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Novel role for Ddx39 in differentiation and telomere length regulation of embryonic stem cells

Cell Death & Differentiation (2024)Cite this article

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Abstract

Erk signaling is indispensable for the self-renewal and differentiation of mouse embryonic stem cells (ESCs), as well as telomere homeostasis. But how Erk regulates these biological processes remains unclear. We identified 132 Erk2 interacting proteins by co-immunoprecipitation and mass spectrometric analysis, and focused on Ddx39 as a potential Erk2 substrate. We demonstrated that Erk2 phosphorylates Ddx39 on Y132 and Y138. Ddx39 knockout (KO) ESCs are defective in differentiation, due to reduced H3K27ac level upon differentiation. Phosphorylation of Ddx39 promotes the recruitment of Hat1 to acetylate H3K27 and activate differentiation genes. In addition, Ddx39 KO leads to telomere elongation in ESCs. Ddx39 is recruited to telomeres by the telomere-binding protein Trf1, consequently disrupting the DNA loop formed by Trf1 and suppressing the alternative lengthening of telomeres (ALT). Phosphorylation of Ddx39 weakens its interaction with Trf1, releasing it from telomeres. Thus, ALT activity is enhanced, and telomeres are elongated. Altogether, our studies reveal an essential role of Ddx39 in the differentiation and telomere homeostasis of ESCs.

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Fig. 1: Erk2 phosphorylates Ddx39 at Y132 and Y138.
Fig. 2: Ddx39 depletion impairs differentiation capacity of ESCs.
Fig. 3: Ddx39 recruits Hat1 to activates differentiation genes.
Fig. 4: Ddx39 depletion leads to telomere elongation and enhanced ALT activity.
Fig. 5: Regulation of telomere homeostasis by Ddx39 is Trf1 dependent.
Fig. 6: Ddx39 mediates the regulatory effect of Erk on telomeres.

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Data availability

The raw sequencing data reported in this paper have been deposited in the GenBase in National Genomics Data Center (https://ngdc.cncb.ac.cn/) Beijing Institute of Genomics, Chinese Academy of Sciences/China National Center for Bioinformation, under the accession number: CRA009355. The mass spectrometry proteomics data have been deposited to the Proteome Xchange Consortium via the PRIDE partner repository with the dataset identifier PXD048543. The original western blot data are provided in Supplementary Materials (Original western blots).

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Acknowledgements

This work was supported by the National Key R&D Program of China [Grant No. 2021YFA1101002] and the 111 Project Grant [B08011].

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Authors and Affiliations

  1. State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Protein Sciences, Frontiers Science Center for Cell Responses, National Demonstration Center for Experimental Biology Education and College of Life Sciences, Nankai University, Tianjin, China

    Shanshan Nai, Jiao Yang, Bin Ling, Qiman Dong, Xiaoqiong Yang, Xiaoling Du, Lin Liu & Lingyi Chen

  2. State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University, Tianjin, China

    Meijie Wang, Man Lu & Zhongbo Yu

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SN, MW, JY, BL, QD, XY, ML and XD performed experiments, SN, LL and ZY analyzed the data and contributed to the paper writing, LC designed the experiments and wrote the paper.

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Correspondence to Lingyi Chen.

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Nai, S., Wang, M., Yang, J. et al. Novel role for Ddx39 in differentiation and telomere length regulation of embryonic stem cells. Cell Death Differ (2024). https://doi.org/10.1038/s41418-024-01354-x

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