Differential radiosensitivity phenotypes of DNA-PKcs mutations affecting NHEJ and HRR systems following irradiation with gamma-rays or very low fluences of alpha particles

Yu Fen Lin, Hatsumi Nagasawa, John B. Little, Takamitsu A. Kato, Hung Ying Shih, Xian Jin Xie, Paul F. Wilson, John R. Brogan, Akihiro Kurimasa, David J. Chen, Joel S. Bedford, Benjamin P C Chen

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Abstract

We have examined cell-cycle dependence of chromosomal aberration induction and cell killing after high or low dose-rate γ irradiation in cells bearing DNA-PKcs mutations in the S2056 cluster, the T2609 cluster, or the kinase domain. We also compared sister chromatid exchanges (SCE) production by very low fluences of α-particles in DNA-PKcs mutant cells, and in homologous recombination repair (HRR) mutant cells including Rad51C, Rad51D, and Fancg/xrcc9. Generally, chromosomal aberrations and cell killing by γ-rays were similarly affected by mutations in DNA-PKcs, and these mutant cells were more sensitive in G1 than in S/G2 phase. In G1-irradiated DNA-PKcs mutant cells, both chromosome- and chromatid-type breaks and exchanges were in excess than wild-type cells. For cells irradiated in late S/G2 phase, mutant cells showed very high yields of chromatid breaks compared to wild-type cells. Few exchanges were seen in DNA-PKcs-null, Ku80-null, or DNA-PKcs kinase dead mutants, but exchanges in excess were detected in the S2506 or T2609 cluster mutants. SCE induction by very low doses of α-particles is resulted from bystander effects in cells not traversed by α-particles. SCE seen in wild-type cells was completely abolished in Rad51C- or Rad51D-deficient cells, but near normal in Fancg/xrcc9 cells. In marked contrast, very high levels of SCEs were observed in DNA-PKcs-null, DNA-PKcs kinase-dead and Ku80-null mutants. SCE induction was also abolished in T2609 cluster mutant cells, but was only slightly reduced in the S2056 cluster mutant cells. Since both non-homologous end-joining (NHEJ) and HRR systems utilize initial DNA lesions as a substrate, these results suggest the possibility of a competitive interference phenomenon operating between NHEJ and at least the Rad51C/D components of HRR; the level of interaction between damaged DNA and a particular DNA-PK component may determine the level of interaction of such DNA with a relevant HRR component.

Original languageEnglish (US)
Article numbere93579
JournalPLoS One
Volume9
Issue number4
DOIs
StatePublished - Apr 8 2014
Externally publishedYes

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Alpha Particles
Recombinational DNA Repair
Alpha particles
Radiation Tolerance
homologous recombination
Gamma Rays
Joining
Gamma rays
gamma radiation
Repair
irradiation
Irradiation
Phenotype
mutation
phenotype
Mutation
DNA
mutants
cells
Sister Chromatid Exchange

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Differential radiosensitivity phenotypes of DNA-PKcs mutations affecting NHEJ and HRR systems following irradiation with gamma-rays or very low fluences of alpha particles. / Lin, Yu Fen; Nagasawa, Hatsumi; Little, John B.; Kato, Takamitsu A.; Shih, Hung Ying; Xie, Xian Jin; Wilson, Paul F.; Brogan, John R.; Kurimasa, Akihiro; Chen, David J.; Bedford, Joel S.; Chen, Benjamin P C.

In: PLoS One, Vol. 9, No. 4, e93579, 08.04.2014.

Research output: Contribution to journalArticle

Lin, YF, Nagasawa, H, Little, JB, Kato, TA, Shih, HY, Xie, XJ, Wilson, PF, Brogan, JR, Kurimasa, A, Chen, DJ, Bedford, JS & Chen, BPC 2014, 'Differential radiosensitivity phenotypes of DNA-PKcs mutations affecting NHEJ and HRR systems following irradiation with gamma-rays or very low fluences of alpha particles', PLoS One, vol. 9, no. 4, e93579. https://doi.org/10.1371/journal.pone.0093579
Lin, Yu Fen ; Nagasawa, Hatsumi ; Little, John B. ; Kato, Takamitsu A. ; Shih, Hung Ying ; Xie, Xian Jin ; Wilson, Paul F. ; Brogan, John R. ; Kurimasa, Akihiro ; Chen, David J. ; Bedford, Joel S. ; Chen, Benjamin P C. / Differential radiosensitivity phenotypes of DNA-PKcs mutations affecting NHEJ and HRR systems following irradiation with gamma-rays or very low fluences of alpha particles. In: PLoS One. 2014 ; Vol. 9, No. 4.
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