C. elegans Germ Cells Switch between Distinct Modes of Double-Strand Break Repair During Meiotic Prophase Progression

Abstract

Chromosome inheritance during sexual reproduction relies on deliberate induction of double-strand DNA breaks (DSBs) and repair of a subset of these breaks as interhomolog crossovers (COs). Here we provide a direct demonstration, based on our analysis of rad-50 mutants, that the meiotic program in Caenorhabditis elegans involves both acquisition and loss of a specialized mode of double-strand break repair (DSBR). In premeiotic germ cells, RAD-50 is not required to load strand-exchange protein RAD-51 at sites of spontaneous or ionizing radiation (IR)-induced DSBs. A specialized meiotic DSBR mode is engaged at the onset of meiotic prophase, coincident with assembly of meiotic chromosome axis structures. This meiotic DSBR mode is characterized both by dependence on RAD-50 for rapid accumulation of RAD-51 at DSB sites and by competence for converting DSBs into interhomolog COs. At the mid-pachytene to late pachytene transition, germ cells undergo an abrupt release from the meiotic DSBR mode, characterized by reversion to RAD-50-independent loading of RAD-51 and loss of competence to convert DSBs into interhomolog COs. This transition in DSBR mode is dependent on MAP kinase-triggered prophase progression and coincides temporally with a major remodeling of chromosome architecture. We propose that at least two developmentally programmed switches in DSBR mode, likely conferred by changes in chromosome architecture, operate in the C. elegans germ line to allow formation of meiotic crossovers without jeopardizing genomic integrity. Our data further suggest that meiotic cohesin component REC-8 may play a role in limiting the activity of SPO-11 in generating meiotic DSBs and that RAD-50 may function in counteracting this inhibition. Faithful inheritance of chromosomes during sexual reproduction depends on the deliberate formation of double-strand DNA breaks (DSBs) and subsequent repair of a subset of these breaks by a mechanism that leads to crossovers between homologous chromosome pairs. The requirement for crossovers to ensure chromosome segregation poses a challenge for sexually reproducing organisms, as DSBs constitute a danger to genomic integrity in other contexts. This manuscript provides insight into the mechanisms that allow germ cells to generate recombination-based linkages that ensure chromosome inheritance while at the same time protecting the integrity of their genomes. Specifically, we provide a direct demonstration, based on our analysis of rad-50 mutants, that the meiotic program in C. elegans involves both acquisition and loss of a specialized meiotic mode of double-strand break repair (DSBR). We propose that the ability to revert to a less constrained DSBR environment at a late stage of meiotic prophase serves as a fail-safe mechanism for safeguarding the genome, as it provides an opportunity to repair any remaining DBSs and restore chromosome integrity prior to chromosome segregation.