Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts

P. R. Wachsberger, W. H. Li, M. Guo, D. Chen, N. Cheong, C. C. Ling, G. Li, G. Iliakis

Research output: Contribution to journalArticle

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Abstract

The role of Ku80 in the repair of DNA double-strand breaks (DSBs) was examined in fibroblasts derived from a Ku80 knockout mouse model described by Nussenzweig et al. (Nature 382, 551-555, 1996). Primary fibroblasts from Ku80(+/+) and Ku80(-/-) mice were immortalized by transfection with plasmids containing either the human MYC proto-oncogene or the Simian virus 40 (SV40) T antigen and were used to measure induction and rejoining of DSBs after exposure to ionizing radiation. The number of DSBs in the cells was quantified by either asymmetric field-inversion gel electrophoresis (AFIGE) or clamped homogeneous electrical-field gel electrophoresis (CHEF). The latter method was introduced for a more reliable quantification of repair even when DNA degradation occurs in a fraction of the irradiated cell population during the postirradiation incubation time. The results confirm that Ku80-deficient mouse fibroblasts are sensitive to ionizing radiation and demonstrate that the increased radiosensitivity may result from a deficiency in DSB rejoining. The results further indicate that unless techniques are employed that allow for distinction between DNA degradation and DNA repair, erroneous conclusions may be drawn regarding the potential of cells to repair DSBs.

Original languageEnglish (US)
Pages (from-to)398-407
Number of pages10
JournalRadiation Research
Volume151
Issue number4
DOIs
StatePublished - 1999

Fingerprint

Double-Stranded DNA Breaks
fibroblasts
strands
mice
deoxyribonucleic acid
Fibroblasts
DNA repair
Ionizing Radiation
ionizing radiation
gel electrophoresis
DNA
Simian virus 40
proto-oncogenes
electrophoresis
degradation
Proto-Oncogenes
Pulsed Field Gel Electrophoresis
Radiation Tolerance
Viral Tumor Antigens
electric field

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Radiology Nuclear Medicine and imaging
  • Biophysics
  • Radiation

Cite this

Wachsberger, P. R., Li, W. H., Guo, M., Chen, D., Cheong, N., Ling, C. C., ... Iliakis, G. (1999). Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts. Radiation Research, 151(4), 398-407. https://doi.org/10.2307/3579826

Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts. / Wachsberger, P. R.; Li, W. H.; Guo, M.; Chen, D.; Cheong, N.; Ling, C. C.; Li, G.; Iliakis, G.

In: Radiation Research, Vol. 151, No. 4, 1999, p. 398-407.

Research output: Contribution to journalArticle

Wachsberger, PR, Li, WH, Guo, M, Chen, D, Cheong, N, Ling, CC, Li, G & Iliakis, G 1999, 'Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts', Radiation Research, vol. 151, no. 4, pp. 398-407. https://doi.org/10.2307/3579826
Wachsberger PR, Li WH, Guo M, Chen D, Cheong N, Ling CC et al. Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts. Radiation Research. 1999;151(4):398-407. https://doi.org/10.2307/3579826
Wachsberger, P. R. ; Li, W. H. ; Guo, M. ; Chen, D. ; Cheong, N. ; Ling, C. C. ; Li, G. ; Iliakis, G. / Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts. In: Radiation Research. 1999 ; Vol. 151, No. 4. pp. 398-407.
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