At the start of this project, the PI (Corentin Claeys Bouuaert) set up his laboratory at the Host Institution (UCLouvain, Belgium) and recruited a team of researchers to work on the four aims of the proposal.
(1) To gain insight into meiotic DSB formation, we developed for the first time a biochemical system that recapitulates the DNA cleavage activity of Spo11. We also investigated the DNA-binding properties of the Spo11 core complex and performed structure-function analyses. Finally, we characterized the structure and function of RMM proteins and their role in the assembly of the DSB machinery by DNA-driven condensation.
(2) We investigated the biochemical properties of meiotic axis proteins Hop1 and Red1 and their relationships with the meiotic DSB proteins. In addition, we characterized the interactions between the DSB protein Mer2 and the histone H3K4me3 reader, Spp1.
(3) To investigate the mechanism of crossover formation, we have made progress in developing an in vitro system using purified crossover-promoting proteins and a novel DNA substrate that mimics a key recombination intermediate, the double Holliday Junction.
(4) Finally, we set up new mass-spectrometry based assays to identify proteins associated with meiotic recombination.
Some of the results from the BrokenGenome project have already been published and presented at international conferences. Currently, the team is finalizing a number of publications describing the bulk of the achievement of the project. Together, the BrokenGenome project has brought important new insights into the molecular processes involved in the formation and repair of DNA double-strand breaks during meiosis.
