Evolution of the Creatine Kinases (original) (raw)
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The primary structure of chicken B-creatine kinase and evidence for heterogeneity of its mRNA
Nucleic Acids Research, 1986
cDNA clones for chicken B-CK were isolated by immunoscreening from a gizzard cDNA library constructed in the expression vector \gtll. The entire coding portion in addition to the complete 3' untranslated region and 42 bp of the 5' noncoding part are represented in the clone H$. On RNA blots H4 insert DNA hybridized to a 1600 bp poly(A) RNA from gizzard, brain and heart but not to breast or skeletal muscle RNA. In vitro generated sense strand transcripts of H4 insert DNA were translated in vitro into a protein indistinguishable from isolated, authentic B-CK. The distinct nucleotide sequences of H4 insert DNA and M-CK cDNA were translated into 82 % homologous amino acid sequences. Sequence heterogeneity among the B-CK cDNA clones within both the 3' noncoding and even in the coding region indicates the existence of multiple B-CK mRNA species.
Molecular cloning and the complete nucleotide sequence of the creatine kinase-M cDNA from chicken
Nucleic Acids Research, 1984
Tn¥ complete nucleotide sequence of mRNA for B-subunit of rat brain S-100 protein was determined from recombinant cDNA clones. The sequence was composed of 1488 bp which Included the 276 bp of the complete coding region, the 120 bp of the 5'-noncoding region and the 1092 bp of the 3'-noncoding region containing two polyadenylation signals. In addition, the poly(A) tail was also found. The amino add sequence deduced from the nucleotide sequence was homologous to the amino acid sequence of bovine S-100 B subunit except 4 residues showing species differences. From the viewpoint of evolutionary implications, the homology between the nucleotide sequence of S-100 and those of rat intestinal Ca-b1nding protein (ICaBP) and calmodulin (CaM) was examined. A dot-blot hybridization of poly(A) RNA from the developing rat brains using a labeled cDNA showed a rapid increase in S-100 mRNA at 10-20 postnatal days. The presence of S-100 mRNA in C-6 glioma cells 1s also described. © IRL Prett Limited, Oxford, England. 7455 by guest on June 18, 2016 http://nar.oxfordjournals.org/ Downloaded from
Molecular cloning of a DNA sequence complementary to creatine kinase M mRNA from chickens
Proceedings of the National Academy of Sciences, 1982
We have cloned and identified a DNA sequence complementary to the mRNA of creatine kinase (CK) isozyme M, although the mRNA is a minor species of the total mRNA in developing myoblasts. Poly(A)+RNA from breast and thigh muscle of 5-week-old chicks was enriched for CK mRNA by a novel procedure of sucrose gradient centrifugation in the presence of methylmercuric hydroxide. DNA complementary to this mRNA was inserted into pBR322, and colonies containing the recombinant plasmids were screened for the ability of the plasmid DNA to hybridize with and rescue CK mRNA from total muscle mRNA. Three plasmids, pCS195, pCS192, and pM35-4, could specifically rescue CK-M mRNA. CK-M mRNA was detected by in vitro translation and specific immunoprecipitation. The identity of the in vitro translation product was further confirmed by its migration in two-dimensional gels at the isoelectric point and molecular weight ofCK-M. The heterogeneity ofCK-M observed in vivo also was found upon translation of the CK-M mRNA which hybridizes to the plasmid.
