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ARF-induced downregulation of Mip130/LIN-9 protein levels mediates a positive feedback that leads to increased expression of p16Ink4a and p19Arf

Oncogene volume 29, pages 1976–1986 (2010)Cite this article

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Abstract

The ARF-MDM2-p53 pathway constitutes one of the most important mechanisms of surveillance against oncogenic transformation, and its inactivation occurs in a large proportion of cancers. Here, we show that ARF regulates Mip130/LIN-9 by inducing its translocation to the nucleolus and decreasing the expression of the Mip130/LIN-9 protein through a post-transcriptional mechanism. The knockdown of Mip130/LIN-9 in p53−/− and Arf−/− mouse embryonic fibroblasts (MEFs) mimics some effects of ARF, such as the downregulation of B-Myb, impaired induction of G2/M genes, and a decrease in cell proliferation. Importantly, although the knockdown of Mip130/LIN-9 reduced the proliferation of p53 or Arf-null MEFs, only p53−/− MEFs showed a senescence-like state and an increase in the expression of Arf and p16. Interestingly, the increase in p16 and ARF is indirect because the Mip130/LIN-9 knockdown decreased the transcription of negative regulators of the Ink4a/Arf locus, such as BUBR1 and CDC6. Chromatin immunoprecipitation assays also reveal that Mip130/LIN-9 occupies the promoters of the BubR1 and cdc6 genes, suggesting that Mip130/LIN-9 is necessary for the expression of these genes. Altogether, these results indicate that there is a feedback mechanism between ARF and Mip130/LIN-9 in which either the increase of ARF or the decrease in Mip130/LIN-9 causes a further increase in the expression of Arf and p16.

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Abbreviations

MEFs:

mouse embryonic fibroblasts

CDK:

cyclin-dependent kinase

Mip:

Myb-interacting protein

WT:

wild type

GFP:

green fluorescence protein

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Acknowledgements

The authors thank Drs Charles Sherr and Martine Roussel, St Jude Children's Research Hospital, for the Arf and p53 plasmid constructs and Arf−/− and p53−/− MEFs. This work was supported by Public Health Service Grants RO1 GM81562 (ORC) and NCI-KO1 CA127862 (RS) from the National Institutes of Health. JS and MP were supported by NIH Institutional T32 training grants, Ruth L Kirschstein National Research Service Award NOT-OD-06-093 and Training Program in Signal Transduction and Cellular Endocrinology DK07739, respectively.

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Authors and Affiliations

  1. Department of Pharmacology, University of Illinois, Chicago, IL, USA

    J Song, R Sandoval, M A Pilkinton & O R Colamonici

  2. Department of Physiology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA

    X Tian

  3. Department of Biochemistry/Molecular Genetics, University of Illinois, Chicago, IL, USA

    P Raychaudhuri

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  1. J Song
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  3. M A Pilkinton
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Correspondence to O R Colamonici.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Song, J., Sandoval, R., Pilkinton, M. et al. ARF-induced downregulation of Mip130/LIN-9 protein levels mediates a positive feedback that leads to increased expression of p16Ink4a and p19Arf. Oncogene 29, 1976–1986 (2010). https://doi.org/10.1038/onc.2009.485

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