The repair of double-strand breaks and S1 nuclease-sensitive sites can be monitored chromosome-specifically in Saccharomyces cerevisiae using pulse-field gel electrophoresis.
Repair under non-growth conditions of DNA double-stranded breaks (DSBs) and S1 nuclease-sensitive sites (SSSs; e.g. DNA damage which is processed by in vitro treatment with S1 nuclease to DSBs) induced by [60Co]-gamma-rays (200 Gy; anoxic conditions) was monitored in a diploid repair-competent strain of Saccharomyces cerevisiae. We used pulsed-field gel electrophoresis (PFGE), which allows the separation of chromosome-sized yeast DNA molecules, to determine the number of DSBs and SSSs in individual chromosome species of yeast. Our results indicate that SSSs which have been regarded as clusters of base damage in opposite DNA strands are repaired efficiently in a repair-proficient diploid strain of yeast. The time course of SSS repair is comparable to the one of DSB repair, indicating similarities in the molecular mechanism. Both types of repair kinetics are different for different chromosome species.
Geigl
E M
EM
Gesellschaft für Strahlen- und Umweltforschung mbH, Institut für Strahlenbiologie, Neuherberg, Germany.
Eckardt-Schupp
F
F
D000693:Anaerobiosis / Q000528:radiation effects; D015825:Chromosomes, Fungal; D004249:DNA Damage; D004260:DNA Repair; D004271:DNA, Fungal / Q000096:biosynthesis; D004586:Electrophoresis / Q000379:methods; D005720:Gamma Rays; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
D004271:DNA, Fungal; D015719:Single-Strand Specific DNA and RNA Endonucleases