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The mTOR–S6K pathway links growth signalling to DNA damage response by targeting RNF168

Nature Cell Biology volume 20, pages 320–331 (2018)Cite this article

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Abstract

Growth signals, such as extracellular nutrients and growth factors, have substantial effects on genome integrity; however, the direct underlying link remains unclear. Here, we show that the mechanistic target of rapamycin (mTOR)–ribosomal S6 kinase (S6K) pathway, a central regulator of growth signalling, phosphorylates RNF168 at Ser60 to inhibit its E3 ligase activity, accelerate its proteolysis and impair its function in the DNA damage response, leading to accumulated unrepaired DNA and genome instability. Moreover, loss of the tumour suppressor liver kinase B1 (LKB1; also known as STK11) hyperactivates mTOR complex 1 (mTORC1)–S6K signalling and decreases RNF168 expression, resulting in defects in the DNA damage response. Expression of a phospho-deficient RNF168-S60A mutant rescues the DNA damage repair defects and suppresses tumorigenesis caused by Lkb1 loss. These results reveal an important function of mTORC1–S6K signalling in the DNA damage response and suggest a general mechanism that connects cell growth signalling to genome stability control.

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Fig. 1: mTORC1–S6K signalling suppresses the DDR.
Fig. 2: S6K phosphorylates RNF168 at Ser60.
Fig. 3: Ser60 phosphorylation inhibits RNF168 function and impairs DNA damage repair.
Fig. 4: RNF168 Ser60 phosphorylation promotes its degradation.
Fig. 5: Ser60 phosphorylation destabilizes RNF168 in a TRIP12-dependent manner.
Fig. 6: Inhibition of RNF168 by mTORC1–S6K contributes to DDR defects caused by Lkb1 loss.
Fig. 7: Phospho-deficient RNF168-SA mutant suppresses tumorigenesis in the KrasG12D/Lkb1L/L mice NSCLC model.

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Acknowledgements

We thank D. Durocher, C. Lucas and L. Penengo for sharing RNF168 constructs, and K.-L. Guan and D. Li for critical reading of the manuscript. This work was supported by the National Key Basic Research Program of China (2015CB964502), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000), and grants from the National Science foundation of China to D.G. (81372602 and 81422033) and to Xiaoduo Xie (31401214). P.L. is supported by 1K99CA181342 and 5T32HL007893 from National Institutes of Health (USA).

Author information

Author notes

  1. Pengda Liu

    Present address: Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

  2. Xiaoduo Xie, Hongli Hu, Xinyuan Tong and Long Li contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Cell Biology, CAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Innovation Center for Cell Signaling Network, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China

    Xiaoduo Xie, Hongli Hu, Xinyuan Tong, Long Li, Xiangyuan Liu, Min Chen, Yuxue Zhang, Huafang Ouyang, Hongbin Ji & Daming Gao

  2. The Key Laboratory of Stem Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, University of Chinese Academy of Sciences, Shanghai, China

    Huairui Yuan & Jun Qin

  3. State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China

    Xia Xie, Ronggui Hu & Fei-Long Meng

  4. CAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China

    Qingrun Li & Rong Zeng

  5. National Center for Protein Science Shanghai, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China

    Mengqi Wei & Jing Huang

  6. Shanghai Science Research Center, Chinese Academy of Sciences, Shanghai, China

    Mengqi Wei & Jing Huang

  7. Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Pengda Liu, Wenjian Gan & Wenyi Wei

  8. Department of Radiation Oncology, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, China

    Yong Liu

  9. Sir Run Run Shaw Hospital and Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

    Anyong Xie

  10. Department of Gastroenterology, Nantong University Affiliated Hospital, Nantong, Jiangsu, China

    Xiaoling Kuai

  11. OrigiMed Co. Ltd, Shanghai, China

    Gung-Wei Chirn

  12. Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China

    Hu Zhou

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Contributions

Xiaoduo Xie, H.H., X.T. and L.L. performed most of the experiments with assistance from X.L., M.C., Xia Xie, Y.Z., H.O., M.W., J.H., P.L., W.G., Y.L., A.X., X.K. and G.-W.C. H.Y. performed the villi length assay of mice after IR treatment under the supervision of J.Q. Q.L. and H.Z. performed the mass spectrometry analysis under the supervision of R.Z. D.G., Xiaoduo Xie, H.H.,W.W. and H.J. designed the experiments with input from R.H. F.-L.M., D.G., Xiaoduo Xie and H.H. wrote the manuscript. All authors commented on the manuscript.

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Correspondence to Daming Gao.

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Supplementary information

Supplementary Information

Supplementary Figures 1–8, Supplementary Table legends.

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Supplementary Table 1

Primary antibodies and reagents used in this study.

Supplementary Table 2

Target sequences of shRNAs and siRNAs used in this study.

Supplementary Table 3

Statistics source data.

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Xie, X., Hu, H., Tong, X. et al. The mTOR–S6K pathway links growth signalling to DNA damage response by targeting RNF168. Nat Cell Biol 20, 320–331 (2018). https://doi.org/10.1038/s41556-017-0033-8

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