Reconstitution of SPO11-dependent double-strand break formation.

ABSTRACT: Meiotic recombination starts with SPO11 generation of DNA double-strand breaks (DSBs)¹. SPO11 is critical for meiosis in most species, but it generates dangerous DSBs with mutagenic² and gametocidal³ potential. Cells must therefore utilize the beneficial functions of SPO11 while minimizing its risks⁴-how they do so remains poorly understood. Here we report reconstitution of DNA cleavage in vitro with purified recombinant mouse SPO11 bound to TOP6BL. SPO11-TOP6BL complexes are monomeric (1:1) in solution and bind tightly to DNA, but dimeric (2:2) assemblies cleave DNA to form covalent 5' attachments that require SPO11 active-site residues, divalent metal ions and SPO11 dimerization. SPO11 can also reseal DNA that it has nicked. Structure modelling

SUBMITTER: Zheng Z 

PROVIDER: S-EPMC11922745 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature