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Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway

Nature Cell Biology volume 8, pages 1053–1063 (2006)Cite this article

A Corrigendum to this article was published on 01 November 2006

Abstract

Dysfunction of the endoplasmic reticulum (ER) has been reported in a variety of human pathologies, including cancer. However, the contribution of the ER to the early stages of normal cell transformation is largely unknown. Using primary human melanocytes and biopsies of human naevi (moles), we show that the extent of ER stress induced by cellular oncogenes may define the mechanism of activation of premature senescence. Specifically, we found that oncogenic forms of HRAS (HRASG12V) but not its downstream target BRAF (BRAFV600E), engaged a rapid cell-cycle arrest that was associated with massive vacuolization and expansion of the ER. However, neither p53, p16INK4a nor classical senescence markers – such as foci of heterochromatin or DNA damage – were able to account for the specific response of melanocytes to HRASG12V. Instead, HRASG12V-driven senescence was mediated by the ER-associated unfolded protein response (UPR). The impact of HRAS on the UPR was selective, as it was poorly induced by activated NRAS (more frequently mutated in melanoma than HRAS). These results argue against premature senescence as a converging mechanism of response to activating oncogenes and support a direct role of the ER as a gatekeeper of tumour control.

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Figure 1: Differential senescence programmes induced by HRASV12 and BRAFE600 in primary melanocytes.
Figure 2: Classical markers of senescence are not essential for HRASV12-driven melanocyte senescence.
Figure 3: Expansion of the ER in HRASV12-senescent melanocytes.
Figure 4: Specific activation of the unfolded protein response by HRASV12.
Figure 5: Comparative analysis of the impact of HRASG12V and NRASQ61R in normal human melanocytes.
Figure 6: Analysis of ER stress markers in human naevi.

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Acknowledgements

We are grateful to A. Bengtson, T.P. Miller and W.-H. Tang for technical support; K. Zhang for advice with UPR-related dominant-negative constructs; J. Martens for help with the time-lapse microscopy; S. Núñez, E. de Stanchina and S. Lowe for p16 shRNA lentiviral vector and AKT-DN retroviral constructs; K. Guan and A. Vojtek for HA-tagged BRAF pcDNA; D. Peeper for pBabe-puro-BRAF; and W. Tirasophon and T. Ruthowski for ATF-6 and IRE1 antibodies. We also thank G. Núñez, J.A. Esteban, M. Serrano, G. Ferbeyre and A. Dlugosz for helpful suggestions and critical reading of this manuscript. This work was supported by a postdoctoral fellowship from La Ligue Contre le Cancer (C.D); Career Development Awards from the Dermatology Foundation to M.S.S. and M.A.N.; the Elsa U. Pardee Foundation (M.S.S); and National Institutes of Health grants CA107237 (M.S.S), R33 CA95300 (B.C.B.), DK42394 (R.J.K) and A112243 and U01CA83180 (S.B.G). M.S.S. is a V Foundation for Cancer Research Scholar.

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  1. Christophe Denoyelle, George Abou-Rjaily and Vladimir Bezrookove: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Dermatology and Comprehensive Cancer Center, University of Michigan, 1500E Medical Center Drive, 4217 CCGC, Ann Arbor, 48109, MI, USA

    Christophe Denoyelle, George Abou-Rjaily, Monique Verhaegen, Timothy M. Johnson, Mikhail A. Nikiforov & Maria S. Soengas

  2. Departments of Dermatology and Pathology and Comprehensive Cancer Center, University of California San Francisco, UCSF Box 0808, San Francisco, 94143, CA, USA

    Vladimir Bezrookove & Boris C. Bastian

  3. Department of Pathology, University of Michigan, 1500E Medical Center Drive, M5226 MSI, Ann Arbor, 48109, MI, USA

    Douglas R. Fullen & Lyndon D. Su

  4. Departments of Internal Medicine, Human Genetics and Epidemiology, University of Michigan, 1500E Medical Center Drive, 4301 MSRB III, Ann Arbor, 48109, MI, USA

    Jenny N. Pointer & Stephen B. Gruber

  5. Department of Biological Chemistry, University of Michigan and Howard Hughes Medical Institute, 4554 MSRB II, Ann Arbor, 48109, MI, USA

    Randal J. Kaufman

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Contributions

C.D. and M.S.S. designed the study. C.D., G.A-R., M.V. and M.S.S. performed all studies with primary melanocytes. T.M.J., D.R.F., J.P., S.B.G. and L.S. provided specimens for inital testing of stress factors in human naevi. Common and Spitz naevi shown or mentioned in this study were analysed for HRAS, BRAF and NRAS status by V.B. and B.C.B. V.B. and B.C.B. performed and scored the immunostaining of naevi with antibodies against PDI and GRP78, and M.S.S. performed statistical tests. M.A.N. and R.J.K. provided reagents for the generation of lentiviral vectors expressing shRNA against UPR proteins. C.D. and M.S.S. wrote the manuscript.

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Correspondence to Maria S. Soengas.

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Denoyelle, C., Abou-Rjaily, G., Bezrookove, V. et al. Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway. Nat Cell Biol 8, 1053–1063 (2006). https://doi.org/10.1038/ncb1471

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