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.
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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.
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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.
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