Recombinogenic conditions influence partner choice in spontaneous mitotic recombination.
Mammalian common fragile sites are loci of frequent chromosome breakage and putative recombination hotspots. Here, we utilized Replication Slow Zones (RSZs), a budding yeast homolog of the mammalian common fragile sites, to examine recombination activities at these loci. We found that rates of URA3 inactivation of a hisG-URA3-hisG reporter at RSZ and non-RSZ loci were comparable under all conditions tested, including those that specifically promote chromosome breakage at RSZs (hydroxyurea [HU], mec1Δ sml1Δ, and high temperature), and those that suppress it (sml1Δ and rrm3Δ). These observations indicate that RSZs are not recombination hotspots and that chromosome fragility and recombination activity can be uncoupled. Results confirmed recombinogenic effects of HU, mec1Δ sml1Δ, and rrm3Δ and identified temperature as a regulator of mitotic recombination. We also found that these conditions altered the nature of recombination outcomes, leading to a significant increase in the frequency of URA3 inactivation via loss of heterozygosity (LOH), the type of genetic alteration involved in cancer development. Further analyses revealed that the increase was likely due to down regulation of intrachromatid and intersister (IC/IS) bias in mitotic recombination, and that RSZs exhibited greater sensitivity to HU dependent loss of IC/IS bias than non RSZ loci. These observations suggest that recombinogenic conditions contribute to genome rearrangements not only by increasing the overall recombination activity, but also by altering the nature of recombination outcomes by their effects on recombination partner choice. Similarly, fragile sites may contribute to cancer more frequently than non-fragile loci due their enhanced sensitivity to certain conditions that down-regulate the IC/IS bias rather than intrinsically higher rates of recombination.
Stem Cell Biology and Developmental Genetics, National Institute for Medical Research, MRC, The Ridgeway, London, United Kingdom.
James D
JD
Cauwood
Anthony L
AL
Johnson
Alexander
A
Widger
Rita S
RS
Cha
D019457:Chromosome Breakage / Q000187:drug effects; D043283:Chromosome Fragile Sites / Q000187:drug effects / Q000235:genetics*; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics*; D006918:Hydroxyurea / Q000494:pharmacology; D008938:Mitosis / Q000235:genetics*; D009154:Mutation; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics
D006918:Hydroxyurea