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UBE2T resolves transcription-replication conflicts and protects common fragile sites in primordial germ cells

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Abstract

The proper development of primordial germ cells (PGCs) is an essential prerequisite for gametogenesis and mammalian fertility. The Fanconi anemia (FA) pathway functions in maintaining the development of PGCs. FANCT/UBE2T serves as an E2 ubiquitin-conjugating enzyme that ubiquitylates the FANCD2-FANCI complex to activate the FA pathway, but its role in the development of PGCs is not clear. In this study, we found that Ube2t knockout mice showed defects in PGC proliferation, leading to severe loss of germ cells after birth. Deletion of UBE2T exacerbated DNA damage and triggered the activation of the p53 pathway. We further demonstrated that UBE2T counteracted transcription-replication conflicts by resolving R-loops and stabilizing replication forks, and also protected common fragile sites by resolving R-loops in large genes and promoting mitotic DNA synthesis to maintain the genome stability of PGCs. Overall, these results provide new insights into the function and regulatory mechanisms of the FA pathway ensuring normal development of PGCs.

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Data availability

The sequencing data are available in Gene Expression Omnibus (GEO) database with the accession number GSE223410 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE223410). Enter token ujexukgqhtiltor into the box to review the data.

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Acknowledgements

The authors thank Prof. Ping Zheng from Kunming Institute of Zoology, Chinese Academy of Sciences for providing technical support about DNA fiber assay and comet assay.

Funding

This work was supported by the National Key Research and Development Program of China [2022YFC2703800 and 2021YFC2700100]; Basic Science Center Program of NSFC [31988101]; National Natural Science Foundation for Distinguished Young Scholars [82125014]; National Natural Science Foundation of China [32170867 and 82071609]; Natural Science Foundation of Shandong Province for Grand Basic Projects [ZR2021ZD33]; Shandong Provincial Key Research and Development Program [2020ZLYS02]; Research Unit of Gametogenesis and Health of ART-Offspring; Chinese Academy of Medical Sciences [2020RU001]; Taishan Scholars Program for Young Experts of Shandong Province; and Qilu Young Scholars Program of Shandong University.

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Authors and Affiliations

  1. Center for Reproductive Medicine, Shandong University, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  2. Key Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  3. Shandong Key Laboratory of Reproductive Medicine, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  4. Shandong Provincial Clinical Research Center for Reproductive Health, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  5. Shandong Technology Innovation Center for Reproductive Health, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  6. National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, 250012, Shandong, China

    Yongze Yu, Weiwei Xu, Canxin Wen, Simin Zhao, Guangyu Li, Ran Liu, Zi-Jiang Chen, Yingying Qin, Jinlong Ma, Yajuan Yang & Shidou Zhao

  7. Research Unit of Gametogenesis and Health of ART-Offspring, Chinese Academy of Medical Sciences, Jinan, 250021, Shandong, China

    Zi-Jiang Chen

  8. Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China

    Zi-Jiang Chen

  9. Center for Reproductive Medicine, School of Medicine, Ren Ji Hospital, Shanghai Jiao Tong University, Shanghai, 200135, China

    Zi-Jiang Chen

Authors
  1. Yongze Yu
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  2. Weiwei Xu
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  3. Canxin Wen
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  4. Simin Zhao
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  5. Guangyu Li
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  6. Ran Liu
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  7. Zi-Jiang Chen
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  8. Yingying Qin
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  9. Jinlong Ma
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  10. Yajuan Yang
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  11. Shidou Zhao
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Contributions

Shidou Zhao, YQ, and JM designed the study. Y Yu performed most experiments; Y Yang completed the DNA fiber assay; WX completed the neutral comet assay; CW and Simin Zhao helped with the generation of MEFs; GL and RL participated in the genotyping; The manuscript was written by Y Yu and revised by Shidou Zhao, YQ and Z-JC.

Corresponding authors

Correspondence to Jinlong Ma, Yajuan Yang or Shidou Zhao.

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The authors declare no competing interests.

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All animal experiments were conducted in accordance with the ethical guidelines approved by the Animal Care and Research Committee of Shandong University.

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Yu, Y., Xu, W., Wen, C. et al. UBE2T resolves transcription-replication conflicts and protects common fragile sites in primordial germ cells. Cell. Mol. Life Sci. 80, 92 (2023). https://doi.org/10.1007/s00018-023-04733-8

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