Participation of DNA-PKcs in DSB repair after exposure to high- and low-LET radiation.
Cellular lesions (e.g. DSBs) are induced into DNA upon exposure to radiation, with DSB complexity increasing with radiation ionization density. Using M059K and M059J human glioblastoma cells (proficient and deficient in DNA-PKcs activity, respectively), we investigated the repair of DNA damage, including DSBs, induced by high- and low-LET radiation [gamma rays, alpha particles and high-charge and energy (HZE) ions]. In the absence of DNA-PKcs activity, less DSB repair and increased recruitment of RAD51 was seen at 24 h. After exposure to (56)Fe heavy ions, the number of cells with RAD51 tracks was less than the number of cells with gamma-H2AX at 24 h with both cell lines. Using alpha particles, comparable numbers of cells with visible gamma-H2AX and RAD51 were seen at 24 h in both cell lines. M059J cells irradiated with alpha particles accumulated in S phase, with a greater number of cyclin A and RAD51 co-stained cells seen at 24 h compared with M059K cells, where an S-phase block is absent. It is proposed that DNA-PKcs plays a role in the repair of some frank DSBs, which are longer-lived in NHEJ-deficient cells, and some non-DSB clustered damage sites that are converted into DSBs at replication as the cell cycles through to S phase.
Anderson
Jennifer A
JA
DNA Damage Group, Gray Institute for Radiation Oncology and Biology, University of Oxford, Oxford, OX3 7DQ, United Kingdom.
Harper
Jane V
JV
Cucinotta
Francis A
FA
O'Neill
Peter
P
D045744:Cell Line, Tumor; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair / Q000528:radiation effects; D051747:DNA-Activated Protein Kinase / Q000302:isolation & purification / Q000378:metabolism / Q000528:radiation effects*; D016521:Electrophoresis, Gel, Pulsed-Field; D005347:Fibroblasts / Q000201:enzymology / Q000502:physiology / Q000528:radiation effects; D005720:Gamma Rays / Q000009:adverse effects*; D005909:Glioblastoma; D006801:Humans; D007504:Iron Radioisotopes / Q000494:pharmacology*; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism / Q000528:radiation effects; D011829:Radiation Dosage
D007504:Iron Radioisotopes; D051747:DNA-Activated Protein Kinase; D011493:Protein Kinase C