PRDM9 activity depends on HELLS and promotes local 5-hydroxymethylcytosine enrichment
- National Institute of Genetics, Japan
- CNRS UM, France
- CNRS INSERM UM, France
Abstract
Meiotic recombination starts with the formation of DNA double-strand breaks (DSBs) at specific genomic locations that correspond to PRDM9 binding sites. The molecular steps occurring from PRDM9 binding to DSB formation are unknown. Using proteomic approaches to find PRDM9 partners, we identified HELLS, a member of the SNF2-like family of chromatin remodelers. Upon functional analyses during mouse male meiosis, we demonstrated that HELLS is required for PRDM9 binding and DSB activity at PRDM9 sites. However, HELLS is not required for DSB activity at PRDM9-independent sites. HELLS is also essential for 5-hydroxymethylcytosine (5hmC) enrichment at PRDM9 sites. Analyses of 5hmC in mice deficient for SPO11, which catalyzes DSB formation, and in PRDM9 methyltransferase deficient mice reveal that 5hmC is triggered at DSB-prone sites upon PRDM9 binding and histone modification, but independent of DSB activity. These findings highlight the complex regulation of the chromatin and epigenetic environments at PRDM9-specified hotspots.
Data availability
PRIDE partner repository with the dataset identifier PXD017337. NGS data have been deposited at GEO under series record GSE145768. Source data have been provided for Figure 2C-D and Figure 3G
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PRDM9 activity depends on HELLS and promotes local 5-hydroxymethylcytosine enrichmentNCBI Gene Expression Omnibus, GSE145768.
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Extensive sex differences at the initiation of meiotic recombinationNCBI Gene Expression Omnibus, GSE99921.
Article and author information
Author details
Funding
ERC (322788)
- Bernard de Massy
MSDAVenir (Gene-IGH)
- Bernard de Massy
Fondation Bettencourt Schueller
- Bernard de Massy
Fondation pour la Recherche Médicale
- Mathilde Biot
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All experiments were carried out according to the CNRS guidelines and were approved by the ethics committee on live animals (project CE-LR-0812 and 1295).
Copyright
© 2020, Imai et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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PRDM9 activity depends on HELLS and promotes local 5-hydroxymethylcytosine enrichment
eLife 9:e57117.
https://doi.org/10.7554/eLife.57117
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