Double-strand breaks (DSBs) are the most deleterious lesions experienced by our genome. Yet, DSBs are intentionally induced during gamete formation to promote the exchange of genetic material between homologous chromosomes. While the conserved topoisomerase-like enzyme Spo11 catalyzes DSBs, additional regulatory proteins-referred to as "Spo11 accessory factors"- regulate the number, timing, and placement of DSBs during early meiotic prophase ensuring that SPO11 does not wreak havoc on the genome. Despite the importance of the accessory factors, they are poorly conserved at the sequence level suggesting that these factors may adopt unique functions in different species. In this work, we present a detailed analysis of the genetic and physical interactions between the DSB factors in the nematode Caenorhabditis elegans providing new insights into conserved and novel functions of these proteins. This work shows that HIM-5 is the determinant of X-chromosome-specific crossovers and that its retention in the nucleus is dependent on DSB-1, the sole accessory factor that interacts with SPO-11. We further provide evidence that HIM-5 coordinates the actions of the different accessory factors sub-groups, providing insights into how components on the DNA loops may interact with the chromosome axis.

Department of Biochemistry and Biophysics University of Pennsylvania Penn Center for Genome Integrity Philadelphia Pennsylvania

Department of Biology Masaryk University Czech Republic

Department of Computational and Systems Biology University of Pittsburgh School of Medicine Pittsburgh PA 15213 USA

Department of Genetics Blavatnik Institute Harvard Medical School 77 Avenue Louis Pasteur Room 334 Boston MA 02115 USA

Department of Microbiology and Molecular Genetics University of Pittsburgh School of Medicine UPMC Hillman Cancer Center Pittsburgh Pennsylvania

Department of Obstetrics Gynecology and Reproductive Sciences University of Pittsburgh School of Medicine Pittsburgh PA 15213 USA

Magee Womens Research Institute Pittsburgh PA 15213 USA

Tsinghua U Medical School China

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