DNA strand break rejoining defect in xrs-6 is complemented by transfection with the human Ku80 gene.
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.
Ross
G M
GM
Radiotherapy Research Unit, Institute of Cancer Research, Sutton, Surrey, United Kingdom.
Eady
J J
JJ
Mithal
N P
NP
Bush
C
C
Steel
G G
GG
Jeggo
P A
PA
McMillan
T J
TJ
D000818:Animals; D034961:Antigens, Nuclear; D016466:CHO Cells; D006224:Cricetinae; D004249:DNA Damage; D004265:DNA Helicases; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology; D000072200:Ku Autoantigen; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology; D011836:Radiation Tolerance; D014162:Transfection
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen