subunits ◽

Yeast Strains ◽

Cell Biol

Casein kinase II of Saccharomyces cerevisiae contains two distinct catalytic subunits, alpha and alpha', which are encoded by the CKA1 and CKA2 genes, respectively. Null mutations in the CKA1 gene do not confer a detectable phenotype (J. L.-P. Chen-Wu, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 8:4981-4990, 1988), presumably because of the presence of the CKA2 gene. We report here the cloning, sequencing, and disruption of the CKA2 gene. The alpha' subunit encoded by the CKA2 gene is 60% identical to the CKA1-encoded alpha subunit and 55% identical to the Drosophila alpha subunit (A. Saxena, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 7:3409-3417, 1987). Deletions of the CKA2 gene were constructed by gene replacement techniques. Haploid cells in which the CKA2 gene alone is disrupted show no detectable phenotype, but haploid cells carrying disruptions in both the CKA1 and CKA2 genes are inviable. Cells in which casein kinase II activity is depleted increase substantially in size prior to growth arrest, and a significant fraction of the arrested cells exhibit a pseudomycelial morphology. Disruption of the activity also results in flocculation. Yeast strains lacking both endogenous catalytic subunit genes can be rescued by expression of the alpha and beta subunits of Drosophila casein kinase II or by expression of the Drosophila alpha subunit alone, suggesting that casein kinase II function has been conserved through evolution.

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Identifying and characterizing casein kinase II in human platelets

Blood ◽

10.1182/blood.v83.12.3517.bloodjournal83123517 ◽

1994 ◽

Vol 83 (12) ◽

pp. 3517-3523

Author(s):

CH Hoyt ◽

CJ Oh ◽

JB Beekman ◽

DW Litchfield ◽

KM Lerea

Keyword(s):

Protein Kinase ◽

Protein Kinases ◽

Casein Kinase Ii ◽

Human Platelets ◽

Casein Kinase ◽

Immunogold Electron Microscopy ◽

Beta Subunits ◽

Specific Substrate ◽

Substrate Peptide ◽

Independent Protein

We have recently shown that inhibition of protein phosphatases in platelets causes increases in protein phosphorylations with a concomitant inhibition of platelet responses. The burst in protein phosphorylation appears to be catalyzed by messenger-independent protein kinases. The aim of the present study was to characterize the presence of broad families of protein kinases found in platelets. Lysates of control and thrombin-stimulated platelets were prepared, and proteins were separated on MONO Q fast protein liquid chromatography. In addition to the presence of histone protein kinase and tyrosine kinase activities, human platelets contain casein kinase II (CKII) activity as assessed by phosphorylation of a specific substrate peptide. Western blot analysis and immunogold electron microscopy studies further showed the presence of alpha-, alpha'-, and beta- subunits of CKII. The enzyme appears to be distributed throughout the cytosol and not secreted after thrombin treatment. Immunoprecipitation studies suggest that at least some of the holoenzymes exist as an alpha alpha' beta 2 complex. Although no activation of the enzyme was detected after thrombin treatment, our results show that CKII is a major messenger-independent protein kinase in platelets.

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Coexpression of both .alpha. and .beta. subunits is required for assembly of regulated casein kinase II

Biochemistry ◽

10.1021/bi00110a016 ◽

1991 ◽

Vol 30 (46) ◽

pp. 11133-11140 ◽

Cited By ~ 59

Author(s):

O. Filhol ◽

C. Cochet ◽

P. Wedegaertner ◽

G. N. Gill ◽

E. M. Chambaz

Keyword(s):

Casein Kinase Ii ◽

Casein Kinase ◽

Beta Subunits

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The Schizosaccharomyces pombe casein kinase II alpha and beta subunits: evolutionary conservation and positive role of the beta subunit

Molecular and Cellular Biology ◽

10.1128/mcb.14.1.576-586.1994 ◽

1994 ◽

Vol 14 (1) ◽

pp. 576-586

Author(s):

I Roussou ◽

G Draetta

Keyword(s):

Enzyme Activity ◽

Amino Acid ◽

Amino Acid Sequence ◽

Schizosaccharomyces Pombe ◽

Sequence Similarity ◽

Casein Kinase Ii ◽

Casein Kinase ◽

Beta Subunit ◽

Beta Subunits ◽

High Sequence Similarity

Casein kinase II is a key regulatory enzyme involved in many cellular processes, including the control of growth and cell division. We report the molecular cloning and sequencing of cDNAs encoding the alpha and the beta subunits of casein kinase II of Schizosaccharomyces pombe. The deduced amino acid sequence of Cka1, the alpha catalytic subunit, shows high sequence similarity to alpha subunits identified in other species. The amino acid sequence of Ckb1, the S. pombe beta subunit, is 57% identical to that of the human beta subunit. Cka1 overexpression results in no detectable phenotype. In contrast, Ckb1 overexpression inhibits cell growth and cytokinesis, with formation of multiseptated cells. Disruption of the ckb1+ gene causes a cold-sensitive phenotype and abnormalities in cell shape. In these cells, the casein kinase II activity is reduced to undetectable levels, demonstrating that Ckb1 is required for enzyme activity in vivo. In agreement with this, the activity measured in a strain expressing high levels of Cka1 is enhanced only when the Ckb1 protein is coexpressed. Altogether, our data suggest that Ckb1 is a positive regulator of the enzyme activity, and that it plays a role in mediating the interaction of casein kinase II with downstream targets and/or with additional regulators.

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