Molecular characterization of a human DNA kinase

Karimi-Busheri, F., Daly, G., Robins, P., Canas, B., Pappin, D. J., Sgouros, J., Miller, G. G., Fakhrai, H., Davis, E. M., Le Beau, M. M., Weinfeld, M. (August 1999) Molecular characterization of a human DNA kinase. Journal of Biological Chemistry, 274 (34). pp. 24187-94. ISSN 0021-9258 (Print)0021-9258 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/10446193
DOI: 10.1074/jbc.274.34.24187

Abstract

Human polydeoxyribonucleotide kinase is an enzyme that has the capacity to phosphorylate DNA at 5'-hydroxyl termini and dephosphorylate 3'-phosphate termini and, therefore, can be considered a putative DNA repair enzyme. The enzyme was purified from HeLa cells. Amino acid sequence was obtained for several tryptic fragments by mass spectrometry. The sequences were matched through the dbEST data base with an incomplete human cDNA clone, which was used as a probe to retrieve the 5'-end of the cDNA sequence from a separate cDNA library. The complete cDNA, which codes for a 521-amino acid protein (57.1 kDa), was expressed in Escherichia coli, and the recombinant protein was shown to possess the kinase and phosphatase activities. Comparison with other sequenced proteins identified a P-loop motif, indicative of an ATP-binding domain, and a second motif associated with several different phosphatases. There is reasonable sequence similarity to putative open reading frames in the genomes of Caenorhabditis elegans and Schizosaccharomyces pombe, but similarity to bacteriophage T4 polynucleotide kinase is limited to the kinase and phosphatase domains noted above. Northern hybridization revealed a major transcript of approximately 2.3 kilobases and a minor transcript of approximately 7 kilobases. Pancreas, heart, and kidney appear to have higher levels of mRNA than brain, lung, or liver. Confocal microscopy of human A549 cells indicated that the kinase resides predominantly in the nucleus. The gene encoding the enzyme was mapped to chromosome band 19q13.4.

Item Type: Paper
Additional Information: Karimi-Busheri, F Daly, G Robins, P Canas, B Pappin, D J Sgouros, J Miller, G G Fakhrai, H Davis, E M Le Beau, M M Weinfeld, M CA40046/CA/NCI NIH HHS/United States Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states The Journal of biological chemistry J Biol Chem. 1999 Aug 20;274(34):24187-94.
Uncontrolled Keywords: Amino Acid Sequence Animals Base Sequence Chromosome Mapping DNA, Complementary/isolation & purification HeLa Cells Humans Molecular Sequence Data Polynucleotide 5'-Hydroxyl-Kinase/chemistry/ genetics/isolation & purification Rabbits
Subjects: bioinformatics > genomics and proteomics > design > amino acid design
organism description > animal > C elegans
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > cDNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mRNA
CSHL Authors:
Communities: CSHL labs > Pappin lab
Depositing User: Kathleen Darby
Date: 20 August 1999
Date Deposited: 28 Apr 2014 14:49
Last Modified: 28 Apr 2014 14:49
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29834

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