A Yeast Two-Hybrid Screen for SYP-3 Interactors Identifies SYP-4, a Component Required for Synaptonemal Complex Assembly and Chiasma Formation in Caenorhabditis elegans Meiosis

Abstract

The proper assembly of the synaptonemal complex (SC) between homologs is critical to ensure accurate meiotic chromosome segregation. The SC is a meiotic tripartite structure present from yeast to humans, comprised of proteins assembled along the axes of the chromosomes and central region (CR) proteins that bridge the two chromosome axes. Here we identify SYP-4 as a novel structural component of the SC in Caenorhabditis elegans. SYP-4 interacts in a yeast two-hybrid assay with SYP-3, one of components of the CR of the SC, and is localized at the interface between homologs during meiosis. SYP-4 is essential for the localization of SYP-1, SYP-2, and SYP-3 CR proteins onto chromosomes, thereby playing a crucial role in the stabilization of pairing interactions between homologous chromosomes. In the absence of SYP-4, the levels of recombination intermediates, as indicated by RAD-51 foci, are elevated in mid-prophase nuclei, and crossover recombination events are significantly reduced. The lack of chiasmata observed in syp-4 mutants supports the elevated levels of chromosome nondisjunction manifested in high embryonic lethality. Altogether our findings place SYP-4 as a central player in SC formation and broaden our understanding of the structure of the SC and its assembly.

Author Summary

Meiosis is a two-part cell division program that ensures the formation of haploid gametes (e.g. eggs and sperm), which can then reconstitute a species' ploidy through fertilization. A critical step towards accomplishing this task is the accurate segregation of homologous chromosomes away from each other during meiosis I. This requires the formation of at least one obligatory crossover event (genetic exchange) between each pair of homologous chromosomes. In most organisms, the formation of all crossover events greatly relies on the synaptonemal complex (SC). This “zipper-like” structure holds the pairs of homologous chromosomes together during meiotic prophase I, and crossover recombination is completed in the context of the fully formed SCs. Here, we identify SYP-4 as a novel structural component of the SC in the nematode C. elegans. In its absence, SCs fail to form, resulting in a lack of crossover formation and increased errors in chromosome segregation. SYP-4 interacts in a yeast two-hybrid assay with SYP-3, one of the SC proteins, and its localization onto chromosomes is interdependent with SYP-1, SYP-2, and SYP-3 proteins. SYP-4 therefore plays a critical role during C. elegans meiosis in generating the ultrastructurally conserved SC that is ubiquitously present from yeast to humans.