Global Analysis of the Meiotic Crossover Landscape
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ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
PROVIDER: GSE12170 | GEO | 2008/09/16
SECONDARY ACCESSION(S): PRJNA113459
REPOSITORIES: GEO
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Project description:Meiotic crossover formation requires the stabilization of early recombination intermediates by a set of proteins and occurs within the environment of the chromosome axis, a structure important for the regulation of meiotic recombination events. The molecular mechanisms underlying and connecting crossover recombination and axis localization are elusive. Here, we identified the ZZS (Zip2–Zip4–Spo16) complex, required for crossover formation, which carries two distinct activities: one provided by Zip4, which acts as hub through physical interactions with components of the chromosome axis and the crossover machinery, and the other carried by Zip2 and Spo16, which preferentially bind branched DNA molecules in vitro. We found that Zip2 and Spo16 share structural similarities to the structure-specific XPF–ERCC1 nuclease, although it lacks endonuclease activity. The XPF domain of Zip2 is required for crossover formation, suggesting that, together with Spo16, it has a noncatalytic DNA recognition function. Our results suggest that the ZZS complex shepherds recombination intermediates toward crossovers as a dynamic structural module that connects recombination events to the chromosome axis. The identification of the ZZS complex improves our understanding of the various activities required for crossover implementation and is likely applicable to other organisms, including mammals.
2018-02-14 | GSE103877 | GEO
Project description:To study recombination at the fine-scale, we used high-throughput sequencing of 300 to 1,000 crossovers within the RAC1 R gene hotspot. This revealed focused intragenic crossovers, overlapping exons encoding the TIR, NBS and LRR domains. To examine the role of recombination pathways, we repeated this experiment in recq4a recq4b, fancm and recq4a recq4b fancm mutants. Finally, in order to investigate how varying patterns of interhomolog divergence influence local patterns of crossover frequency, we repeated RAC1 pollen typing sequencing in different F1 hybrids.
2018-07-01 | E-MTAB-6333 | biostudies-arrayexpress
Project description:Meiotic crossover recombination is triggered by the formation of programmed double strand breaks (DSBs) catalyzed by the conserved Spo11 protein. Only a subset of these DSBs are repaired as crossovers, promoted by a group of eight evolutionarily conserved proteins, named ZMM. The synaptonemal complex (SC) that assembles between homologous chromosomes is composed of two axial elements, from which chromatin loops emanate, held together by a central region, composed of the central element and a transverse filament protein. In budding yeast, crossover formation is functionally linked to the formation of the SC, but the underlying mechanism is unknown. Here we found that the SC central element protein, Ecm11, mainly localizes on both DSB sites and sites that attach chromatin loops to the chromosome axis, in a way that strictly requires the ZMM protein Zip4. We further show that Zip4 directly interacts with Ecm11 and that point mutants that specifically abolish the Zip4-Ecm11 interaction loose Ecm11 binding to chromosomes and exhibit defective SC assembly. Interestingly, SC assembly can be partially rescued by artificially tethering interaction-defective Ecm11 to Zip4. This direct connection that ensures SC assembly from CO sites could be a way for the meiotic cell to shut down further DSB formation once enough recombination sites have been selected for crossovers, thereby preventing excess crossovers. Finally, we found that the mammalian ortholog of Zip4, TEX11, interacts with the SC central element TEX12, raising the possibility that this may be a general mechanism.
2021-09-01 | GSE177033 | GEO
Project description:Using yeast strains based on the protease-deficient strain BJ5459 (= wild type), the effects of the spt10-null and spt10-C388S mutations on global gene expression were assessed and compared. BJ5459 (wild type) and BJ-SPT10(C388S)-HA cells were grown in synthetic complete medium to A600 = 0.6. RNA was prepared using an RNeasy kit (Qiagen). Biotinylated cRNA was prepared from three independently prepared sample pairs and hybridised to Affymetrix S98 arrays and detected using fluorescently labelled streptavidin. The wild type and the spt10-C388S average signals each represent the average of the signals from three arrays. The P-value is an indicator of the statistical significance of the difference between the average signals for wild type and the C388S mutant. The lower the value, the greater the significance. The gene descriptor is provided by Affymetrix. Keywords: mutant yeast strain comparison
2005-12-22 | GSE3354 | GEO
Project description:To map meiotic crossovers and noncrossovers across the yeast genome, we genotyped the four spores of 51 tetrads arising from sporulation of a YJM789/S96 hybrid strain. Sporulation was induced, tetrads were dissected, and genomic DNA from each spore was hybridized to microarrays, one array per spore. 12 YJM789 and 13 S96 hybridizations were also performed as a reference for genotyping. To analyze recombination mutants, we also hybridized DNA from 5 tetrads arising from a YJM789/S96 hybrid strain homozygous for a MSH4 deletion, and from 20 spores (6 dyads and 8 single spores) arising from a hybrid homozygous for a MMS4 deletion.
