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 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.

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

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.

scholarly journals Amino acid sequence of the beta subunit of bovine lung casein kinase II.

1987 ◽  
Vol 84 (14) ◽  
pp. 4851-4855 ◽  
Author(s):  
K. Takio ◽  
E. A. Kuenzel ◽  
K. A. Walsh ◽  
E. G. Krebs
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Beta Subunit

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

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.

Identification of the gene encoding 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid desulfhydrase in Burkholderia sp. HME13

2020 ◽  
Vol 85 (3) ◽  
pp. 626-629
Author(s):  
Hisashi Muramatsu ◽  
Hiroki Maguchi ◽  
Taisuke Harada ◽  
Takehiro Kashiwagi ◽  
Chul-Sa Kim ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Propionic Acid ◽  
Unknown Function ◽  
Protein Family ◽  
Amino Acid Sequence Analysis ◽  
Gene Encoding

ABSTRACT Here, we report the identification of the gene encoding a novel enzyme, 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid desulfhydrase, in Burkholderia sp. HME13. The enzyme converts 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid and H2O to 3-(2,5-dioxoimidazolidin-4-yl) propionic acid and H2S. Amino acid sequence analysis of the enzyme indicates that it belongs to the DUF917 protein family, which consists of proteins of unknown function.

scholarly journals Riboflavin synthases of Bacillus subtilis. Purification and amino acid sequence of the alpha subunit.

1990 ◽  
Vol 265 (8) ◽  
pp. 4204-4209
Author(s):  
K Schott ◽  
J Kellermann ◽  
F Lottspeich ◽  
A Bacher
Keyword(s):  
Bacillus Subtilis ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Alpha Subunit
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