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DNA Damage-Inducible and RAD52-Independent Repair of DNA Double-Strand Breaks in Saccharomyces cerevisiae
Carol Wood Moorea, Judith McKoya, Michelle Dardalhonb, Darline Davermanna, Marcia Martineza, and Dietrich Averbeckba Department of Microbiology and Immunology, City University of New York Medical School/Sophie Davis School of Biomedical Education and Graduate Programs in Biochemistry and Biology, New York, New York 10031
b Institut Curie-Section de Recherche, CNRS UMR2027, Centre Universitaire d'Orsay, Cedex, F-91405 Orsay, France
Corresponding author: Carol Wood Moore, Department of Microbiology and Immunology, City University of New York Medical School/SDSBE, Science Bldg., Rm. 919, Convent Ave. at 138th St., New York, NY 10031., moore{at}med.cuny.edu (E-mail)
Communicating editor: L. S. SYMINGTON
irradiation or radiomimetic bleomycin, except after high bleomycin doses when chromosomes from rad52/rad52 strains contained fewer DSBs than chromosomes from RAD52/RAD52 strains. DNAs from both genotypes exhibited quick rejoining following irradiation and sedimentation in isokinetic alkaline sucrose gradients, but only chromosomes from RAD52/RAD52 strains exhibited slower rejoining (10 min to 4 hr in growth medium). Chromosomal DSBs introduced by irradiation and bleomycin were analyzed after pulsed-field gel electrophoresis. After equitoxic damage by both DNA-damaging agents, chromosomes in rad52/rad52 cells were reconstructed under nongrowth conditions [liquid holding (LH)]. Up to 100% of DSBs were eliminated and survival increased in RAD52/RAD52 and rad52/rad52 strains. After low doses, chromosomes were sometimes degraded and reconstructed during LH. Chromosomal reconstruction in rad52/rad52 strains was dose dependent after irradiation, but greater after high, rather than low, bleomycin doses with or without LH. These results suggest that a threshold of DSBs is the requisite signal for DNA-damage-inducible repair, and that nonhomologous end-joining repair or another repair function is a dominant mechanism in S. cerevisiae when homologous recombination is impaired.
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