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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
12
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pubmed:dateCreated |
1991-1-8
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pubmed:abstractText |
A DNA-activated protein kinase (DNA-PK) was purified from nuclei of HeLa cells. Activity was associated with a single high-molecular-mass (approximately-300,000 Da) polypeptide when analyzed by gel filtration, denaturing polyacrylamide gel electrophoresis, and Western immunoblotting using a monoclonal antibody that also inhibits enzyme activity. Nuclear localization was indicated by subcellular fractionation and confirmed by immunofluorescence on whole cells. Double-stranded DNA stimulated phosphorylation of the 300-kDa polypeptide in purified preparations as well as phosphorylation of the exogenous substrates alpha-casein, simian virus 40 large T antigen, and the human heat shock protein hsp90. Autophosphorylation led to inactivation of the enzyme. The phosphorylation of casein was stimulated over 30-fold by DNA and was specific for serine and threonine residues. Bovine serum albumin and histone H1 were poor substrates for DNA-PK, and no phosphorylation of immunoglobulin G or histones other than H1 was observed. Supercoiled or heat-denatured DNA and synthetic double-stranded RNA or RNA-DNA copolymers did not stimulate casein phosphorylation by DNA-PK. Interaction of the enzyme with DNA in the absence of exogenous substrates was demonstrated by thermal inactivation and gel mobility shifts. These characteristics identify DNA-PK as distinct from other protein kinases described in the literature and suggest that activation by DNA is an important feature of the enzyme's in vivo function.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-207704,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-210162,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2188729,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-222266,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2247067,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2475255,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2507541,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2531292,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2576632,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2668302,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2683077,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2752037,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2770543,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-282611,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2844417,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2893795,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2956925,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2964277,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2992362,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-2998755,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3034432,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3052279,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3091258,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3113737,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3202865,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3370672,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3422454,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3456346,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-3949752,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-4018025,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-4084312,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-4144105,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-4336,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-4345911,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6184076,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6244264,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6244279,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6501318,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-652516,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6717433,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-6828386,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-7251604,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-7274455,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2247066-893433
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0270-7306
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
6460-71
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:2247066-Antibodies, Monoclonal,
pubmed-meshheading:2247066-Blotting, Western,
pubmed-meshheading:2247066-Cell Nucleus,
pubmed-meshheading:2247066-Chromatography, Affinity,
pubmed-meshheading:2247066-Chromatography, Gel,
pubmed-meshheading:2247066-Chromatography, Ion Exchange,
pubmed-meshheading:2247066-DNA,
pubmed-meshheading:2247066-Fluorescent Antibody Technique,
pubmed-meshheading:2247066-HeLa Cells,
pubmed-meshheading:2247066-Humans,
pubmed-meshheading:2247066-Immunoglobulin G,
pubmed-meshheading:2247066-Kinetics,
pubmed-meshheading:2247066-Molecular Weight,
pubmed-meshheading:2247066-Polydeoxyribonucleotides,
pubmed-meshheading:2247066-Protein Kinases
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pubmed:year |
1990
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pubmed:articleTitle |
A DNA-activated protein kinase from HeLa cell nuclei.
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pubmed:affiliation |
Department of Biological Sciences, St. John's University, Jamaica, New York 11439.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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