Molecular cloning of casein kinase II alpha subunit from Dictyostelium discoideum and its expression in the life cycle

1992 ◽  
Vol 12 (12) ◽  
pp. 5711-5723
Author(s):  
U Kikkawa ◽  
S K Mann ◽  
R A Firtel ◽  
T Hunter
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Cdna Clone ◽  
Blot Analysis ◽  
Vegetative Growth ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Developmental Expression ◽  
Alpha Subunit ◽  
Alpha Gene

A Dictyostelium discoideum cDNA encoding an alpha-type subunit of casein kinase II was isolated, and its cDNA was used to study developmental expression of casein kinase II during the Dictyostelium life cycle. The 1.3-kb cDNA insert contained an open reading frame of 337 amino acids (M(r) 39,900). The deduced amino acid sequence has high homology with those of casein kinase II alpha subunits from other species. Genomic Southern blot analysis suggested that there is a single gene encoding casein kinase II alpha subunit in D. discoideum. Northern (RNA) blot analysis showed that the casein kinase II alpha-subunit gene is expressed constitutively as a 1.9-kb mRNA throughout vegetative growth and multicellular development. Casein kinase purified from normal vegetative cells contained a major protein band of approximately 36 kDa, which was recognized by antisera raised against rat testis casein kinase II. Comparison of the in vitro transcription/translation product of the alpha-subunit cDNA clone and the purified 36-kDa protein by partial proteolysis indicated that the isolated cDNA clone encodes the Dictyostelium casein kinase II alpha subunit. No protein corresponding to a beta subunit was detected in purified casein kinase. Immunoblot analysis using anti-rat casein kinase II sera showed that the alpha subunit of casein kinase II is expressed constitutively like its mRNA during the life cycle of D. discoideum. Casein kinase II activity measured by using a specific peptide substrate paralleled the level of alpha subunit detected by immunoblotting during the life cycle, with a maximum variation of approximately 2-fold. We were unable to obtain disruptants of the casein kinase II alpha gene, suggesting that there is a single casein kinase II alpha gene, which is essential for vegetative growth of D. discoideum.

scholarly journals Molecular cloning of casein kinase II alpha subunit from Dictyostelium discoideum and its expression in the life cycle.

1992 ◽  
Vol 12 (12) ◽  
pp. 5711-5723 ◽  
Author(s):  
U Kikkawa ◽  
S K Mann ◽  
R A Firtel ◽  
T Hunter
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Cdna Clone ◽  
Blot Analysis ◽  
Vegetative Growth ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Developmental Expression ◽  
Alpha Subunit ◽  
Alpha Gene

A Dictyostelium discoideum cDNA encoding an alpha-type subunit of casein kinase II was isolated, and its cDNA was used to study developmental expression of casein kinase II during the Dictyostelium life cycle. The 1.3-kb cDNA insert contained an open reading frame of 337 amino acids (M(r) 39,900). The deduced amino acid sequence has high homology with those of casein kinase II alpha subunits from other species. Genomic Southern blot analysis suggested that there is a single gene encoding casein kinase II alpha subunit in D. discoideum. Northern (RNA) blot analysis showed that the casein kinase II alpha-subunit gene is expressed constitutively as a 1.9-kb mRNA throughout vegetative growth and multicellular development. Casein kinase purified from normal vegetative cells contained a major protein band of approximately 36 kDa, which was recognized by antisera raised against rat testis casein kinase II. Comparison of the in vitro transcription/translation product of the alpha-subunit cDNA clone and the purified 36-kDa protein by partial proteolysis indicated that the isolated cDNA clone encodes the Dictyostelium casein kinase II alpha subunit. No protein corresponding to a beta subunit was detected in purified casein kinase. Immunoblot analysis using anti-rat casein kinase II sera showed that the alpha subunit of casein kinase II is expressed constitutively like its mRNA during the life cycle of D. discoideum. Casein kinase II activity measured by using a specific peptide substrate paralleled the level of alpha subunit detected by immunoblotting during the life cycle, with a maximum variation of approximately 2-fold. We were unable to obtain disruptants of the casein kinase II alpha gene, suggesting that there is a single casein kinase II alpha gene, which is essential for vegetative growth of D. discoideum.

Isolation, sequencing, and disruption of the CKA1 gene encoding the alpha subunit of yeast casein kinase II

1988 ◽  
Vol 8 (11) ◽  
pp. 4981-4990
Author(s):  
J L Chen-Wu ◽  
R Padmanabha ◽  
C V Glover
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gene Product ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Protein Sequencing ◽  
Alpha Subunit ◽  
Yeast Genome ◽  
Gene Encoding ◽  
Alpha Gene

Casein kinase II of Saccharomyces cerevisiae contains two distinct catalytic subunits, alpha and alpha', which must be encoded by separate genes (R. Padmanabha and C. V. C. Glover, J. Biol. Chem. 262:1829-1835, 1987). The gene encoding the 42-kilodalton alpha subunit has been isolated by screening a yeast genomic library with oligonucleotide probes synthesized on the basis of the N-terminal amino acid sequence of the polypeptide. This gene (designated CKA1) contains an intron-free open reading frame of 372 amino acid residues. The deduced amino acid sequence is 67% identical to the alpha subunit of Drosophila melanogaster casein kinase II. The CKA1 gene product appears to be distantly related to other known protein kinases but exhibits highest similarity to the CDC28 gene product and its homolog in other species. Gene replacement techniques have been used to generate a null cka1 mutant allele. Haploid and diploid strains lacking a functional CKA1 gene appear to be phenotypically wild type, presumably because of the presence of the alpha' gene. Interestingly, the CKA1 gene appears to be single copy in the yeast genome; i.e., the alpha' gene, whose existence is known from biochemical studies and protein sequencing, cannot be detected by low-stringency hybridization.

scholarly journals Isolation, sequencing, and disruption of the CKA1 gene encoding the alpha subunit of yeast casein kinase II.

