title: Repeat expansion in the budding yeast ribosomal DNA can occur independently of the canonical homologous recombination machinery.
abstract: Major eukaryotic genomic elements, including the ribosomal DNA (rDNA), are composed of repeated sequences with well-defined copy numbers that must be maintained by regulated recombination. Although mechanisms that instigate rDNA recombination have been identified, none are directional and they therefore cannot explain precise repeat number control. Here, we show that yeast lacking histone chaperone Asf1 undergo reproducible rDNA repeat expansions. These expansions do not require the replication fork blocking protein Fob1 and are therefore independent of known rDNA expansion mechanisms. We propose the existence of a regulated rDNA repeat gain pathway that becomes constitutively active in asf1Δ mutants. Cells lacking ASF1 accumulate rDNA repeats with high fidelity in a processive manner across multiple cell divisions. The mechanism of repeat gain is dependent on highly repetitive sequence but, surprisingly, is independent of the homologous recombination proteins Rad52, Rad51 and Rad59. The expansion mechanism is compromised by mutations that decrease the processivity of DNA replication, which leads to progressive loss of rDNA repeats. Our data suggest that a novel mode of break-induced replication occurs in repetitive DNA that is dependent on high homology but does not require the canonical homologous recombination machinery.
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3JR, UK. jon.houseley@bbsrc.ac.uk
authors.forename: Jonathan
authors.initials: J
authors.lastname: Houseley
authors.forename: David
authors.initials: D
authors.lastname: Tollervey
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics; D042622:DNA Repeat Expansion; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis / Q000737:chemistry*; D004275:DNA, Ribosomal / Q000096:biosynthesis / Q000737:chemistry*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D017353:Gene Deletion; D018832:Molecular Chaperones / Q000235:genetics; D009154:Mutation; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: C109731:ASF1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; D004259:DNA-Directed DNA Polymerase