Molecular characterization of the creatine kinases and some historical perspectives
Bioenergetics of the Cell: Quantitative Aspects, 1998
Over the last 15 years, molecular characterization of the creatine kinase (CK) gene family has paralleled the molecular revolution of understanding gene structure, function, and regulation. In this review, we present a summary of advances in molecular analysis of the CK gene family with a few vignettes of historical interest. We describe how the muscle CK gene provided an essential model system to examine myogenic regulatory mechanisms, leading to the discovery of the binding site for the MyoD family of basic helix-loop-helix transcription factors essential in skeletal myogenesis and the characterization of the MEF2 family of factors with an A/T rich consensus binding site essential in skeletal myogenesis and cardiogenesis. Cloning and characterization of the four mRNAs and nuclear genes encoding the cytosolic CKs, muscle and brain CKs, and the mitochondrial (Mt) CKs, sarcomeric MtCK and ubiquitous MtCK, has allowed intriguing study of tissue-specific and cell-specific expression of the different CKs and analysis of structural, functional, regulatory, and evolutionary relationships among both the four CK proteins and genes. Current and future studies focus on understanding both cellular energetics facilitated by the CK enzymes, especially energy channelling from the site of production, the mitochondrial matrix and inner membrane, to various cytosolic foci of utilization, and regulation of MtCK gene expression at the cell and tissue-specific level as models of regulation of energy producing genes. (Mol Cell Biochem 184: [153][154][155][156][157][158][159][160][161][162][163][164][165][166][167] 1998)
Creatine Kinase Protein Sequence Encoded by a cDNA Made from Torpedo californica Electric Organ mRNA
Proceedings of The National Academy of Sciences, 1984
Creatine kinase (ATP creatine N-phosphotransferase, EC 2.7.3.2) is important in the maintenance of ATP levels in high energy-requiring tissues such as muscle and brain. A complete understanding of its function requires knowledge of its amino acid sequence. To obtain cDNA clones encoding creatine kinase sequences, a cDNA bank was constructed using mRNA from the electric organ of Torpedo californica and was screened by comparing differential colony hybridization of electric organ and liver-derived 32P-labeled cDNAs. Cloned DNAs have been isolated that can arrest the abundant synthesis of Mr 40,000-43,000 material seen after in vitro translation of electric organ mRNA. One of the clones, CK52g8, was sequenced by the dideoxy M13 method and was found to encode a Mr 42,941 protein, which is 68% homologous to a known partial sequence of rabbit muscle creatine kinase and which has a composition similar to creatine kinases from chicken and rabbit tissues. By contrast, no significant homology was found with the known sequences of kinases that use other substrates. RNA blot hybridization analysis indicated that CK52g8 is complementary to a 1600-base-pair mRNA. Primer extension analysis indicated that CK52g8 is only 5 nucleotides short of a full-length cDNA, implying that it encodes a complete protein sequence. The availability of this complete sequence should be useful in further studies of creatine kinase structure and function using techniques such as site-specific mutagenesis.
Gene, 2001
In vertebrates, the creatine kinase (CK) family consists of two cytosolic and two mitochondrial isoforms. The two cytosolic isoforms are the muscle type (M-CK) and the brain type (B-CK). Here we report multiple CK isoenzymes in the diploid channel catfish (Ictalurus punctatus) with one unusual cathodic isoform that was previously found only in pathological situations in human. The cathodic CK isoform existed only in the channel catfish stomach, ovary, and spleen, but not in any other species analyzed such as tilapia, smallmouth bass, chicken, or rat. Two genes encode the multiple forms of the channel catfish M-CK cDNAs. M-CK1 has three alleles, M-CK1.1, M-CK1.2, and M-CK1.3, while M-CK2 has just one allele as determined by analysis of 17 cDNA clones and by allele-specific PCR. M-CK1 encodes a protein of 381 amino acids and the M-CK2 cDNA encodes a protein of 380 amino acids. The two cDNAs shared an 86% identity and both have the nine diagnostic boxes for cytosolic CKs and thus are of cytosolic origin. The M-CK1 gene was isolated, sequenced, and characterized and its promoter should be useful for transgenic research for muscle-specific expression.
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1995
We report that several different chicken and rabbit creatine kinase (CK) J isoenzymes showed an incorporation of 32p when incubated with [7-32p]ATP in an autophosphorylation assay. This modification was shown to be of covalent nature and resulted from an intramolecular phosphorylation reaction that was not dependent on the CK enzymatic activity. By limited proteolysis and sequence analysis of the resulting peptides, the autophosphorylation sites of chicken brain-type CK could be localized within the primary sequence of the enzyme to a 4.5 kDa peptide, spanning a region that is very likely an essential part of the active site of creatine kinase. Homologous peptides were found to be autophosphorylated in chicken muscle-type CK and a mitochondrial CK isoform. Phosphopeptide as well as mutant enzyme analysis provided evidence that threonine-282 2, threonine-289 and serine-285 are involved in the autophosphorylation of CK. Thr-282 and Ser-285 are located close to the reactive cysteine-283. Thr-289 is located within a conserved glycine-rich region highly homologous to the glycine-rich loop of protein kinases, which is known to be important for ATP binding. Thus, it seems likely that the described region constitutes an essential part of the active site of CK.