Evolutionary conservation of the structure and function of meiotic Rec114-Mei4 and Mer2 complexes

In this study, Daccache et al. describe structural and functional properties of the meiotic DNA double-strand break complex, comprised of Rec114-Mei4 and Mer2 proteins, in S. cerevisiae. They further show that the architectures of Rec114-Mei4 and Mer2, and their multivalent binding to branched DNA, are evolutionarily conserved and important for complex condensation. Meiosis-specific Rec114-Mei4 and Mer2 complexes are thought to enable Spo11-mediated DNA double-strand break (DSB) formation through a mechanism that involves DNA-dependent condensation. However, the structure, molecular properties, and evolutionary conservation of Rec114-Mei4 and Mer2 are unclear. Here, we present AlphaFold models of Rec114-Mei4 and Mer2 complexes supported by nuclear magnetic resonance (NMR) spectroscopy, small-angle X-ray scattering (SAXS), and mutagenesis. We show that dimers composed of the Rec114 C terminus form & alpha;-helical chains that cup an N-terminal Mei4 & alpha; helix, and that Mer2 forms a parallel homotetrameric coiled coil. Both Rec114-Mei4 and Mer2 bind preferentially to branched DNA substrates, indicative of multivalent protein-DNA interactions. Indeed, the Rec114-Mei4 interaction domain contains two DNA-binding sites that point in opposite directions and drive condensation. The Mer2 coiled-coil domain bridges coaligned DNA duplexes, likely through extensive electrostatic interactions along the length of the coiled coil. Finally, we show that the structures of Rec114-Mei4 and Mer2 are conserved across eukaryotes, while DNA-binding properties vary significantly. This work provides insights into the mechanism whereby Rec114-Mei4 and Mer2 complexes promote the assembly of the meiotic DSB machinery and suggests a model in which Mer2 condensation is the essential driver of assembly, with the DNA-binding activity of Rec114-Mei4 playing a supportive role.

Automated summary

In this study, the authors describe the structural and functional properties of the meiotic DNA double-strand break complex. They show that the Rec114-Mei4 and Mer2 proteins are important for complex condensation and that their architectures are evolutionarily conserved. The study presents models of the Rec114-Mei4 and Mer2 complexes supported by various techniques, and reveals their DNA-binding properties.