Double-strand break repair in the absence of RAD51 in yeast: a possible role for break-induced DNA replication

• Published in: Proceedings of the National Academy of Sciences - PNAS 1996-07, Vol.93 (14), p.7131-7136
• Main Authors: Malkova, A. (Brandeis University, Waltham, MA.), Ivanov, E.L, Haber, J.E
• Format: Article
• Language: English
• Subjects: ADN; Bacteria; Cells; CHROMOSOME; Chromosomes; Chromosomes, Fungal; CROMOSOMAS; Crossovers; Daughter cells; DIPLOIDIA; DIPLOIDIE; Diploidy; DNA; DNA Damage; DNA Repair; DNA Replication; DNA, Fungal - biosynthesis; DNA, Fungal - genetics; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Fungal Proteins - metabolism; GENE; Gene Conversion; GENES; Genetic loci; Genetics; Homozygote; LOCI; LOCUS; Mating Factor; MUTANT; MUTANTES; Peptides - genetics; PROTEINAS AGLUTINANTES; PROTEINE DE LIAISON; Rad51 Recombinase; RECOMBINACION; RECOMBINAISON; Recombination, Genetic; REPLICACION; REPLICATION; SACCHAROMYCES CEREVISIAE; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; Telomere; Templates, Genetic; Yeast; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.14.7131

In wild-type diploid cells of Saccharomyces cerevisiae, an HO endonuclease-induced double-strand break (DSB) at the MAT locus can be efficiently repaired by gene conversion using the homologous chromosome sequences. Repair of the broken chromosome was nearly eliminated in rad52 delta diploids; 99% lost the broken chromosome. However, in rad51 delta diploids, the broken chromosomes were repaired approximately 35% of the time. None of these repair events were simple gene conversions or gene conversions with an associated crossover; instead, they created diploids homozygous for the MAT locus and all markers in the 100-kb region distal to the site of the DSB. In rad51 delta diploids, the broken chromosome can apparently be inherited for several generations, as many of these repair events are found as sectored colonies, with one part being repaired and the other part having lost the broken chromosome. Similar events occur in about 2% of wild-type cells. We propose that a broken chromosome end can invade a homologous template in the absence of RAD51 and initiate DNA replication that may extend to the telomere, 100 or more kb away. Such break-induced replication appears to be similar to recombination-initiated replication in bacteria