Radiation induction of delayed recombination in Schizosaccharomyces pombe.
Ionizing radiation is known to induce delayed chromosome and gene mutations in the descendants of the irradiated tissue culture cells. Molecular mechanisms of such delayed mutations are yet to be elucidated, since high genomic complexity of mammalian cells makes it difficult to analyze. We now tested radiation induction of delayed recombination in the fission yeast Schizosaccharomyces pombe by monitoring the frequency of homologous recombination after X-irradiation. A reporter with 200 bp tandem repeats went through spontaneous recombination at a frequency of 1.0 x 10(-4), and the frequency increased dose-dependently to around 10 x 10(-4) at 500 Gy of X-irradiation. Although the repair of initial DNA damage was thought to be completed before the restart of cell division cycle, the elevation of the recombination frequency persisted for 8-10 cell generations after irradiation (delayed recombination). The delayed recombination suggests that descendants of the irradiated cells keep a memory of the initial DNA damage which upregulates recombination machinery for 8-10 generations even in the absence of DNA double-strand breaks (DSBs). Since radical scavengers were ineffective in inhibiting the delayed recombination, a memory by continuous production of DNA damaging agents such as reactive oxygen species (ROS) was excluded. Recombination was induced in trans in a reporter on chromosome III by a DNA DSB at a site on chromosome I, suggesting the untargeted nature of delayed recombination. Interestingly, Rad22 foci persisted in the X-irradiated population in parallel with the elevation of the recombination frequency. These results suggest that the epigenetic damage memory induced by DNA DSB upregulates untargeted and delayed recombination in S. pombe.
Takeda
Jun
J
Department of Radiation System Biology, Radiation Biology Center, Kyoto University, Yoshida Konoe, Kyoto, Japan.
Uematsu
Norio
N
Shiraishi
Satomi
S
Toyoshima
Megumi
M
Matsumoto
Tomohiro
T
Niwa
Ohtsura
O
D001483:Base Sequence; D002453:Cell Cycle; D004249:DNA Damage; D004271:DNA, Fungal; D004268:DNA-Binding Proteins / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D017382:Reactive Oxygen Species / Q000378:metabolism; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism; D014965:X-Rays
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D017382:Reactive Oxygen Species; D029702:Schizosaccharomyces pombe Proteins; C085412:rad52 protein, S pombe