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Posttranscriptional control of the replication stress response via TTP-mediated Claspin mRNA stabilization

Oncogene volume 39, pages 3245–3257 (2020)Cite this article

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Abstract

ATR and CHK1 play key roles in the protection and recovery of the stalled replication forks. Claspin, an adaptor for CHK1 activation, is essential for DNA damage signaling and efficient replication fork progression. Here, we show that tristetraprolin (TTP), an mRNA-binding protein, can modulate the replication stress response via stabilization of Claspin mRNA. TTP depletion compromised specifically in the phosphorylation of CHK1, but not p53 or H2AX among other ATR substrates, and produced CHK1-defective replication phenotypes including accumulation of stalled replication forks. Importantly, the expression of siRNA–resistant TTP in TTP-deficient cells restored CHK1 phosphorylation and reduced the number of stalled replication forks as close to the control cells. Besides, we found that TTP was required for efficient replication fork progression even in the absence of exogenous DNA damage in a Claspin-dependent manner. Mechanistically, TTP was able to bind to the 3′-untranslated region of Claspin mRNA to increase the stability of Claspin mRNA which eventually contributed to the subsequent ATR–CHK1 activation upon DNA damage. Taken together, our results revealed an intimate link between TTP-dependent Claspin mRNA stability and ATR–CHK1-dependent replication fork stability to maintain replication fork integrity and chromosomal stability.

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Fig. 1: Compromised CHK1 phosphorylation in response to genotoxic stress by TTP knockdown.
Fig. 2: TTP is required for the RSR.
Fig. 3: TTP stabilizes Claspin mRNA through a posttranscriptional mechanism.
Fig. 4: Restoration of the replication stress response by Claspin overexpression in TTP-deficient cells.
Fig. 5: TTP promotes normal replication fork velocity in a Claspin-dependent manner.
Fig. 6: TTP prevents genomic instability and apoptosis in response to replication stress.

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Acknowledgements

We would like to thank Dr Sang-Eun Lee (UT Health San Antonio) for the insightful discussion and comments. We also thank JWP (University of Ulsan) for generous sharing of reagents. This research was supported by Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education (NRF-2018R1D1A3B07043817 and NRF-2015R1D1A1A01056994).

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  1. These authors contributed equally: Tae-Hee Lee, Ji Ye Choi

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  1. Department of Biological Sciences, Dong-A University, Busan, 49315, South Korea

    Tae-Hee Lee, Ji Ye Choi, Jeong-Min Park & Tae-Hong Kang

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Lee, TH., Choi, J.Y., Park, JM. et al. Posttranscriptional control of the replication stress response via TTP-mediated Claspin mRNA stabilization. Oncogene 39, 3245–3257 (2020). https://doi.org/10.1038/s41388-020-1220-9

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