Abstract
Deregulated expression of c-myc can induce cell proliferation in established cell lines and in primary mouse embryonic fibroblasts (MEFs), through a combination of both transcriptional activation and repression by Myc. Here we show that a Myc-associated transcription factor, Miz-1, arrests cells in G1 phase and inhibits cyclin D-associated kinase activity. Miz-1 upregulates expression of the cyclin-dependent kinases (CDK) inhibitor p15INK4b by binding to the initiator element of the p15INK4b promoter. Myc and Max form a complex with Miz-1 at the p15 initiator and inhibit transcriptional activation by Miz-1. Expression of Myc in primary cells inhibits the accumulation of p15INK4b that is associated with cellular senescence; conversely, deletion of c-myc in an established cell line activates p15INK4b expression. Alleles of c-myc that are unable to bind to Miz-1 fail to inhibit accumulation of p15INK4b messenger RNA in primary cells and are, as a consequence, deficient in immortalization.
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Acknowledgements
We thank P. Krimpenfort and A. Berns for p15+/+ and p15−/− fibroblasts, B. Eisenman for Mad-1 cDNA, several colleagues for anti-Max antibodies, K-H. Klempnauer for GST–p300 chimaeric proteins, R. Eckner for p300 clones and antibodies, D. Parry and Y. Xiong for anti-p15 antibodies, J. Sedivy for TGR and c-myc−/− cells, and K. Vousden for E7 plasmids. We also thank C. Pouponnot for help with bandshift experiments, and A. Maier, A. Grzeschiczek and A. A. Kjerulff for technical support. M.E. was supported by the Deutsche Forschungsgemeinschaft, the Biomed 2 programme of the European Community, and the Human Frontiers of Science Organisation.
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Staller, P., Peukert, K., Kiermaier, A. et al. Repression of p15INK4b expression by Myc through association with Miz-1. Nat Cell Biol 3, 392–399 (2001). https://doi.org/10.1038/35070076
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DOI: https://doi.org/10.1038/35070076
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