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Melanocyte transformation requires complete loss of all pocket protein function via a mechanism that mitigates the need for MAPK pathway activation

Oncogene volume 36, pages 3789–3795 (2017)Cite this article

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Abstract

Deregulation of p16INK4A is a critical event in melanoma susceptibility and progression. It is generally assumed that the major effect of loss of p16 function is mediated through the CDK−cyclin pathway via its influence on the pocket protein (PP) pRb. However, there are also two other PPs, p107 and p130, which, when phosphorylated by CDK–cyclin complexes, play a role in permitting cell progression. Cohorts of mice carrying melanocyte-specific knockouts (KOs) of various combinations of the three PPs were generated. Mice null for pRb, p107, p130 or any combination of double mutants did not develop melanoma. Surprisingly, melanocyte-specific loss of all three PPs facilitated melanoma development (median age of onset 308 days, penetrance 40% at 1 year). Tumorigenesis was exacerbated by Trp53 co-deletion (median age of onset 275 days, penetrance 82% at 1 year), with cell culture studies indicating that this difference may result from the apoptotic role of Trp53. Melanomas in PP;Trp53-deficient mice lacked either Ras or Braf mutations, and hence developed in the absence of constitutive MAPK pathway activation. The lag period between induction of total PP or PP/Trp53 KO and melanoma development indicates that additional genetic or epigenetic alterations may account for neoplastic progression. However, exome sequencing of PP;Trp53 KO melanomas failed to reveal any additional recurrent driver mutations. Analysis of the putative mutation signature of the PP;Trp53 KO melanomas suggests that melanocytes are primed for transformation via a mutagenic mechanism involving an excess of T>G substitutions, but not involving a preponderance of C>T substitutions at CpG sites, which is the case for most spontaneous cancers not driven by a specific carcinogen. In sum, deregulation of all three PPs appears central to neoplastic progression for melanoma, and the customary reference to the p16INKA/CDK4/pRB pathway may no longer be accurate; all PPs are potentially critical targets of CDK-cyclins in melanoma.

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Acknowledgements

This work was supported by research grants and fellowships to GFK and GW from the Cancer Council of Queensland. NKH is supported by a National Health and Medical Research Council Fellowship. We are grateful to Linda Chin and Marcus Bosenberg for the gift of the Tyr::CreERT2 mice.

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Authors and Affiliations

  1. Cell and molecular Biology Department, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia

    I D Tonks, P Mukhopadhyay, W A Schroder, A W Mould, H Y Handoko, B Ferguson, P Keith, N K Hayward, G J Walker & G F Kay

  2. School of Anatomy, Physiology and Human Biology, The University of Western Australia, Crawley, Western Australia, Australia

    A Sorolla

  3. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, UK

    A W Mould

  4. Department of Pathology, University of Tasmania School of Medicine, Hobart, Tasmania, Australia

    H K Muller

  5. Diamantina Institute for Cancer, Immunology and Metabolic Medicine, University of Queensland, Brisbane, Australia

    P Keith

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Correspondence to G J Walker.

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Tonks, I., Mukhopadhyay, P., Schroder, W. et al. Melanocyte transformation requires complete loss of all pocket protein function via a mechanism that mitigates the need for MAPK pathway activation. Oncogene 36, 3789–3795 (2017). https://doi.org/10.1038/onc.2016.511

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