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Transcriptional regulation of Wnt inhibitory factor-1 by Miz-1/c-Myc

Oncogene volume 29, pages 5923–5934 (2010)Cite this article

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Abstract

The Wnt signaling pathway is capable of self-regulation through positive and negative feedback mechanisms. For example, the oncoprotein c-Myc, which is upregulated by Wnt signaling activity, participates in a positive feedback loop of canonical Wnt signaling through repression of Wnt antagonists DKK1 and SFRP1. In this study, we investigated the mechanism of Wnt inhibitory factor-1 (WIF-1) silencing. Mapping of CpG island methylation of the WIF-1 promoter reveals regional methylation (−295 to −95 bp from the transcription start site) that correlates with transcriptional silencing. We identified Miz-1 as a direct activator of WIF-1 transcriptional activity, which is found at WIF-1 promoter. In addition, we show that c-Myc contributes to WIF-1 transcriptional repression in a Miz-1-dependent manner. Although the transient repression mediated by Miz-1/c-Myc is independent of de novo methylation, the stable repression by this complex is associated with CpG island methylation of the critical −295 to −95-bp region of the WIF-1 promoter. Importantly, Miz-1 and c-Myc are found at WIF-1 promoter in WIF-1 non-expressing cell lines DLD-1 and 209myc. Transient knockdown or somatic knockout of c-Myc in DLD-1 failed to restore WIF-1 expression suggesting that c-Myc is involved in initiating rather than maintaining WIF-1 epigenetic silencing. In a genome-wide screen, DNAJA4, TGFβ-induced and TRIM59 were repressed by c-Myc overexpression and DNA promoter hypermethylation. Our data reveal novel insights into c-Myc-mediated DNA methylation-dependent transcriptional silencing, a mechanism that might contribute to the dysregulation of Wnt signaling in cancer.

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Abbreviations

ChIP:

chromatin immunoprecipitation

MSP:

methylation-specific PCR

NSCLC:

non-small cell lung cancer

TSS:

transcription start site

WIF-1 :

Wnt inhibitory factor-1

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Acknowledgements

We acknowledge Dr Martin Eilers (Marbourg University, Germany), Dr Arthur Riggs (Beckman Research Institute, Duarte, CA, USA) and Dr Bert Vogelstein (Johns Hopkins School of Medicine, USA) for plasmids, Dr Kurt Bachman (University of Maryland, USA) for the DLD-1 c-Myc −/− DNA and cDNA. We are grateful to Dr Wayne Yu at the Johns Hopkins Microarray Core facility, Dr Kornel Schuebel for his help with the microarray experiment and Dr Mariann Bienz for her support. Finally, we thank all the members of the Baylin/Herman laboratory for useful discussions and Kathy Bender for administrative assistance. This study was supported by NCI/SPORE grant CA058184 to James G Herman, AICR grant 07-0040 to Mariann Bienz (MRC-LMB) and NCI grant P30 CA06973-44 to Leslie Cope.

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Authors and Affiliations

  1. Cancer Biology Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD, USA

    J D F Licchesi, V K Tiwari, L Cope, S B Baylin & J G Herman

  2. MRC Laboratory of Molecular Biology, Cambridge, UK

    J D F Licchesi

  3. Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium

    L Van Neste

  4. Department of Applied Mathematics and Statistics, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD, USA

    X Lin

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  1. J D F Licchesi
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Correspondence to J D F Licchesi or J G Herman.

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Competing interests

The commercial rights to the MSP technique belong to OncoMethylome Sciences. SBB and JGH serve as consultants to OncoMethylome Sciences and are entitled to royalties from any commercial use of this procedure. SBB and JGH also receive research support from OncoMethylome Sciences. The terms of this arrangement are being managed by the Johns Hopkins University in accordance with its conflict of interest policies.

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Licchesi, J., Van Neste, L., Tiwari, V. et al. Transcriptional regulation of Wnt inhibitory factor-1 by Miz-1/c-Myc. Oncogene 29, 5923–5934 (2010). https://doi.org/10.1038/onc.2010.322

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