Mouse PRDM9 DNA-Binding Specificity Determines Sites of Histone H3 Lysine 4 Trimethylation for Initiation of Meiotic Recombination

Abstract

The nature of the PRDM9 zinc finger domain determines the location of hotspots for meiotic recombination in the genome and promotes local histone H3K4 trimethylation.

Abstract

Meiotic recombination generates reciprocal exchanges between homologous chromosomes (also called crossovers, COs) that are essential for proper chromosome segregation during meiosis and are a major source of genome diversity by generating new allele combinations. COs have two striking properties: they occur at specific sites, called hotspots, and these sites evolve rapidly. In mammals, the Prdm9 gene, which encodes a meiosis-specific histone H3 methyltransferase, has recently been identified as a determinant of CO hotspots. Here, using transgenic mice, we show that the sole modification of PRDM9 zinc fingers leads to changes in hotspot activity, histone H3 lysine 4 trimethylation (H3K4me3) levels, and chromosome-wide distribution of COs. We further demonstrate by an in vitro assay that the PRDM9 variant associated with hotspot activity binds specifically to DNA sequences located at the center of the three hotspots tested. Remarkably, we show that mutations in cis located at hotspot centers and associated with a decrease of hotspot activity affect PRDM9 binding. Taken together, these results provide the direct demonstration that Prdm9 is a master regulator of hotspot localization through the DNA binding specificity of its zinc finger array and that binding of PRDM9 at hotspots promotes local H3K4me3 enrichment.

Author Summary

Meiosis is the process of cell division that reduces the number of chromosome sets from two to one, so producing gametes for sexual reproduction. During meiosis in many organisms, there is reciprocal exchange of genetic material between homologous chromosomes by the formation of “crossovers,” which promote genetic diversity by creating new combinations of gene variants and play an important mechanical role in the segregation of chromosomes. Crossovers do not occur randomly throughout the genome, but in small regions called hotspots. Recent work showed that hotspots have specific structural features and that the protein PRDM9 is important in specifying their location. PRDM9 contains a so-called zinc finger domain that is predicted to bind specific DNA sequences, suggesting that hotspots might be sites where PRDM9 binds. By using transgenic mice expressing PRDM9 with modified zinc fingers, here we show directly that the nature of the zinc fingers in PRDM9 determines crossover hotspot localization. We show that PRDM9 binds DNA sequences at the center of hotspots. Furthermore, we identify DNA sequence polymorphisms that affect its binding and the extent of crossover activity. Overall, our work shows that PRDM9, through its zinc finger domain, is a master regulator of hotspot location in the mouse genome.