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Updated: Dec 22, 2025
Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
Published on: September 28, 2012
The histone modification reader ZCWPW1 links histone methylation to PRDM9-induced double-strand break repair
Tao Huang1,2,3,4, Shenli Yuan5,6, Lei Gao5
1Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, China.
Abstract:
The histone modification writer Prdm9 has been shown to deposit H3K4me3 and H3K36me3 at future double-strand break (DSB) sites during the very early stages of meiosis, but the reader of these marks remains unclear. Here, we demonstrate that Zcwpw1 is an H3K4me3 reader that is required for DSB repair and synapsis in mouse testes. We generated H3K4me3 reader-dead Zcwpw1 mutant mice and found that their spermatocytes were arrested at the pachytene-like stage, which phenocopies the Zcwpw1 knock-out mice. Based on various ChIP-seq and immunofluorescence analyses using several mutants, we found that Zcwpw1's occupancy on chromatin is strongly promoted by the histone-modification activity of PRDM9. Zcwpw1 localizes to DMC1-labelled hotspots in a largely Prdm9-dependent manner, where it facilitates completion of synapsis by mediating the DSB repair process. In sum, our study demonstrates the function of ZCWPW1 that acts as part of the selection system for epigenetics-based recombination hotspots in mammals.
Insights
The histone reader Zcwpw1 is essential for repairing DNA double-strand breaks (DSBs) and enabling chromosome pairing during meiosis. Its function, dependent on the histone writer Prdm9, is crucial for mammalian reproduction.
Area of Science:
- Epigenetics
- Molecular Biology
- Reproductive Biology
Background:
- Prdm9 deposits H3K4me3 and H3K36me3 marks at future double-strand break (DSB) sites during early meiosis.
- The specific protein that reads these histone marks to guide DSB repair and subsequent meiotic progression remained unidentified.
Purpose of the Study:
- To identify the reader of H3K4me3 marks deposited by Prdm9.
- To elucidate the function of this reader in meiosis, specifically in DSB repair and synapsis.
Main Methods:
- Generation and analysis of H3K4me3 reader-dead Zcwpw1 mutant mice.
- Chromatin immunoprecipitation sequencing (ChIP-seq) to assess protein occupancy.
- Immunofluorescence microscopy to visualize protein localization and meiotic progression.
Main Results:
- Zcwpw1 was identified as an H3K4me3 reader crucial for DSB repair and synapsis in mouse testes.
- Zcwpw1 mutant mice exhibited spermatocyte arrest at the pachytene-like stage, mirroring Zcwpw1 knockout phenotypes.
- Zcwpw1 chromatin occupancy is dependent on PRDM9's histone modification activity and localizes to PRDM9-dependent recombination hotspots.
Conclusions:
- Zcwpw1 functions as an H3K4me3 reader, mediating DSB repair and facilitating synapsis completion.
- Zcwpw1 acts in concert with PRDM9, forming part of the epigenetic machinery that selects mammalian recombination hotspots.
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