Abstract
Lethal (3) malignant brain tumor like 2 (L3MBTL2) is a member of the MBT-domain proteins, which are involved in transcriptional repression and implicated in chromatin compaction. Our previous study has shown that L3MBTL2 is highly expressed in the testis, but its role in spermatogenesis remains unclear. In the present study, we found that L3MBTL2 was most highly expressed in pachytene spermatocytes within the testis. Germ cell-specific ablation of L3mbtl2 in the testis led to increased abnormal spermatozoa, progressive decrease of sperm counts and premature testicular failure in mice. RNA-sequencing analysis on L3mbtl2 deficient testes confirmed that L3MBTL2 was a transcriptional repressor but failed to reveal any significant changes in spermatogenesis-associated genes. Interestingly, L3mbtl2 deficiency resulted in increased γH2AX deposition in the leptotene spermatocytes, subsequent inappropriate retention of γH2AX on autosomes, and defective crossing-over and synapsis during the pachytene stage of meiosis I, and more germ cell apoptosis and degeneration in aging mice. L3MBTL2 interacted with the histone ubiquitin ligase RNF8. Inhibition of L3MBTL2 reduced nuclear RNF8 and ubH2A levels in GC2 cells. L3mbtl2 deficiency led to decreases in the levels of the RNF8 and ubH2A pathway and in histone acetylation in elongating spermatids, and in protamine 1 deposition and chromatin condensation in sperm. These results suggest that L3MBTL2 plays important roles in chromatin remodeling during meiosis and spermiogenesis.
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Acknowledgements
We are grateful to Dr. P. Jeremy Wang (University of Pennsylvania) for helpful suggestion and critical reading and editing of the manuscript. We thank Josie Lai and Jean Kung (The Chinese University of Hong Kong) for technical assistance with the electronic microscopy. We also thank Dr. Wenhui Zhao (Peking University) for the CMV-HA-RNF8 plasmid, and Dr. Winnie Shum (ShanghaiTech University) for helpful discussions. Mr. Geng Tian (Shanghai Jiao Tong University) helped with the bioinformatics analysis. This work was supported by Health and Medical Research Fund (HMRF) grants 04153586 and 05161376 (to Y. X.).
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Meng, C., Liao, J., Zhao, D. et al. L3MBTL2 regulates chromatin remodeling during spermatogenesis. Cell Death Differ 26, 2194–2207 (2019). https://doi.org/10.1038/s41418-019-0283-z
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DOI: https://doi.org/10.1038/s41418-019-0283-z
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