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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
24
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pubmed:dateCreated |
1996-8-20
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pubmed:abstractText |
Three proteins known to play a critical role in mammalian DNA double-strand break repair and lymphoid V(D)J recombination are the autoantigens Ku86 and Ku70 and a 465-kDa serine/threonine protein kinase catalytic subunit (DNA-PKcs). These proteins physically associate to form a complex (DNA.PK) with DNA-dependent protein kinase activity. In this study, we demonstrate using electrophoretic mobility shift assays (EMSAs) that the nuclear DNA end-binding activity of Ku is altered in the human promyelocytic leukemic HL-60 cell line. Western blot and EMSA supershift analyses revealed that HL-60 cells expressed both full-length and variant Ku86 proteins. However, a combined EMSA and immunoanalysis revealed that the Ku heterodimers complexed with DNA in HL-60 cells contained only the variant Ku86 proteins. Finally, UV cross-linking experiments and DNA.PK assays demonstrated that the Ku complexes containing variant Ku86 had a greatly reduced ability to interact with DNA-PKcs and that consequently HL-60 cells had severely diminished DNA.K activity. These data provide important insights into the interaction between Ku and DNA-PKcs and into the role of DNA.PK in DNA double-strand break repair.
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pubmed:grant |
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Nuclear,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Helicases,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Probes,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Activated Protein Kinase,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Ku autoantigen,
http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/PRKDC protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Protein-Serine-Threonine Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors,
http://linkedlifedata.com/resource/pubmed/chemical/Tumor Suppressor Protein p53,
http://linkedlifedata.com/resource/pubmed/chemical/XRCC5 protein, human
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
14
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pubmed:volume |
271
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
14098-104
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pubmed:dateRevised |
2011-4-12
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pubmed:meshHeading |
pubmed-meshheading:8662896-Amino Acid Sequence,
pubmed-meshheading:8662896-Animals,
pubmed-meshheading:8662896-Antibodies, Monoclonal,
pubmed-meshheading:8662896-Antigens, Nuclear,
pubmed-meshheading:8662896-Base Sequence,
pubmed-meshheading:8662896-Blotting, Western,
pubmed-meshheading:8662896-Cell Nucleus,
pubmed-meshheading:8662896-DNA, Neoplasm,
pubmed-meshheading:8662896-DNA Helicases,
pubmed-meshheading:8662896-DNA Probes,
pubmed-meshheading:8662896-DNA-Activated Protein Kinase,
pubmed-meshheading:8662896-DNA-Binding Proteins,
pubmed-meshheading:8662896-Genetic Variation,
pubmed-meshheading:8662896-HL-60 Cells,
pubmed-meshheading:8662896-Humans,
pubmed-meshheading:8662896-Immunoblotting,
pubmed-meshheading:8662896-Mammals,
pubmed-meshheading:8662896-Molecular Sequence Data,
pubmed-meshheading:8662896-Nuclear Proteins,
pubmed-meshheading:8662896-Peptide Fragments,
pubmed-meshheading:8662896-Protein Binding,
pubmed-meshheading:8662896-Protein-Serine-Threonine Kinases,
pubmed-meshheading:8662896-Transcription Factors,
pubmed-meshheading:8662896-Tumor Suppressor Protein p53,
pubmed-meshheading:8662896-Ultraviolet Rays
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pubmed:year |
1996
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pubmed:articleTitle |
Characterization of a Ku86 variant protein that results in altered DNA binding and diminished DNA-dependent protein kinase activity.
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pubmed:affiliation |
Department of Molecular Biology, Brown University, Providence, Rhode Island 02912, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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