<p>In <i>Drosophila melanogaster</i> females, as in most organisms, the segregation of homologous chromosomes in meiosis depends on the formation of crossovers between them. In most cases, crossovers require the synaptonemal complex (SC), a conserved multi-protein structure that forms between homologous chromosomes in early meiosis. Recent studies leveraging partial-loss-of-function alleles suggest that the SC plays a more direct role in crossover formation. One SC protein that is involved in crossover formation is SYP-4 in nematodes, which we found is a likely ortholog of the <i>D. melanogaster</i> SC protein Corolla. To create a hypomorphic allele of <i>corolla</i> in <i>D. melanogaster</i>, we used CRISPR/Cas9 to replace it with its <i>D. mauritiana</i> ortholog, yielding <i>corolla</i><sup><i>mau</i></sup>. Since SC protein sequences are rapidly diverging while maintaining the SC’s structure, we hypothesized that this replacement would enable SC assembly but show defects in crossover formation. Indeed, at 25&#xa0;°C <i>corolla</i><sup><i>mau</i></sup> homozygous females exhibited defects in SC maintenance and crossover formation, resulting in moderate levels of chromosome missegregation. At 18&#xa0;°C, SC maintenance was rescued, and recombination rates were improved, although they remained significantly lower than observed in wild type. Unexpectedly, in homozygotes we also observed unique polycomplexes composed of the SC proteins Corolla and Corona but lacking the transverse filament protein C(3)G. Overall, we report a novel hypomorphic allele of <i>corolla</i> that suggests Corolla regulates crossover formation. Further, the unique polycomplexes found in mutant flies may provide new insights into SC architecture and protein-protein interactions.</p>

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The synaptonemal complex component corolla regulates meiotic crossover formation in Drosophila melanogaster

  • Stefanie Williams,
  • Grace McKown,
  • Zulin Yu,
  • Jennifer Gardner,
  • Cynthia Staber,
  • Matthew C. Gibson,
  • R. Scott Hawley

摘要

In Drosophila melanogaster females, as in most organisms, the segregation of homologous chromosomes in meiosis depends on the formation of crossovers between them. In most cases, crossovers require the synaptonemal complex (SC), a conserved multi-protein structure that forms between homologous chromosomes in early meiosis. Recent studies leveraging partial-loss-of-function alleles suggest that the SC plays a more direct role in crossover formation. One SC protein that is involved in crossover formation is SYP-4 in nematodes, which we found is a likely ortholog of the D. melanogaster SC protein Corolla. To create a hypomorphic allele of corolla in D. melanogaster, we used CRISPR/Cas9 to replace it with its D. mauritiana ortholog, yielding corollamau. Since SC protein sequences are rapidly diverging while maintaining the SC’s structure, we hypothesized that this replacement would enable SC assembly but show defects in crossover formation. Indeed, at 25 °C corollamau homozygous females exhibited defects in SC maintenance and crossover formation, resulting in moderate levels of chromosome missegregation. At 18 °C, SC maintenance was rescued, and recombination rates were improved, although they remained significantly lower than observed in wild type. Unexpectedly, in homozygotes we also observed unique polycomplexes composed of the SC proteins Corolla and Corona but lacking the transverse filament protein C(3)G. Overall, we report a novel hypomorphic allele of corolla that suggests Corolla regulates crossover formation. Further, the unique polycomplexes found in mutant flies may provide new insights into SC architecture and protein-protein interactions.