SPO11 dimers are sufficient to catalyse DNA double-strand breaks in vitro.

ABSTRACT: SPO11 initiates meiotic recombination through the induction of programmed DNA double-strand breaks (DSBs)^1,2, but this catalytic activity has never been reconstituted in vitro^3,4. Here, using Mus musculus SPO11, we report a biochemical system that recapitulates all the hallmarks of meiotic DSB formation. We show that SPO11 catalyses break formation in the absence of any partners and remains covalently attached to the 5' broken strands. We find that target site selection by SPO11 is influenced by the sequence, bendability and topology of the DNA substrate, and provide evidence that SPO11 can reseal single-strand DNA breaks. In addition, we show that SPO11 is monomeric in solution and that cleavage requires dimerization for the reconstitution of two hybrid active sites.

SUBMITTER: Oger C 

PROVIDER: S-EPMC11922746 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature