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Department of Radiation Medicine, Division of Radiation Research, Georgetown University Medical Center, Washington, DC [D. P., M. J., A. D.]; Department of Physics, The George Washington University, Washington, DC [D. P.]; and Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, Georgia [S. Y., W. S. D.]
The binding of the Ku protein to DNA was investigated using the atomic force microscope. Ku was found to bind predominantly to the ends of double-stranded DNA. Experiments with plasmid DNA revealed that Ku does not bind to circular plasmids but does bind to plasmids that have been linearized by treatment with ionizing radiation. The binding of Ku to poly(dG-dC)-poly(dG-dC) polynucleotides and to a 400-bp DNA EcoRI fragment resulted in a shift in the fragment size distribution to include longer fragments, with internally binding Ku. Furthermore, we observed images consistent with fragments joined together by Ku, showing an interaction with two ends of DNA. These observations suggest that Ku may play a role in physically orienting DNA for ligation by binding the ends of adjacent DNA molecules.
1 This work was supported in part by Grants CA45408 (to A. D.) and GM 35866 (to W. S. D.) from the NIH.
2 To whom requests for reprints should be addressed, at Department of Radiation Medicine, Division of Radiation Research, Georgetown University Medical Center, The Research Building, E216A, 3970 Reservoir Road NW, Washington, DC 20007-2197. Phone: (202) 687-2144; Fax: (202) 687-0400.
Received 12/27/96. Accepted 3/ 8/97.
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