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DNA Strand Break Rejoining Defect in xrs-6 Is Complemented by Transfection with the Human Ku80 Gene¹

Radiotherapy Research Unit, Institute of Cancer Research, 15 Cotswold Road, Sutton, Surrey SM2 5NG [G. M. R., J. J. E., N. P. M., C. B., G. G. J., T. J. M.], and MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, Sussex, [P. A. J.], United Kingdom

The radiosensitive mutant xrs-6, derived from Chinese hamster ovary cell line CHO-K1, has been demonstrated to be defective in DNA double-strand break repair and also in its proficiency to undergo V(D)J recombination. Recent work has provided both genetic and biochemical evidence that the M_r 80,000 subunit of the Ku protein is able to complement the radiosensitivity and the V(D)J recombination defect in the xrs-6 mutant. We demonstrate here that complementation of the radiosensitive phenotype in xrs-6 cells by the introduction of Ku80 cDNA is accompanied by the concomitant restoration of DNA double-strand break rejoining proficiency to almost that of the parental CHO-K1 cells, as measured both by neutral single-cell microgel electrophoresis (Comet) technique and by pulsed-field gel electrophoresis. These results provide further biochemical evidence for the involvement of the Ku protein in the repair of DNA double-strand breaks.

¹ This work was supported by a Medical Research Council Clinical Science Fellowship (to G. M. R.) and the Cancer Research Campaign.

² To whom requests for reprints should be addressed.

Received 12/27/94. Accepted 2/ 2/95.

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