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A chemical biology approach identifies AMPK as a modulator of melanoma oncogene MITF

Oncogene volume 33, pages 2531–2539 (2014)Cite this article

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Abstract

The microphthalmia-associated transcription factor (MITF) is indispensable for the viability of melanocytic cells, is an oncogene in melanoma and has a cell type-specific expression pattern. As the modulation of MITF activity by direct chemical targeting remains a challenge, we assessed a panel of drugs for their ability to downregulate MITF expression or activity by targeting its upstream modulators. We found that the multi-kinase inhibitors midostaurin and sunitinib downregulate MITF protein levels. To identify the target molecules shared by both the drugs in melanocytic cells, a chemical proteomic approach was applied and AMP-activated kinase (AMPK) was identified as the relevant target for the observed phenotype. RNA interference and chemical inhibition of AMPK led to a decrease in MITF protein levels. Reduction of MITF protein levels was the result of proteasomal degradation, which was preceded by enhanced phosphorylation of MITF mediated by ERK. As expected, downregulation of MITF protein levels by AMPK inhibition was associated with decreased viability. Together, these results identify AMPK as an important regulator for the maintenance of MITF protein levels in melanocytic cells.

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Acknowledgements

We thank Hans Widlund (Harvard Skin Disease Research Center, Brigham and Women’s Hospital, Boston, MA, USA) for providing us with human immortalised melanocytes with and without ectopic expression of BRAFV600E and HA-MITF as described in Garraway et al.1 We thank Gaurav Pathria for helpful discussions and critical reading of the manuscript. This study was funded by FWF-Austrian Science Fund (L590-B12) to SNW. The team at CeMM is supported by the Austrian Academy of Sciences, and the GEN-AU initiative of the Austrian Federal Ministry for Science and Research (PLACEBO GZ BMWF-70.081/0018-II/1a/2008 and APP-III 820965).

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  1. U Rix

    Present address: 3Current address: Drug Discovery Department, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA.,

Authors and Affiliations

  1. Division of Immunology, Department of Dermatology, Allergy and Infectious Diseases, Medical University of Vienna, Vienna, Austria

    V Borgdorff, C Wagner & S N Wagner

  2. CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria

    V Borgdorff, U Rix, G E Winter, M Gridling, A C Müller, F P Breitwieser, J Colinge, K L Bennett, G Superti-Furga & S N Wagner

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Borgdorff, V., Rix, U., Winter, G. et al. A chemical biology approach identifies AMPK as a modulator of melanoma oncogene MITF. Oncogene 33, 2531–2539 (2014). https://doi.org/10.1038/onc.2013.185

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