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rdf:type |
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lifeskim:mentions |
umls-concept:C0012854,
umls-concept:C0033684,
umls-concept:C0040715,
umls-concept:C0205171,
umls-concept:C0205263,
umls-concept:C0205431,
umls-concept:C0271510,
umls-concept:C0439855,
umls-concept:C0444930,
umls-concept:C0449829,
umls-concept:C0599718,
umls-concept:C0599813,
umls-concept:C0599893,
umls-concept:C1158537,
umls-concept:C1335262,
umls-concept:C1337080,
umls-concept:C1522702
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pubmed:issue |
24
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pubmed:dateCreated |
2000-1-25
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pubmed:abstractText |
Ku protein and the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) are essential components of the double-strand break repair machinery in higher eukaryotic cells. Ku protein binds to broken DNA ends and recruits DNA-PKcs to form an enzymatically active complex. To characterize the arrangement of proteins in this complex, we developed a set of photocross-linking probes, each with a single free end. We have previously used this approach to characterize the contacts in an initial Ku-DNA complex, and we have now applied the same technology to define the events that occur when Ku recruits DNA-PKcs. The new probes allow the binding of one molecule of Ku protein and one molecule of DNA-PKcs in a defined position and orientation. Photocross-linking reveals that DNA-PKcs makes direct contact with the DNA termini, occupying an approximately 10 bp region proximal to the free end. Characterization of the Ku protein cross-linking pattern in the presence and absence of DNA-PKcs suggests that Ku binds to form an initial complex at the DNA ends, and that recruitment of DNA-PKcs induces an inward translocation of this Ku molecule by about one helical turn. The presence of ATP had no effect on protein-DNA contacts, suggesting that neither DNA-PK-mediated phosphorylation nor a putative Ku helicase activity plays a role in modulating protein conformation under the conditions tested.
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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/Antigens, Nuclear,
http://linkedlifedata.com/resource/pubmed/chemical/Cross-Linking Reagents,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
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/Protein-Serine-Threonine Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/XRCC5 protein, human
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
1362-4962
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:day |
15
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pubmed:volume |
27
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
4679-86
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:10572166-Antigens, Nuclear,
pubmed-meshheading:10572166-Base Sequence,
pubmed-meshheading:10572166-Binding Sites,
pubmed-meshheading:10572166-Cell Nucleus,
pubmed-meshheading:10572166-Cross-Linking Reagents,
pubmed-meshheading:10572166-DNA,
pubmed-meshheading:10572166-DNA Damage,
pubmed-meshheading:10572166-DNA Helicases,
pubmed-meshheading:10572166-DNA Probes,
pubmed-meshheading:10572166-DNA Repair,
pubmed-meshheading:10572166-DNA-Activated Protein Kinase,
pubmed-meshheading:10572166-DNA-Binding Proteins,
pubmed-meshheading:10572166-HeLa Cells,
pubmed-meshheading:10572166-Humans,
pubmed-meshheading:10572166-Kinetics,
pubmed-meshheading:10572166-Models, Molecular,
pubmed-meshheading:10572166-Nuclear Proteins,
pubmed-meshheading:10572166-Nucleic Acid Conformation,
pubmed-meshheading:10572166-Protein Structure, Quaternary,
pubmed-meshheading:10572166-Protein-Serine-Threonine Kinases
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pubmed:year |
1999
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pubmed:articleTitle |
Geometry of a complex formed by double strand break repair proteins at a single DNA end: recruitment of DNA-PKcs induces inward translocation of Ku protein.
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pubmed:affiliation |
Program in Gene Regulation, Institute of Molecular Medicine and Genetics, Medical College of Georgia, 1120 15th Street, Augusta, GA 30912, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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