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MBD1 protects replication fork stability by recruiting PARP1 and controlling transcription-replication conflicts

Cancer Gene Therapy volume 31, pages 94–107 (2024)Cite this article

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Abstract

The replication-stress response is essential to ensure the faithful transmission of genetic information to daughter cells. Although several stress-resolution pathways have been identified to deal with replication stress, the precise regulatory mechanisms for replication fork stability are not fully understood. Our study identified Methyl-CpG Binding Domain 1 (MBD1) as essential for the maintaining genomic stability and protecting stalled replication forks in mammalian cells. Depletion of MBD1 increases DNA lesions and sensitivity to replication stress. Mechanistically, we found that loss of MBD1 leads to the dissociation of Poly(ADP-ribose) polymerase 1 (PARP1) from the replication fork, potentially accelerating fork progression and resulting in higher levels of transcription-replication conflicts (T-R conflicts). Using a proximity ligation assay combined with 5-ethynyl-2′-deoxyuridine, we revealed that the MBD1 and PARP1 proteins were recruited to stalled forks under hydroxyurea (HU) treatment. In addition, our study showed that the level of R-loops also increased in MBD1-delated cells. Without MBD1, stalled replication forks resulting from T-R conflicts were primarily degraded by the DNA2 nuclease. Our findings shed light on a new aspect of MBD1 in maintaining genome stability and providing insights into the mechanisms underlying replication stress response.

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Fig. 1: Genetic deletion of MBD1 results in genome instability.
Fig. 2: Loss of MBD1 increases the sensitivity of cells to replication stress.
Fig. 3: MBD1 maintains replication fork stability in a RAD51 independent manner.
Fig. 4: MBD1 interacts with PARP1 and the interaction is modulated by replication stress and PARylation.
Fig. 5: MBD1 regulates replication fork speed by promoting PARP1 association with replication fork.
Fig. 6: MBD1 participates in R-loop-associated transcription-replication conflicts.
Fig. 7: MBD1 is a potential target to improve the effectiveness of chemotherapy.

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Acknowledgements

We are grateful to members of the Wang laboratory for insightful discussions and technical assistance.

Funding

We thank the following for funding support: the National Key R&D Program of China (2022YFA1302800), the National Science Fund for Distinguished Young Scholars (82125031), and the National Natural Science Foundation of China (82230089).

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Authors and Affiliations

  1. Department of Radiation Medicine, School of Basic Medical Sciences, Peking University International Cancer Institute, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science Center, Beijing, 100191, China

    Guihui Yu, Yundong Xiong, Zhanzhan Xu, Lei Zhang, Xiao Albert Zhou, Chen Nie, Shiwei Li, Weibin Wang, Xiaoman Li & Jiadong Wang

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  1. Guihui Yu
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Contributions

YGH was responsible for designing and performing the experiments. YGH contributed to writing the original draft. LXM and XYD was responsible for bioinformatics analysis. ZL was responsible for part of the construction of plasmids and COIP assay. XZZ, ZX, NC, and WWB provided feedback on the report. LSW was responsible for purchasing consumables and reagents. YGH, LXM, and WJD reviewed and edited the manuscript.

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Correspondence to Weibin Wang, Xiaoman Li or Jiadong Wang.

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Yu, G., Xiong, Y., Xu, Z. et al. MBD1 protects replication fork stability by recruiting PARP1 and controlling transcription-replication conflicts. Cancer Gene Ther 31, 94–107 (2024). https://doi.org/10.1038/s41417-023-00685-0

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