1988 ◽  
Vol 8 (11) ◽  
pp. 4981-4990 ◽  
Author(s):  
J L Chen-Wu ◽  
R Padmanabha ◽  
C V Glover
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gene Product ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Protein Sequencing ◽  
Alpha Subunit ◽  
Yeast Genome ◽  
Gene Encoding ◽  
Alpha Gene

Casein kinase II of Saccharomyces cerevisiae contains two distinct catalytic subunits, alpha and alpha', which must be encoded by separate genes (R. Padmanabha and C. V. C. Glover, J. Biol. Chem. 262:1829-1835, 1987). The gene encoding the 42-kilodalton alpha subunit has been isolated by screening a yeast genomic library with oligonucleotide probes synthesized on the basis of the N-terminal amino acid sequence of the polypeptide. This gene (designated CKA1) contains an intron-free open reading frame of 372 amino acid residues. The deduced amino acid sequence is 67% identical to the alpha subunit of Drosophila melanogaster casein kinase II. The CKA1 gene product appears to be distantly related to other known protein kinases but exhibits highest similarity to the CDC28 gene product and its homolog in other species. Gene replacement techniques have been used to generate a null cka1 mutant allele. Haploid and diploid strains lacking a functional CKA1 gene appear to be phenotypically wild type, presumably because of the presence of the alpha' gene. Interestingly, the CKA1 gene appears to be single copy in the yeast genome; i.e., the alpha' gene, whose existence is known from biochemical studies and protein sequencing, cannot be detected by low-stringency hybridization.

Molecular cloning of the human casein kinase II .alpha. subunit [Erratum to document cited in CA110(21):187044x]

Biochemistry ◽  
1989 ◽  
Vol 28 (17) ◽  
pp. 7138-7138
Author(s):  
Herman Meisner ◽  
Robin Heller-Harrison ◽  
Joanne Buxton ◽  
Michael P. Czech
Keyword(s):  
Molecular Cloning ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Alpha Subunit

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

1990 ◽  
Vol 10 (8) ◽  
pp. 4089-4099 ◽  
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.

scholarly journals Immunocytochemical localization of casein kinase II during interphase and mitosis.

1991 ◽  
Vol 114 (6) ◽  
pp. 1217-1232 ◽  
Author(s):  
I J Yu ◽  
D L Spector ◽  
Y S Bae ◽  
D R Marshak
Keyword(s):  
Hela Cells ◽  
Casein Kinase Ii ◽  
Microscopic Investigation ◽  
Electron Microscopic Investigation ◽  
Casein Kinase ◽  
Alpha Subunit ◽  
Electron Microscopic ◽  
Peptide Antigens ◽  
Hydroxyurea Treatment ◽  
The Individual

We have developed specific antibodies to synthetic peptide antigens that react with the individual subunits of casein kinase II (CKII). Using these antibodies, we studied the localization of CKII in asynchronous HeLa cells by immunofluorescence and immunoelectron microscopy. Further studies were done on HeLa cells arrested at the G1/S transition by hydroxyurea treatment. Our results indicate that the CKII alpha and beta subunits are localized in the cytoplasm during interphase and are distributed throughout the cell during mitosis. Further electron microscopic investigation revealed that CKII alpha subunit is associated with spindle fibers during metaphase and anaphase. In contrast, the CKII alpha' subunit is localized in the nucleus during G1 and in the cytoplasm during S. Taken together, our results suggest that CKII may play significant roles in cell division control by shifting its localization between the cytoplasm and nucleus.

Antibodies to casein kinase II in sera of patients with mixed connective tissue disease: evaluation with recombinant proteins

1993 ◽  
Vol 39 (12) ◽  
pp. 2492-2494 ◽  
Author(s):  
C Wiemann ◽  
L Bodenbach ◽  
W Pyerin
Keyword(s):  
Differential Diagnosis ◽  
Connective Tissue ◽  
Connective Tissue Disease ◽  
Recombinant Proteins ◽  
Mixed Connective Tissue Disease ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Alpha Subunit

Abstract In this study we determined the prevalence of autoantibodies against casein kinase II (CKII) in patients positive for anti-70K marker antibodies, which is indicative of mixed connective tissue disease. An anti-CKII ELISA was established with the use of bacterially expressed recombinant CKII proteins. Eight out of 52 anti-70K-positive sera (15%) were positive for anti-CKII antibodies, which recognized preferentially the CKII alpha subunit. All control sera (n = 52) were anti-CKII negative. Thus, the occurrence of anti-CKII antibodies may be of value for differential diagnosis.

Molecular cloning of the human casein kinase II .alpha. subunit

Biochemistry ◽  
1989 ◽  
Vol 28 (9) ◽  
pp. 4072-4076 ◽  
Author(s):  
Herman Meisner ◽  
Robin Heller-Harrison ◽  
Joanne Buxton ◽  
Michael P. Czech
Keyword(s):  
Molecular Cloning ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Alpha Subunit
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