title: Remarkably Long-Tract Gene Conversion Induced by Fragile Site Instability in Saccharomyces cerevisiae.
abstract: Replication stress causes breaks at chromosomal locations called common fragile sites. Deletions causing loss of heterozygosity (LOH) in human tumors are strongly correlated with common fragile sites, but the role of gene conversion in LOH at fragile sites in tumors is less well studied. Here, we investigated gene conversion stimulated by instability at fragile site FS2 in the yeast Saccharomyces cerevisiae In our screening system, mitotic LOH events near FS2 are identified by production of red/white sectored colonies. We analyzed single nucleotide polymorphisms between homologs to determine the cause and extent of LOH. Instability at FS2 increases gene conversion 48- to 62-fold, and conversions unassociated with crossover represent 6-7% of LOH events. Gene conversion can result from repair of mismatches in heteroduplex DNA during synthesis-dependent strand annealing (SDSA), double-strand break repair (DSBR), and from break-induced replication (BIR) that switches templates [double BIR (dBIR)]. It has been proposed that SDSA and DSBR typically result in shorter gene-conversion tracts than dBIR. In cells under replication stress, we found that bidirectional tracts at FS2 have a median length of 40.8 kb and a wide distribution of lengths; most of these tracts are not crossover-associated. Tracts that begin at the fragile site FS2 and extend only distally are significantly shorter. The high abundance and long length of noncrossover, bidirectional gene-conversion tracts suggests that dBIR is a prominent mechanism for repair of lesions at FS2, thus this mechanism is likely to be a driver of common fragile site-stimulated LOH in human tumors.
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197.
authors.forename: Shahana A
authors.initials: SA
authors.lastname: Chumki
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197.
authors.forename: Mikael K
authors.initials: MK
authors.lastname: Dunn
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Coates
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197.
authors.forename: Jeanmarie D
authors.initials: JD
authors.lastname: Mishler
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197.
authors.forename: Ellen M
authors.initials: EM
authors.lastname: Younkin
authors.affiliation: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan 48197 anne.casper@emich.edu.
authors.forename: Anne M
authors.identifier: http://orcid.org/0000-0003-4975-5749
authors.initials: AM
authors.lastname: Casper
mesh_terms: D019457:Chromosome Breakage; D043283:Chromosome Fragile Sites; D015825:Chromosomes, Fungal; D003434:Crossing Over, Genetic; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004261:DNA Replication; D004271:DNA, Fungal / Q000235:genetics; D005785:Gene Conversion / Q000235:genetics; D019656:Loss of Heterozygosity; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins
keywords: BIR; fragile site; gene conversion; homologous recombination; loss of heterozygosity