Ku selectively transfers between DNA molecules with homologous ends.
Double strand break repair and V(D)J recombination in mammalian cells require the function of the Ku protein complex and the DNA-dependent protein kinase. The DNA-dependent protein kinase is targeted to DNA through its interaction with the Ku protein complex, and thus the specificity of template recognition in the repair and recombination reactions depend on Ku. We have studied Ku binding to DNA using competitive gel shift analysis. We find that Ku bound to one DNA molecule can transfer directly to another DNA molecule when the two DNA molecules have homologous ends containing a minimum of four matched bases. This remarkable reaction can give a false impression of sequence specificity of Ku DNA binding under certain assay conditions. A model is proposed for the DNA binding function of Ku on the basis of these results and the discovery of a novel type of DNA-Ku complex formed at high Ku/DNA ratios is discussed.
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Cancer Research Campaign Cell Transformation Research Group, Department of Biochemistry, University of Dundee, Dundee DD1 4HN, United Kingdom.
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D034961:Antigens, Nuclear; D001324:Autoantigens / Q000378:metabolism*; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism*; D004265:DNA Helicases; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D000072200:Ku Autoantigen; D008956:Models, Chemical; D009687:Nuclear Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D014157:Transcription Factors / Q000378:metabolism*
D034961:Antigens, Nuclear; D001324:Autoantigens; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D004247:DNA; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen