Abstract

Cyclin C was cloned as a growth-promoting G1 cyclin, and was also shown to regulate gene transcription. Here we report that in vivo cyclin C acts as a haploinsufficient tumour suppressor, by controlling Notch1 oncogene levels. Cyclin C activates an ‘orphan’ CDK19 kinase, as well as CDK8 and CDK3. These cyclin-C–CDK complexes phosphorylate the Notch1 intracellular domain (ICN1) and promote ICN1 degradation. Genetic ablation of cyclin C blocks ICN1 phosphorylation in vivo, thereby elevating ICN1 levels in cyclin-C-knockout mice. Cyclin C ablation or heterozygosity collaborates with other oncogenic lesions and accelerates development of T-cell acute lymphoblastic leukaemia (T-ALL). Furthermore, the cyclin C encoding gene CCNC is heterozygously deleted in a significant fraction of human T-ALLs, and these tumours express reduced cyclin C levels. We also describe point mutations in human T-ALL that render cyclin-C–CDK unable to phosphorylate ICN1. Hence, tumour cells may develop different strategies to evade inhibition by cyclin C.

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Acknowledgements

We thank S. Blacklow, T. Sanda, T. Roberts, E. Sicinska, I. Kalaszczynska, S. Bukarac and D. Payne-Turner for help at different stages of the project. This work was supported by grants P01 CA109901 (to P.S., H.v.B. and A.T.L.), R01 CA083688 (to P.S.), GM094777 (to W.W.), American Syrian Lebanese Associated Charities of St. Jude Children’s Research Hospital (to C.G.M.), and R01 AG011085 (to J.W.H.). C.G.M. is a St. Baldrick’s Scholar and Pew Scholar.

Author information

Author notes

    • Anne Fassl
    • , Joel Chick
    • , Hiroyuki Inuzuka
    • , Xiaoyu Li
    •  & Marc R. Mansour

    These authors contributed equally to this work.

Affiliations

  1. Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Na Li
    • , Anne Fassl
    • , Lijun Liu
    • , Haizhen Wang
    • , Tobias Otto
    • , Lukas Baitsch
    • , Leah Bury
    • , Nan Ke
    • , Kristin A. Mulry
    • , Yan Geng
    • , Agnieszka Zagozdzon
    • , Jean J. Zhao
    •  & Piotr Sicinski
  2. Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Na Li
    • , Anne Fassl
    • , Lijun Liu
    • , Haizhen Wang
    • , Tobias Otto
    • , Leah Bury
    • , Nan Ke
    • , Kristin A. Mulry
    • , Yan Geng
    • , Agnieszka Zagozdzon
    •  & Piotr Sicinski
  3. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Joel Chick
    • , Alban Ordureau
    • , J. Wade Harper
    •  & Steven P. Gygi
  4. Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Hiroyuki Inuzuka
    • , Shavali Shaik
    •  & Wenyi Wei
  5. Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Xiaoyu Li
    • , Taras Kreslavsky
    •  & Harald von Boehmer
  6. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Xiaoyu Li
    • , Taras Kreslavsky
    •  & Harald von Boehmer
  7. Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Marc R. Mansour
    • , Alejandro Gutierrez
    •  & A. Thomas Look
  8. Division of Hematology/Oncology, Children’s Hospital, Boston, Massachusetts 02115, USA

    • Marc R. Mansour
    • , Alejandro Gutierrez
    •  & A. Thomas Look
  9. Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Marc R. Mansour
    • , Alejandro Gutierrez
    •  & A. Thomas Look
  10. Department of Haematology, University College London Cancer Institute, London WC1E 6BT, UK

    • Marc R. Mansour
    • , Sarah Jenkinson
    • , Rosemary E. Gale
    •  & David C. Linch
  11. Howard Hughes Medical Institute and Department of Pathology, NYU School of Medicine, New York, New York 10016, USA

    • Bryan King
    •  & Iannis Aifantis
  12. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Lukas Baitsch
    •  & Jean J. Zhao
  13. Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Clifford A. Meyer
  14. Harvard School of Public Health, Boston, Massachusetts 02115, USA

    • Clifford A. Meyer
  15. Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Michael J. Kluk
    • , Moni Roy
    •  & Jon C. Aster
  16. Novartis Institutes for BioMedical Research, Cambridge, Massachusetts 02139, USA

    • Sunkyu Kim
  17. Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, USA

    • Xiaowu Zhang
  18. Department of Pathology, St. Jude Research Hospital, Memphis, Tennessee 38105, USA

    • Charles G. Mullighan

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Contributions

N.L. and P.S. conceived the study, analysed and interpreted the data and wrote the paper. N.L. performed experiments described in the study with fundamental help from: A.F. who contributed phenotypic, in vivo molecular and tumour analyses, and contributed to writing the manuscript, J.C. and S.P.G. contributed all mass spectrometry analyses, H.I., S.S. and W.W. contributed molecular mechanistic analyses of the cyclin C → Fbw7 → ICN1 link, X.L. and H.v.B. contributed bone marrow transduction with ICN1 experiments, T.K. and H.v.B. performed analyses of thymocyte populations and studies of bone marrow chimaeras. M.R.M., S.J., R.E.G. and D.C.L. discovered mutations in human patients that render cyclin C–CDK unable to phosphorylate Notch1, L.L. contributed to molecular studies and phosphorylation analyses, H.W. performed all analyses of ICN1 phosphorylation by cyclin C–CDK8, C–CDK19 and C–CDK3, B.K. and I.A. contributed in vivo mouse tumorigenesis studies, A.G. and A.T.L. contributed analyses of human T-ALL samples, A.O. and J.W.H. performed analyses of endogenous ICN1 polyubiquitylation, T.O. contributed shRNA analyses of mouse cells and analyses of pRB phosphorylation, L. Baitsch and J.J.Z. helped with molecular in vivo analyses, C.A.M. analysed gene expression data, M.J.K. and J.C.A. performed immunostaining for ICN1, M.R. and J.C.A. contributed analyses of anti-phospho ICN1 antibody, S.K. helped with analyses of human T-ALL, X.Z. and J.C.A. developed the anti-phospho-ICN1 antibody, C.G.M. contributed DNA sequencing and copy number analyses of the cyclin C gene in human T-ALL, L. Bury, N.K., K.A.M., Y.G. and A.Z. helped N.L. at different stages of the project. A.F., J.C., H.I., X.L. and M.R.M. contributed equally. P.S. directed the study.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Piotr Sicinski.

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https://doi.org/10.1038/ncb3046

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