Journal Article

ATM and PRDM9 regulate SPO11-bound recombination intermediates during meiosis

Maman, Yaakov, Yamada, Shintaro, Wu, Wei, Paola Cotrim, Ana, Paull, Tanya T., Sciascia, Nicholas, Day, Amanda, Paiano, Jacob, Deshpande, Rajashree A., Callen, Elsa, Nussenzweig, André

2020

Overview

Meiotic recombination is initiated by SPO11-induced double-strand breaks (DSBs). In most mammals, the methyltransferase PRDM9 guides SPO11 targeting, and the ATM kinase controls meiotic DSB numbers. Following MRE11 nuclease removal of SPO11, the DSB is resected and loaded with DMC1 filaments for homolog invasion. Here, we demonstrate the direct detection of meiotic DSBs and resection using END-seq on mouse spermatocytes with low sample input. We find that DMC1 limits both minimum and maximum resection lengths, whereas 53BP1, BRCA1 and EXO1 play surprisingly minimal roles. Through enzymatic modifications to END-seq, we identify a SPO11-bound meiotic recombination intermediate (SPO11-RI) present at all hotspots. We propose that SPO11-RI forms because chromatin-bound PRDM9 asymmetrically blocks MRE11 from releasing SPO11. In Atm –/– spermatocytes, trapped SPO11 cleavage complexes accumulate due to defective MRE11 initiation of resection. Thus, in addition to governing SPO11 breakage, ATM and PRDM9 are critical local regulators of mammalian SPO11 processing. Recombination requires DNA break formation by SPO11, following which SPO11 is thought to be released. Here, the authors show that meiotic hotspots retain SPO11 through a recombination intermediate dependent on the methyltransferase PRDM9, and that the ATM kinase governs the release of SPO11.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 49/23 / 631/337/1427 / 631/337/149 / Animals / Ataxia Telangiectasia Mutated Proteins - genetics / Ataxia Telangiectasia Mutated Proteins - metabolism / BRCA1 protein / BRCA1 Protein - genetics / BRCA1 Protein - metabolism / Cell Cycle Proteins - genetics / Cell Cycle Proteins - metabolism / Chromatin / DNA damage / DNA Repair Enzymes - genetics / DNA Repair Enzymes - metabolism / Endodeoxyribonucleases - genetics / Endodeoxyribonucleases - metabolism / Exodeoxyribonucleases - genetics / Exodeoxyribonucleases - metabolism / Female / Filaments / Gametocytes / Histone-Lysine N-Methyltransferase - genetics / Histone-Lysine N-Methyltransferase - metabolism / Homologous Recombination - physiology / Homology / Humanities and Social Sciences / Intermediates / Kinases / Male / Mammals / Meiosis / Meiosis - physiology / Methyltransferase / Mice / Mice, Inbred C57BL / Mice, Knockout / MRE11 Homologue Protein - metabolism / Mre11 nuclease / MRE11 protein / multidisciplinary / Nuclease / Phosphate-Binding Proteins - genetics / Phosphate-Binding Proteins - metabolism / Recombination / Regulators / Science / Science (multidisciplinary) / Spermatocytes / Spermatocytes - metabolism / Tumor Suppressor p53-Binding Protein 1 - genetics / Yeast