2009-08-04 | E-TABM-470 | biostudies-arrayexpress
Project description:Interference exists ubiquitously in many biological processes. Crossover interference patterns meiotic crossovers, which are required for faithful chromosome segregation and evolutionary adaption. However, what the interference signal is and how it is generated and regulated are unknown. We show that yeast top2 alleles cannot bind or cleave DNA accumulate a higher level of negative supercoils and show weaker interference. However, top2 alleles cannot religate the cleaved DNA or release the religated DNA accumulate less negative supercoils and show stronger interference. Moreover, the level of negative supercoils is negatively correlated with crossover interference strength in various strains. Furthermore, negative supercoils preferentially enrich at crossover-associated Zip3 regions before the formation of meiotic DNA double-strand breaks, and regions with more negative supercoils tend to have more Zip3. Additionally, the strength of crossover interference and homeostasis change coordinately in mutants. These findings suggest that the accumulation and relief of negative supercoils pattern meiotic crossovers.
2022-08-24 | GSE201366 | GEO
Project description:Two types of RNA:DNA associations can lead to genome instability: the formation of R-loops during transcription and the incorporation of ribonucleotide monophosphates (rNMPs) into DNA during replication. Both ribonuclease (RNase) H1 and RNase H2 degrade the RNA component of R-loops, whereas only RNase H2 can remove one or a few rNMPs from DNA. We performed high-resolution mapping of mitotic recombination events throughout the yeast genome in diploid strains of Saccharomyces cerevisiae lacking RNase H1 (rnh1Δ), RNase H2 (rnh201Δ), or both RNase H1 and RNase H2 (rnh1Δ rnh201Δ). We found little effect on recombination in the rnh1Δ strain, but elevated recombination in both the rnh201Δ and the double-mutant strains; levels of recombination in the double mutant were about 50% higher than in the rnh201 single-mutant strain. An rnh201Δ mutant that additionally contained a mutation that reduces rNMP incorporation by DNA polymerase ε (pol2-M644L) had a level of instability similar to that observed in the presence of wild-type Polε. This result suggests that the elevated recombination observed in the absence of only RNase H2 is primarily a consequence of R loops rather than misincorporated rNMPs. The details of these experiments are in press in Genetics This diploid S.cerevisiae strain used here is a hybrid between W303-1A and YJM789. The backgrounds are diverged by about 50,000 polymorphisms that are surveyed by custom CGH microarrays containing SNP-specific probes (St. Charles et al., 2012; Genetics 190: 1267-1284; PMID 22267500). The whole genome array surveyed loss of heterozygosity (LOH) events throughout the genome of in wild type, pol2-M644L, rnh1Δ, rnh201Δ, rnh201Δ pol2-M644L, and rnh1Δ rnh201Δ mutant backgrounds in 10, 12, 10, 19, 17, and 22 samples, respectively. The names of these strains are KO_198,KO_234, KO_73, KO_75, KO_244, and KO_5, respectively. Another assay was performed mapping reciprocal crossover events on the right arm of chromosome IV in the rnh201Δ (strains KO_135 and KO188) and the rnh1Δrnh201Δ (strain KO_132) mutant backgrounds. A reporter system with a wild type ADE2 on the end of chromosome IV allowed us to detect crossovers by seeing a red-white sectored colony. Each reciprocal event was mapped by doing a microarray specific for the right arm of chromosome IV (St. Charles and Petes, 2013; PLoS Genet. 1267-1284 PMCID PMC3616911) on the red and white sides of sectors. 21 sectors were analyzed for the rnh201Δ mutant while 14 were analyzed for the rnh1Δ rnh201Δ mutant. We mapped crossovers on the right arm of chromosome IV in the rnh201Δ mutant in four different diploid isolates: KO135_5 (1R/1W, 2R/2W, 4R/4W, 5R/5W, 12R/12W, and 13R/13W), KO_135_6 (1R/1W, 4R/4W, 5R/5W, 7R/7W, 9R/9W, 11R/11W, 13R/13W, and 17R/17W), KO 188_1 (1R/1W, and 4R/4W), and KO188_2 (6R/6W, 7R/7W, 11R/11W, and 12R/12W). We mapped crossovers on the right arm of chromosome IV in the rnh1Δ rnh201Δ mutant in two different diploid isolates: KO_132_31 (2R/2W, 6R/6W, 8R/8W, 15R/15W, 17R/17W, 21R/21W, 22R/22W, and 24R/24W) and KO_132_29 (3R/3W, 7R/7W, 18R/18W, 24R/24W, and 34R/34W).
2015-09-23 | E-GEOD-73334 | biostudies-arrayexpress
Project description:Furfural is a key inhibitor in S. cerevisiae fermentation causing serious economic loss. To disclose of the recombinational DNA lesions induced by furfural, we analyzed 19 JSC25-derived sectored colonies using chromosome IV SNP-specific microarrays. The parent diploid S. cerevisiae strain JSC25 (constructed by crossing two haploid strains isogenic W303-1A and YJM789) was homologous for ade2-1 alleles. One copy of gene SUP4-o, an ochre-suppressing tRNA, was inserted at the end of right arm of chromosome IV. Under normal conditions, the colony color of JSC25 is pink. A crossover event occurred on the right arm of chromosome IV at the first cell cycle would result in a white/red sectored colony on solid plate.
2017-10-13 | GSE104880 | GEO
Project description:Crossover recombination is a hallmark of meiosis, which holds the paternal and maternal chromosomes (homologs) together for their faithful separation, meanwhile, it promotes genetic diversity of progenies. The pattern of crossover is mainly controlled by the architecture of meiotic chromosomes. Environmental factors, especially temperature, also play an important role in modulating crossovers. However, it is unclear how temperature affects crossovers. Here, we examined the distributions of budding yeast axis components (Red1, Hop1, and Rec8) and the CO-associated Zip3 foci in detail in different temperatures, and found that both increased and decreased temperatures result in shorter meiotic chromosome axes and more crossovers. Further investigations showed that altered temperature coordinately enhanced the hyperabundant accumulation of Hop1 and Red1 on chromosomes and the number of Zip3 foci. Most importantly, temperature-induced alterations in axis distribution and Zip3 foci depend on the changes in DNA negative supercoil. These findings suggest that yeast meiosis senses temperature changes by increasing the level of negative supercoil to increase crossovers and modulate chromosome organization. These findings provide a novel view in understanding the effect and mechanism of temperature on meiosis recombination and chromosome organization, and thus also have an important implication in evolution and breeding.
2024-07-30 | GSE246901 | GEO
Project description:During meiosis, crossover recombination is essential to link homologous chromosomes and drive faithful chromosome segregation. Crossover recombination is non-random across the genome, and centromere-proximal crossovers are associated with an increased risk of aneuploidy, including Trisomy 21 in humans. Here, we identify the conserved Ctf19/CCAN kinetochore sub-complex as a major factor that minimizes potentially deleterious centromere-proximal crossovers in budding yeast. We uncover multi-layered suppression of pericentromeric recombination by the Ctf19 complex, operating across distinct chromosomal distances. The Ctf19 complex prevents meiotic DNA break formation, the initiating event of recombination, proximal to the centromere. The Ctf19 complex independently drives the enrichment of cohesin throughout the broader pericentromere to suppress crossovers, but not DNA breaks. This non-canonical role of the kinetochore in defining a chromosome domain that is refractory to crossovers adds a new layer of functionality by which the kinetochore prevents the incidence of chromosome segregation errors that generate aneuploid gametes. Two samples total: two biological replicate Spo11-oligo maps of S. cerevisiae SK1 mcm21 null mutant
2015-12-15 | E-GEOD-72683 | biostudies-arrayexpress