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A novel WEE1 pathway for replication stress responses

Nature Plants volume 7, pages 209–218 (2021)Cite this article

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Abstract

DNA replication stress poses a severe threat to genome stability and is a hallmark of cancer as well as a target for cancer therapy. It is well known that the evolutionarily conserved protein kinase WEE1 regulates replication stress responses by directly phosphorylating and inhibiting the major cell cycle driver CDKs in many organisms. Here, we report a novel WEE1 pathway. We found that Arabidopsis WEE1 directly interacts with and phosphorylates the E3 ubiquitin ligase FBL17 that promotes the degradation of CDK inhibitors. The phosphorylated FBL17 is further polyubiquitinated and degraded, thereby leading to the accumulation of CDK inhibitors and the inhibition of CDKs. In strong support for this model, either loss of function of FBL17 or overexpression of CDK inhibitors suppresses the hypersensitivity of the wee1 mutant to replication stress. Intriguingly, human WEE1 also phosphorylates and destabilizes the FBL17 equivalent protein SKP2, indicating that this is a conserved mechanism. This study reveals that the WEE1-FBL17/SKP2-CKIs-CDKs axis is a molecular framework for replication stress responses, which may have clinical implications because the WEE1 inhibitor AZD1775 is currently in phase II clinical trial as an anticancer drug.

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Fig. 1: Loss of function of FBL17 suppresses the hypersensitivity of atr and wee1 to replication stress.
Fig. 2: FBL17 interacts with WEE1.
Fig. 3: WEE1 phosphorylates FBL17.
Fig. 4: WEE1-mediated FBL17 phosphorylation promotes FBL17 degradation.
Fig. 5: KRP7 functions downstream of FBL17 in the WEE1 pathway.
Fig. 6: Human WEE1A phosphorylates and negatively regulates SKP2.

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Data availability

The data that support the findings of this study are available from the corresponding author upon request. Source data are provided with this paper.

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Acknowledgements

We are grateful to H. G. Nam for providing the pFBL17:FBL17-GFP vector and L. De Veylder for critically revising the manuscript. This work was supported by the National Natural Science Foundation of China (grant nos. 31571253, 31771355 and 31970311); Fundamental Research Funds for the Central Universities (grant no. 2662019PY029); Thousand Talents Plan of China-Young Professionals Grant; and Huazhong Agricultural University Scientific & Technological Self-innovation Foundation (grant no. 2014RC004).

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Authors and Affiliations

  1. College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China

    Ting Pan, Qi Qin, Chubing Nong, Shan Gao, Lili Wang, Bingcheng Cai, Ming Zhang, Chong Wu, Hanchen Chen, Tong Li & Shunping Yan

  2. College of Informatics, Huazhong Agricultural University, Wuhan, Hubei, China

    Dan Xiong & Guoliang Li

  3. College of Life Sciences, Shanghai Normal University, Shanghai, China

    Shui Wang

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Contributions

T.P. and S.Y. designed the experiments. T.P., Q.Q., C.N., S.G., L.W., B.C., M.Z., C.W., H.C., T.L., D.X., G.L. and S.W. carried out the experiments. T.P. and S.Y. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Shunping Yan.

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Peer review information Nature Plants thanks Pascal Genschik, Jingsong Yuan and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Pan, T., Qin, Q., Nong, C. et al. A novel WEE1 pathway for replication stress responses. Nat. Plants 7, 209–218 (2021). https://doi.org/10.1038/s41477-021-00855-8

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